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Clinical and Experimental Pharmacology and Physiology

Impact factor: 2.16 5-Year impact factor: 2.01 Print ISSN: 0305-1870 Online ISSN: 1440-1681 Publisher: Wiley Blackwell (Blackwell Publishing)

Subject: Psychology

Most recent papers:

  • Oncogenic role of neurotensin and neurotensin receptors in various cancers.
    Qing Ouyang, Ji Zhou, Wei Yang, Hongjuan Cui, Minhui Xu, Liang Yi.
    Clinical and Experimental Pharmacology and Physiology. 7 days ago
    Neurotensin (NTS) has long been recognized as a neurotransmitter or neuromodulator in the central nervous system and as an endocrine agent in the periphery via actions mediated through neurotensin receptors (NTSRs). Many studies support a role for NTS in the endocrine, autocrine and paracrine growth stimulation of cancer, with oncogenic actions described for NTS in different types of cancers and cancer cell lines at each step of cancer progression, ranging from tumour growth and survival to metastatic spread. The mechanisms underlying the effects of the NTS/NTSR system in cell proliferation, migration and invasion, as well as the anti‐apoptotic effects of this system, have been elucidated in different types of cancers, and include mitogen‐activated protein kinases, phosphatidylinositol 3‐kinase and RhoGTPases. The present mini review summarizes recent findings relating to the oncogenic function of the NTS/NTSR system.
    July 17, 2017   doi: 10.1111/1440-1681.12787   open full text
  • Naringenin prevents ischaemic stroke damage via anti‐apoptotic and anti‐oxidant effects.
    Kaihua Wang, Zhenzhen Chen, Jianmin Huang, Longjian Huang, Ning Luo, Xiulin Liang, Mingkun Liang, Wei Xie.
    Clinical and Experimental Pharmacology and Physiology. 7 days ago
    Apoptosis and oxidative stress are considered to be the major factors associated with the development and progression of many ischaemic cerebrovascular diseases. Naringenin (NAR) is an abundant flavanone in citrus plants and has been found to exhibit anti‐oxidant, anti‐carcinogenic and anti‐apoptotic effects. This study aimed to investigate the anti‐apoptotic and anti‐oxidant effects of naringenin on ischaemic stroke. In vitro, cortical neuron cells isolated from the brains of neonatal Sprague–Dawley rats were randomly divided into control, oxygen and glucose deprivation/reperfusion (OGD/Rep), NAR‐L, NAR‐M and NAR‐H groups. MTT and RT‐PCR were used for cell proliferation and apoptosis‐related proteins analyses. The effects of NAR on the Nrf2 signalling pathway were investigated using transfection approaches. Differences in mitochondrial dysfunction were analyzed by flow cytometry. In vivo, middle cerebral artery occlusion (MCAO) model was prepared and neurological defects and the brain wet/dry (W/D) ratio were assessed and recorded; apoptosis was measured based on the TUNEL assay. Additionally, biochemical indices were detected both in vitro and in vivo. NAR promoted cortical neuron cell proliferation, inhibited apoptosis and oxidative stress, and regulated the localization of Nrf2 protein (P<.05). Furthermore, silencing and overexpression of Nrf2 affected cortical neuron cell proliferation and apoptosis (P<.05). In vivo, NAR could alleviate cerebral oedema, improve neurological defects, and reduce apoptosis and oxidative stress (P<.05). These findings demonstrated that NAR could reduce apoptosis and oxidative stress and that Nrf2 signalling pathway is involved in this regulatory process. NAR has health‐promoting properties because of its anti‐apoptotic and anti‐oxidant effects in cases of ischaemic stroke.
    July 17, 2017   doi: 10.1111/1440-1681.12775   open full text
  • Proteomic analysis of oral cancer reveals new potential therapeutic targets involved in the Warburg effect.
    Yi‐Ping Huang, Nai‐Wen Chang.
    Clinical and Experimental Pharmacology and Physiology. 7 days ago
    Activation of peroxisome proliferator‐activated receptor alpha (PPARα) has been reported to disrupt tumour metabolism and to promote anticancer activity through interfering with the Warburg effect. This study is to investigate whether Warburg effect‐related proteins also could be identified in oral tumour lesions and to explore the functional significance of PPARα in metabolic shift. Five pairs of tongue tumour tissues and adjacent reference tissues obtained from 4‐NQO/arecoline induced mouse model were analyzed by 2‐d‐gel‐electrophoresis and LC‐MS. Further, the hexokinase II level, pyruvate dehydrogenase (PDH) activity, and metabolites of glycolysis and TCA cycle were all examined in order to validate the effect of PPARα on metabolic shift. Changes in protein expression levels revealed that seven proteins, which were involved in glycolysis, the tricarboxylic acid cycle, and the respiratory chain, were down‐regulated in tumour tissues. We found that activation of PPARα through fenofibrate could inhibit oral cancer cell growth and switch the way of energy production from the Warburg effect to oxidative phosphorylation. Fenofibrate induced a reduction of hexokinase II protein levels, increases in PDH activity and metabolites of the TCA cycle, and an impairment of ATP production. These findings suggested that activation of the PPARα to reprogram the metabolic pathway might impair the Warburg effect and trigger cancer cell death. The study provides a novel view of changes in protein expression profiles involved in the Warburg effect during oral tumourigenesis. Activation of the PPARα to impair the Warburg effect might offer a new strategy for oral cancer treatment.
    July 17, 2017   doi: 10.1111/1440-1681.12774   open full text
  • Highly expressed long non‐coding RNA CRNDE promotes cell proliferation through PI3K/AKT signalling in non‐small cell lung carcinoma.
    Xiao‐Xiong Liu, Han‐Peng Xiong, Jiu‐Sheng Huang, Kai Qi, Jian‐Jun Xu.
    Clinical and Experimental Pharmacology and Physiology. 7 days ago
    Recently, numerous studies have revealed that long non‐coding RNAs (lncRNAs) play complex roles in various lung diseases, while the colorectal neoplasia differentially expressed (CRNDE) functions in non‐small cell lung carcinomas (NSCLC) remain largely unknown. In the present study, we investigate the role and mechanism of CRNDE in the progression of NSCLC. The mRNA level of CRNDE in NSCLC patients and cells was detected by qRT‐PCR. The influence of CRNDE silencing or over‐expression on NSCLC cell proliferation and growth were assessed by MTT and flow cytometry, respectively. We also investigated the effect of abnormal CRNDE expression on cyclins and PI3K/AKT pathway. Furthermore, si‐CRNDE NSCLC cell lines were injected subcutaneously into nude mice to explore tumour formation in vivo. The expression of CRNDE was significantly upregulated in NSCLC patients and cells. In addition, both loss and gain function assays revealed that CRNDE promoted NSCLC cell proliferation and growth both in vitro and in vivo. Moreover, CRNDE regulated the cell cycle transition from G0/G1 stage to S stage and modulated the expression of CDK4, CDK6 and CCNE1. We further illustrated that CRNDE activated PI3K/AKT signalling in NSCLC cell lines. In conclusion, CRNDE was highly expressed in NSCLC malignant tissues and the heightened CRNDE strongly promoted NSCLC cell proliferation and growth through activating PI3K/AKT signalling; our results shed a light on utilizing CRNDE as a potential novel therapeutic target for the treatment of NSCLC.
    July 17, 2017   doi: 10.1111/1440-1681.12780   open full text
  • The opposite effects of nitric oxide donor, S‐nitrosoglutathione, on myocardial ischaemia/reperfusion injury in diabetic and non‐diabetic mice.
    Yi Liu, Chenhai Xia, Rutao Wang, Jinglong Zhang, Tao Yin, Yanzuo Ma, Ling Tao.
    Clinical and Experimental Pharmacology and Physiology. 7 days ago
    Nitric oxide is a potent anti‐apoptotic and cardioprotective molecule in healthy animals. However, recent study demonstrates that overexpression of eNOS exacerbates the liver injury in diabetic animals. whether diabetes may also alter NO's biologic activity in ischaemic/reperfused heart remains unknown. The present experiment was designed to determine whether the nitric oxide donor, S‐nitrosoglutathione, may exert different effects on diabetic and non‐diabetic myocardial ischaemia/reperfusion (MI/R) injury. Diabetic state was induced in mice by multiple intraperitoneal injections of low‐dose streptozotocin (STZ). The control or diabetic mice were subjected to 30 minutes ischaemia and 3 or 24 hours reperfusion. At 10 minutes before reperfusion, diabetic and non‐diabetic mice were received an intraperitoneal injection of S‐nitrosoglutathione (GSNO, a nitric oxide donor, 1 μmol/kg). GSNO attenuated MI/R injury in non‐diabetic mice, as measured by improved cardiac function, reduced infarct size and decreased cardiomyocyte apoptosis. In contrast, GSNO failed to attenuate but, rather, aggravated the MI/R injury in diabetic mice. Mechanically, the diabetic heart exhibited an increased nitrative/oxidative stress level, as measured by peroxynitrite formation, compared with non‐diabetic mice. Co‐administration of GSNO with EUK134 (a peroxynitrite scavenger) or MnTE‐2‐PyP5 (a superoxide dismutase mimetic) or Apocynin (a NADPH oxidase inhibitor) 10 minutes before reperfusion significantly decreased the MI/R‐induced peroxynitrite formation and the MI/R injury. Collectively, the present study for the first time demonstrated that diabetes may cause superoxide overproduction, increase NO inactivation and peroxynitrite formation, and thus convert GSNO from a cardioprotective molecule to a cardiotoxic molecule.
    July 17, 2017   doi: 10.1111/1440-1681.12781   open full text
  • Reduced platelet aggregation in women after intercourse: a possible role for the cyclooxygenase pathway.
    Gabriel Mayoral‐Andrade, Laura Pérez‐Campos‐Mayoral, Abraham Majluf‐Cruz, Eduardo Perez‐Campos Mayoral, Carlos Perez campos Mayoral, Ana Rocha‐Núñez, Margarito Martinez, Edgar Zenteno, Leticia Hernandez‐Gonzalez, María Guadalupe López Juan, Alma Dolores Pérez‐Santiago, Eduardo Pérez‐Campos.
    Clinical and Experimental Pharmacology and Physiology. 7 days ago
    We hypothesise that molecules in the cyclooxygenase pathway affect platelet activity when seminal fluid (SF) is present. We considered the influence of SF on platelet aggregation in women, and believe that the prostanoids in SF signalling are significant. Thirty‐one female subjects were studied, 20 of whom were sexually active. Male partners were given either aspirin or indomethacin to inhibit cyclooxygenase. The 6‐keto prostaglandin F1α (6‐keto PGF1α) and prostaglandin E metabolite (PGE‐M) in SF were measured by competitive assay. Platelets and prostanoids were evaluated in women, periodically, before and after intercourse. The platelets were tested with adenosine diphosphate (ADP) and arachidonic acid (AA). To block the interaction between the uterus and SF, some couples used condoms. We found that the 6‐keto prostaglandin F1α in urine at 2 hours post‐intercourse (1418.75 pg/mL, Std 688.39) was greater than pre‐intercourse (772.68 pg/mL, Std 116.54). Post‐intercourse, a transient decrease in platelet aggregation was observed in women whose partners did not use condoms. Averages for platelet aggregation were 20.16% with ADP, and more significantly, 37.79% with AA after 2 hours. In contrast, couples using condoms showed no changes, averaging 64.02% with ADP and 72.06% with AA. Women whose partners were taking aspirin or indomethacin also showed no changes. SF from men taking aspirin or indomethacin led to no reduction in platelet aggregometry in their partners. These results indicate that in cases of exposure to SF, the transient change in women's platelet activity could be related to the cyclooxygenase pathway.
    July 17, 2017   doi: 10.1111/1440-1681.12783   open full text
  • RNAi mediated gene silencing of ITPA using a targeted nanocarrier: Apoptosis induction in SKBR3 cancer cells.
    Fahimeh Charbgoo, Mehrdad Behmanesh, Maryam Nikkhah, Eric G Kane.
    Clinical and Experimental Pharmacology and Physiology. 7 days ago
    A pure nucleotide pool is required for high‐fidelity DNA replication and prevention of carcinogenesis in living cells. Human inosine triphosphatase (ITPase), encoded by the ITPA gene, plays a critical role in maintaining the purity of the cellular nucleotide pool by excluding nucleotides that enhance mutagenesis. ITPase is a nucleoside triphosphate pyrophosphatase that hydrolyzes the non‐canonical nucleotides inosine triphosphate (ITP) and xanthine triphosphate (XTP). The monophosphate products of ITPase reactions are subsequently excluded from the nucleotide pool and the improper substitution of ITP and XTP into DNA and RNA is prevented. Previous studies show that deficiency in ITPA can suppress cellular growth and enhance DNA instability. In this study, we evaluated the influence of effective ITPA down‐regulation on the induction of apoptosis in a human cancer cell line using folate‐single wall nanotubes (SWNT) as a targeted nanocarrier. We assessed whether SWNT enhances IPTA‐siRNA transfection efficiency in cancer cells using folate as a homing device. Since folate receptor is considerably overexpressed in cancer cells, conjugation of SWNTs to folate could enhance their cancer‐specific penetrance. We found that nanocarrier mediated ITPA‐siRNA transfection into SKBR3 cells caused significant reduction of ITPA mRNA expression level and complete down‐regulation of the ITPase protein product. The silencing of ITPA led to promotion of apoptosis in SWNT‐treated SKBR3 cancer cells.
    July 17, 2017   doi: 10.1111/1440-1681.12776   open full text
  • Subacute pyridostigmine exposure increases heart rate recovery and cardiac parasympathetic tone in rats.
    Manushree Bharadwaj, Carey Pope, Michael Davis, Stuart Katz, Christian Cook, Lara Maxwell.
    Clinical and Experimental Pharmacology and Physiology. 7 days ago
    Heart rate recovery (HRR) describes the rapid deceleration of heart rate after strenuous exercise and is an indicator of parasympathetic tone. A reduction in parasympathetic tone occurs in patients with congestive heart failure, resulting in prolonged HRR. Acetylcholinesterase inhibitors, such as pyridostigmine, can enhance parasympathetic tone by increasing cholinergic input to the heart. The objective of this study was to develop a rodent model of HRR to test the hypothesis that subacute pyridostigmine administration decreases cholinesterase activity and accelerates HRR in rats. Ten days after implantation of radiotelemetry transmitters, male Sprague Dawley rats were randomized to control (CTL) or treated (PYR; 0.14 mg/mL pyridostigmine in the drinking water, 29 days) groups. Rats were exercised on a treadmill to record HRR, and blood samples were collected on days 0, 7, 14, and 28 of pyridostigmine administration. Total cholinesterase and acetylcholinesterase (AChE) activity in plasma was decreased by 32%‐43% and 57%‐80%, respectively, in PYR rats on days 7‐28, while plasma butyrylcholinesterase activity did not significantly change. AChE activity in red blood cells was markedly reduced by 64%‐66%. HRR recorded 1 minute after exercise was higher in the PYR group on days 7, 14 and 28, and on day 7 when HRR was estimated at 3 and 5 minutes. Autonomic tone was evaluated pharmacologically using sequential administration of muscarinic (atropine) and adrenergic (propranolol) blockers. Parasympathetic tone was increased in PYR rats as compared with the CTL group. These data support the study hypothesis that subacute pyridostigmine administration enhances HRR by increasing cardiac parasympathetic tone.
    July 17, 2017   doi: 10.1111/1440-1681.12773   open full text
  • Angiotensin II‐preconditioning is associated with increased PKCε/PKCδ ratio and prosurvival kinases in mitochondria.
    Rebeca E. Nuñez, Sabzali Javadov, Nelson Escobales.
    Clinical and Experimental Pharmacology and Physiology. 10 days ago
    Angiotensin II preconditioning (APC) has been shown to reproduce the cardioprotective effects of ischaemic preconditioning (IPC), however, molecular mechanisms mediating the effects of APC remain unknown. In this study, Langendorff‐perfused rat hearts were subjected to IPC, APC or both (IPC/APC) followed by ischaemia‐reperfusion (IR), to determine translocation of PKCε, PKCδ, Akt, Erk1/2, JNK, p38 MAPK and GSK‐3β to mitochondria as an indicator of activation of the protein kinases. In agreement with previous observations, IPC, APC and IPC/APC increased the recovery of left ventricular developed pressure (LVDP), reduced infarct size (IS) and lactate dehydrogenase (LDH) release, compared to controls. These effects were associated with increased mitochondrial PKCε/PKCδ ratio, Akt, Erk1/2, JNK, and inhibition of permeability transition pore (mPTP) opening. Chelerythrine, a pan‐PKC inhibitor, abolished the enhancements of PKCε but increased PKCδ expression, and inhibited Akt, Erk1/2, and JNK protein levels. The drug had no effect on the APC‐ and IPC/APC‐induced cardioprotection as previously reported, but enhanced the post‐ischemic LVDP in controls. Losartan, an angiotensin II type 1 receptor (AT1‐R) blocker, abolished the APC‐stimulated increase of LVDP and reduced PKCε, Akt, Erk1/2, JNK, and p38. Both drugs reduced ischaemic contracture and LDH release, and abolished the inhibition of mPTP by the preconditioning. Chelerythrine also prevented the reduction of IS by APC and IPC/APC. These results suggest that the cardioprotection induced by APC and IPC/APC involves an AT1‐R‐dependent translocation of PKCε and survival kinases to the mitochondria leading to mPTP inhibition. In chelerythrine‐treated hearts, however, alternate mechanisms appear to maintain cardiac function. This article is protected by copyright. All rights reserved.
    July 14, 2017   doi: 10.1111/1440-1681.12816   open full text
  • Investigation of morin‐induced insulin secretion in cultured pancreatic cells.
    Mang Hung Lin, Chia‐Chen Hsu, Jenshinn Lin, Juei Tang Cheng, Ming Chang Wu.
    Clinical and Experimental Pharmacology and Physiology. 12 days ago
    Morin is a flavonoid contained in guava that is known to reduce hyperglycemia in diabetes. Insulin secretion has been demonstrated to increase following the administration of morin. The present study is designed to investigate the potential mechanism(s) of morin‐induced insulin secretion in the MIN6 cell line. First, we identified that morin induced a dose‐dependent increase in insulin secretion and intracellular calcium content in MIN6 cells. Morin potentiated glucose‐stimulated insulin secretion (GSIS). Additionally, we used siRNA for the ablation of imidazoline receptor protein (NISCH) expression in MIN6 cells. Interestingly, the effects of increased insulin secretion by morin and canavanine were markedly reduced in Si‐NISCH cells. Moreover, we used KU14R to block imidazoline I3 receptor (I‐3R) that is known to enhance insulin release from the pancreatic β‐cells. Without influence on the basal insulin secretion, KU14R dose‐dependently inhibited the increased insulin secretion induced by morin or efaroxan in MIN6 cells. Additionally, effects of increased insulin secretion by morin or efaroxan were reduced by diazoxide at the dose sufficient to open KATP‐channels and attenuated by nifedipine at the dose used to inhibit L‐type calcium channels. Otherwise, phospholipase C (PLC) is introduced to couple with imidazoline receptor (I‐R). The PLC inhibitor dose‐dependently inhibited the effects of morin in MIN6 cells. Similar blockade was also observed in protein kinase C (PKC) inhibitor‐treated cells. Taken together, we found that morin increases insulin secretion via the activation of I‐R in pancreatic cells. Therefore, morin would be useful to develop in the research and treatment of diabetic disorders. This article is protected by copyright. All rights reserved.
    July 12, 2017   doi: 10.1111/1440-1681.12815   open full text
  • (Pro)renin Receptor Activation Increases Profibrotic Markers and Fibroblast‐like Phenotype Through MAPK‐dependent ROS Formation In Mouse Renal Collecting Duct Cells.
    Alexis A. Gonzalez, Leonardo Zamora, Cristian Reyes‐Martinez, Nicolas Salinas‐Parra, Nicole Roldan, Catherina A. Cuevas, Stefanny Figueroa, Alex Gonzalez‐Vergara, Minolfa C. Prieto.
    Clinical and Experimental Pharmacology and Physiology. 13 days ago
    Recent studies suggested that activation of the PRR upregulates profibrotic markers through reactive oxygen species (ROS) formation; however, the exact mechanisms have not been investigated in CD cells. We hypothesized that activation of the PRR increases the expression of profibrotic markers through MAPK‐dependent ROS formation in CD cells. Mouse renal CD cell line (M‐1) was treated with recombinant prorenin plus ROS or MAPK inhibitors and PRR‐shRNA to evaluate their effect on the expression of profibrotic markers. PRR immunostaining revealed plasma membrane and intracellular localization. Recombinant prorenin increases ROS formation (6.0 ± 0.5 vs. 3.9 ± 0.1 nM DCF/μg total protein, P<0.05) and expression of profibrotic markers CTGF (149 ± 12%, P<0.05), α‐SMA (160 ± 20%, P<0.05), and PAI‐I (153 ± 13%, P<0.05) at 10‐8 M. Recombinant prorenin induced phospho ERK 1/2 (p44 and p42) at 10‐8 and 10‐6 M after 20 min. Prorenin‐dependent ROS formation and augmentation of profibrotic factors were blunted by ROS scavengers (trolox, p‐coumaric acid, ascorbic acid), the MEK inhibitor PD98059 and PRR transfections with PRR‐shRNA. No effects were observed in the presence of antioxidants alone. Prorenin‐induced upregulation of collagen I and fibronectin was blunted by ROS scavenging or MEK inhibition independently. PRR‐shRNA partially prevented this induction. After 24 h prorenin treatment M‐1 cells undergo to epithelial mesenchymal transition phenotype, however MEK inhibitor PD98059 and PRR knockdown prevented this effect. These results suggest that PRR might have a significant role in tubular damage during conditions of high prorenin‐renin secretion in the CD. This article is protected by copyright. All rights reserved.
    July 11, 2017   doi: 10.1111/1440-1681.12813   open full text
  • Adipose‐derived mesenchymal stem cells therapy for acute kidney injury induced by ischemia‐reperfusion in a rat model.
    Jian‐Bo Zhang, Xiao‐Qiao Wang, Guo‐Lin Lu, Huan‐Sen Huang, Shi‐Yuan Xu.
    Clinical and Experimental Pharmacology and Physiology. July 08, 2017
    Acute kidney injury (AKI) represents a group of complicated syndromes with a high mortality rate. The administration of adipose‐derived mesenchymal stem cells (ADMSCs) has been tested as a possible treatment method for AKI. The long‐term evaluation of AKI induced by ischemia/reperfusion (IR) and the probable renal protection of ADMSCs are limited. In this study we have established a rat AKI model induced by IR and investigated the possible protective effects of ADMSCs. Adult Sprague‐Dawley (SD) rats were divided into three groups (n=6/each group). The MOCK group was as the normal control. Rats in the IR‐AKI and IR‐AKI+ADMSCs groups were subjected to IR injury by clamping both renal pedicles for 40 min. Rats in the MOCK and IR‐AKI groups were injected with PBS via the tail vein as negative treatment controls. Rats in the IR‐AKI+ADMSCs group received ADMSCs therapy (2×106 cells were injected into the rats via the tail vein). We found that ADMSC transplantation restored the pathologic morphology induced by IR‐AKI to normal compared with the MOCK group, suggesting the reparative function of ADMSCs in kidney tissues. Compared with IR‐induced AKI alone, ADMSC treatment significantly decreased the number of apoptotic cells, the level of total urinary protein and serum creatinine, the expression of pro‐inflammatory cytokines (IL‐6, TNF‐α, IL‐1β, IFN‐γ, TNF‐α, IFN‐γ, and TGF‐β), and the inflammation‐associated proteins (HGF and SDF1), but increased the expression of the anti‐inflammatory cytokine, IL‐10, and the anti‐apoptotic regulator, Bcl‐2. Our data have indicated that ADMSC transplantation may protect against IR‐induced AKI by anti‐apoptotic and anti‐inflammatory effects. This article is protected by copyright. All rights reserved.
    July 08, 2017   doi: 10.1111/1440-1681.12811   open full text
  • AQP4‐knockout aggravation of isoprenaline‐induced myocardial injury is mediated by p66Shc and endoplasmic reticulum stress.
    Yusi Cheng, Jie Chao, Dezai Dai, Yin Dai, Dongdong Zhu, Bicheng Liu.
    Clinical and Experimental Pharmacology and Physiology. July 08, 2017
    Aquaporin 4 (AQP4) is a type of water channel protein that maintains the water balance of cardiomyocytes. However, the physiological role of AQP4 in cardiovascular disease is poorly understood. We wanted to explore whether p66Shc and endoplasmic reticulum stress participates in AQP4 knockout (KO)‐mediated cardiac injury. There were two types of mice: AQP4 knockout and wild‐type mice. Each type was randomly divided into three groups: Control group, isoprenaline stimulation group (ISO, 1 mg/kg, s.c., 5 d), and apocynin treatment group (APO, 100 mg/kg, p.o., 3 d). H9c2 rat cardiomyocytes were cultured for RNA interference of AQP4. Results showed increased left ventricular weight index and more severe myocardial inflammation were induced in AQP4 knockout mice relative to wild‐type mice, accompanied by significantly increased levels of the oxidative stress biomarkers MDA and NOX4. In addition, the expressions of p66Shc, ER stress markers PERK, GRP78 and CHOP and proinflammatory factors such as ETA, IL6 and TNFα were upregulated in the myocardium of AQP4 knockout mice or AQP4 siRNA treated cardiomyocytes, whereas CASQ2 was downregulated. ISO stimulation aggravated these abnormalities, which were significantly attenuated by apocynin. This study showed that AQP4 knockout mice were susceptible to cardiac injury induced by ISO. The mechanism was closely connected with p66Shc and proinflammatory factors. Endoplasmic reticulum stress was also involved in the pathological process. This article is protected by copyright. All rights reserved.
    July 08, 2017   doi: 10.1111/1440-1681.12812   open full text
  • Distribution of CYP2C8 and CYP2C9 Amino Acid Substitution Alleles in South Indian Diabetes Patients: A Genotypic and Computational Protein Phenotype Study.
    Durga Koteswara Rao, Dwarakanath K. Murthy, Nazia Sultana Shaik, Babajan Banaganapalli, Kumar Swami Konda, Hanumantha P. Rao, Eswar Ganti, Ahmed A. Zuheir, Ashraf El‐Harouni, Ramu Elango, Imran Ali Khan, Noor Ahmad Shaik.
    Clinical and Experimental Pharmacology and Physiology. July 07, 2017
    The CYP2C8 and CYP2C9 are two major isoforms of the cytochrome P450 enzyme family, which is involved in drug response, detoxification, and disease development. This study describes the differential distribution of amino acid substitution variants of CYP2C8 (*2‐I269F & *3‐R139K) and CYP2C9 (*2‐C144R & *3‐L359A) genes in 234 type 2 diabetes mellitus (T2DM) patients and 218 healthy controls from Andhra Pradesh, South India. Single locus genotype analysis has revealed that homozygous recessive genotypes of 2C8*2‐TT (p=<0.03), 2C9*2‐TT (p=<0.02), and heterozygous 2C9*3‐AC (p=<0.006) are seen to be increasingly present in the case group, indicating a significant level of their association with diabetes in Andhra population. The statistical significance of these recessive genotypes has persisted even under their corresponding allelic forms (p=<0.01). Genotype association results were further examined by computational protein structure and stability analysis to assess the deleteriousness of the amino acid changes. The mutant CYP 2C8 and 2C9 (both *2 and *3) proteins showed structural drifts at both amino acid residue (range 0.43Å‐0.77Å), and polypeptide chain levels (range 0.68Å‐1.81Å) compared to their wild type counterparts. Furthermore, the free energy value differences (range ‐0.915 to ‐1.38 Kcal/mol) between mutant and native protein structures suggests the deleterious and destabilizing potential of amino acid substitution polymorphisms of CYP genes. The present study confirms the variable distribution of CYP2C8 (*2 and *3) and CYP2C9 (*2 and *3) allelic polymorphisms among South Indian diabetic populations and further warrants the serious attention of CYP gene family, as a putative locus for disease risk assessment and therapy. This article is protected by copyright. All rights reserved.
    July 07, 2017   doi: 10.1111/1440-1681.12810   open full text
  • Hypertonic saline inhibits airway smooth muscle contraction by inhibiting Ca2+ sensitization.
    Xiao‐Cao Liu, Qian Wang, Yu‐Shan She, Shu Chen, Xi Luo, Hao Xu, Dun‐An Zang, Wen‐Jing Zhang, Jun‐Ying Qiu, Bei‐Bei Liu, Jinhua Shen, Yong‐Bo Peng, Ping Zhao, Lu Xue, Weiwei Chen, Li‐Qun Ma, Xiangning Fu, Jingyu Chen, Qing‐Hua Liu, Meng‐Fei Yu.
    Clinical and Experimental Pharmacology and Physiology. July 06, 2017
    The effects of hypertonic solution on airway smooth muscle (ASM) contraction and the underlying mechanisms are largely unknown. We found that hypertonic saline (HS) inhibited acetylcholine (ACh)‐induced contraction of ASM from the mouse trachea and human bronchi. In single mouse ASM cells (ASMCs), ACh induced an increase in intracellular Ca2+ that was further enhanced by 5% NaCl, indicating that the HS‐induced inhibition of ASM contraction was not mediated by a decrease in cytosolic Ca2+. The Rho‐associated kinase (ROCK) inhibitor Y‐27632 relaxed ACh‐induced precontraction of mouse tracheal rings. However, such inhibition was not observed after the relaxation induced by 5% NaCl. Moreover, the incubation of mouse tracheal rings with 5% NaCl decreased ACh‐induced phosphorylation of myosin light chain 20 and myosin phosphatase target subunit 1. These data indicate that HS inhibits the contraction of ASM by inhibiting Ca2+ sensitization, not by decreasing intracellular Ca2+. This article is protected by copyright. All rights reserved.
    July 06, 2017   doi: 10.1111/1440-1681.12807   open full text
  • Antitumor bioactive peptides isolated from marine organisms.
    Haibo Xing, Mengting Tong, Nanyu Jiang, Xiaomin Zhang, Hong Hu, Hongming Pan, Da Li.
    Clinical and Experimental Pharmacology and Physiology. July 04, 2017
    Marine organisms are an important source of antitumor active substances. Thus, pharmaceutical research in recent years has focused on exploring new antitumor drugs derived from marine organisms, and, many peptide drugs with strong antitumor activities have been successfully extracted. Based on different mechanisms, this paper reviews the research on several typical antitumor bioactive peptides in marine drugs and the latest progress therein. Additionally, the development prospects for these antitumor bioactive peptide‐based drugs are discussed so as to provide a reference for future research in this field. This article is protected by copyright. All rights reserved.
    July 04, 2017   doi: 10.1111/1440-1681.12808   open full text
  • Down‐regulation of long noncoding RNA MALAT1 by RNA interference inhibits proliferation and induces apoptosis in multiple myeloma.
    Hui Liu, Huihan Wang, Bin Wu, Kun Yao, Aijun Liao, Miao Miao, Yang Li, Wei Yang.
    Clinical and Experimental Pharmacology and Physiology. June 30, 2017
    Multiple myeloma (MM) is a neoplastic plasma‐cell disorder characterized by abnormal proliferation of monoclonal plasma cells in the bone marrow. Metastasis‐associated lung adenocarcinoma transcript 1 (MALAT1), an evolutionarily highly conserved long non‐coding RNA was originally identified in metastatic non‐small cell lung cancer and has been reported to be up‐regulated in many other cancers. However, the function of MALAT1 in MM remains unknown. In the present study, by transfecting MM cells with MALAT1‐specific short hairpin RNA (shRNA) expression plasmids, the role of MALAT1 in the proliferation and apoptosis of MM cells was investigated in vitro, and the tumorigenicity of MALAT1‐silenced cells was evaluated in vivo. MALAT1 was found to be highly expressed in RPMI8226 and U266 cells. Down‐regulation of MALAT1 via RNA interference significantly inhibited the proliferation of MM cells through cell cycle arrest at G1 phase. Moreover, knockdown of MALAT1 induced apoptosis, which was closely associated with the activation of caspase‐3/‐9, down‐regulation of Bcl‐2 and up‐regulation of Bax. In addition, silencing of MALAT1 by intratumoral injection of MALAT1 shRNA attenuated the tumor growth in mice bearing myeloma xenograft and led to massive apoptosis in the xenograft tumor. Therefore, MALAT1 may serve as a promising target in the genetic therapeutic strategy for MM treatment. This article is protected by copyright. All rights reserved.
    June 30, 2017   doi: 10.1111/1440-1681.12804   open full text
  • Lower muscle tissue is associated with higher pulse wave velocity: a systematic review and meta‐analysis of observational study data.
    Alexander J. Rodríguez, Md Nazmul Karim, Velandai Srikanth, Peter Ebeling, David Scott.
    Clinical and Experimental Pharmacology and Physiology. June 28, 2017
    Muscle loss and arterial stiffness share common risk factors and are commonly seen in the elderly. We aimed to synthesise the existing literature on studies that have examined this association. We searched electronic databases for studies reporting correlations or associations between a measure of muscle tissue and a measure of arterial stiffness. Meta‐analysis was conducted using Fisher's Z‐transformed r‐correlation (rZ) values. Pooled weighted rZ and 95% confidence intervals were calculated in an inverse‐variance, random‐effects model. Heterogeneity was assessed by the inconsistency index (I2). Study quality was assessed on a checklist using items from validated quality appraisal guidelines. 1,195 records identified, 21 satisfied our inclusion criteria totalling 8,558 participants with mean age 52±4years (range: 23‐74). Most studies reported an inverse relationship between muscle tissue and arterial stiffness. Eight studies had data eligible for meta‐analysis. Muscle tissue was inversely associated with pulse wave velocity in healthy individuals [rZ = ‐0.15 (95%CI: ‐0.24, ‐0.07); p=0.0006; I2=85%; n=3,577] and in any population [rZ = ‐0.18 (‐0.26, ‐0.10); p<0.0001; I2=81%; n=3,930]. In a leave‐one‐out sensitivity analysis, the results remained unchanged. Lower muscle tissue was associated with arterial stiffness. Studies were limited by cross‐sectional design. Cardiovascular risk monitoring may be strengthened by screening for low muscle mass and maintaining muscle mass may be a primary prevention strategy. This article is protected by copyright. All rights reserved.
    June 28, 2017   doi: 10.1111/1440-1681.12805   open full text
  • Salinomycin Overcomes Acquired Tamoxifen Resistance through AIB1 and Inhibits Cancer Cell Invasion in Endocrine Resistant Breast Cancer.
    Suwisit Manmuan, Nithidol Sakunrangsit, Wannarasmi Ketchart.
    Clinical and Experimental Pharmacology and Physiology. June 28, 2017
    Salinomycin is a monocarboxylic polyether ionophore isolated from Streptomyces albus. It has been widely used as an antibiotic in veterinary medicine in poultry. A recent study demonstrated that salinomycin selectively inhibits human breast cancer stem cells; one possible mechanism of tamoxifen resistance. Our results show that salinomycin is effective in inhibiting MCF‐7/LCC2 and MCF‐7/LCC9 cell lines which are well‐established endocrine resistant cells and has a synergistic effect in combination with tamoxifen using MTT proliferation assay. The inhibitory effect of salinomycin on the reduction of critical ER‐coactivator; amplified breast 1 (AIB1) mRNA and protein expression is overcoming tamoxifen resistance. Moreover, salinomycin significantly inhibits cell invasion in Matrigel invasion assay. The effect was mediated at least in part by the decrease of matrix metalopeptidase 9 (MMP‐9) which is one critical enzyme facilitated in the cell invasion process. In conclusion, salinomycin should be developed as a novel agent used alone or in combination for endocrine‐resistant breast cancer. This article is protected by copyright. All rights reserved.
    June 28, 2017   doi: 10.1111/1440-1681.12806   open full text
  • The use of antioxidant agents for chemotherapy‐induced peripheral neuropathy treatment in animal models.
    Larissa Feitosa Carvalho, Ana Maria Fantini Silva, Adriana Andrade Carvalho.
    Clinical and Experimental Pharmacology and Physiology. June 26, 2017
    Antineoplastic drugs such as Cisplatin, Oxaliplatin, Paclitaxel and Vincristin are widely used in the treatment for several both solid and blood tumours. However, the severity of peripheral neuropathy caused by these agents can affect the patient's quality of life. The major symptoms of Chemotherapy‐Induced Peripheral Neuropathy (CIPN) involve: sensory loss, paresthesia, dysesthaesia, numbness, tingling, temperature sensitivity, allodynia and hyperalgesia, in a “stocking and glove” distribution. Why many different chemotherapeutic agents result in similar neuropathy profiles is unclear. Many drug classes such as antidepressants, anticonvulsants, antispastic agents and others have been used in clinical practice, but there is no scientific evidence to prove their effectiveness. But drugs as the antioxidant have shown a protective effect against free radical damage. Then, in order to find out a successful treatment for CIPN, animal studies (i.e pharmacological and mechanical tests and histopathological immunohistochemical analyses) have been developed to try to determinate the action of the antioxidant agents. This systematic review provides an overview of the major antioxidant agents recently investigated to treat CIPN and the animal models used for this purpose. This article is protected by copyright. All rights reserved.
    June 26, 2017   doi: 10.1111/1440-1681.12803   open full text
  • Ferulic acid alleviates symptoms of preeclampsia in rats by upregulating vascular endothelial growth factor.
    Weiyan Gong, Jipeng Wan, Qing Yuan, Quanzhan Man, Xiaojing Zhang.
    Clinical and Experimental Pharmacology and Physiology. June 22, 2017
    Preeclampsia is a complication affecting pregnant women worldwide, which leads to maternal and fetal morbidity and mortality. In this study, we evaluated the efficacy of ferulic acid (FA) on an Nω‐nitro‐L‐arginine methyl ester hydrochloride (L‐NAME) induced rat model of preeclampsia. L‐NAME was administered to pregnant rats to induce preeclampsia. 48 rats were divided into three experimental groups (n=16 each): control group, preeclampsia group and preeclampsia with FA treatment (preeclampsia+FA). Physiological characteristics such as urine volume, total urine protein and blood pressure were assessed. Expressions levels of urinary nephrin and podocin mRNAs were analyzed by RT‐PCR. Levels of renal vascular endothelial growth factor (VEGF), renal soluble fms‐like tyrosine kinase‐1 (sFlt‐1) and serum placenta growth factor (PlGF) were also examined. Urine volume, total urine protein and blood pressure were markedly increased in preeclampsia group rats compared to control (p<0.05), which were then significantly reduced in preeclampsia+FA group (p<0.05). Expressions of urinary nephrin and podocin mRNAs, levels of VEGF, sFlt‐1 and PlGF were also reversed in preeclampsia+FA group compared to preeclampsia rats (p<0.05). We hereby report for the first time, FA alleviates preeclampsia symptoms in a rat preeclampsia model, supporting its potential value in treating preeclampsia. This article is protected by copyright. All rights reserved.
    June 22, 2017   doi: 10.1111/1440-1681.12801   open full text
  • A co‐drug conjugate of naringenin and lipoic acid mediates neuroprotection in a rat model of oxidative stress.
    Tarek M Saleh, Monique C Saleh, Barry J Connell, Yang‐Heon Song.
    Clinical and Experimental Pharmacology and Physiology. June 21, 2017
    Using our in vitro and in vivo models of oxidative stress, the current study was designed to determine the neuroprotective potential of naringenin, alone or in combination with lipoic acid. In our mixed neuronal culture exposed to hypoxia and subsequent reoxygenation, naringenin was shown to provide significant neuroprotection against cell death at a concentration of 2.5 μM. Lipoic acid (LA) did not produce neuroprotection at any concentration tested (0.25 to 100 μM). In contrast, when naringenin was covalently combined with LA, producing a novel compound named “VANL‐100”, significant neuroprotection was observed at a concentration as low as 2 x 10‐2 μM (100‐fold more potent). An ELISA for antioxidant capacity demonstrated that naringenin and VANL‐100 likely resulted in neuroprotection by increasing the free radical scavenging capacity of the neuronal cells. Pretreatment of rats with the above compounds prior to middle cerebral artery occlusion (MCAO) followed by reperfusion, showed similar results. Naringenin significantly reduced infarct volume at a dose of 10 mg/kg while VANL‐100 produced significant neuroprotection at a dose as low as 1x10‐4 mg/kg (10,000‐fold more potent). This VANL‐100‐induced neuroprotection persisted even when administered 1 and 3 hours into the reperfusion time course. Taken together, these results suggest that our novel compound, VANL‐100 is neuroprotective, likely via a mechanism that involves increasing the antioxidant capacity of neuronal cells. Our results also show that VANL‐100 is 100 to 10,000‐fold more potent than the parent compounds, which adds to the growing evidence in support of combination therapy targeting oxidative stress in neurodegenerative diseases. This article is protected by copyright. All rights reserved.
    June 21, 2017   doi: 10.1111/1440-1681.12799   open full text
  • Targeting protein for xenopus kinesin‐like protein 2 knockdown enhances radiation sensitivity of human lung squamous carcinoma cell.
    Jie Yang, Feng Gao, Xinjian Xu, Yuxiang Wang, Shuchai Zhu.
    Clinical and Experimental Pharmacology and Physiology. June 21, 2017
    Targeting protein for Xenopus kinesin‐like protein 2 (TPX2) has been demonstrated to be associated with the tumorigenesis of many cancers. In the present study, we investigated the role and preliminary mechanism of TPX2 in the resistance of lung squamous carcinoma to radiation therapy. The results showed that SK‐MES‐1R and NCI‐H226R cells were more resistant to X‐ray irradiation than the parental cells (SK‐MES‐1 and NCI‐H226). Moreover, TPX2 was upregulated in the radioresistant cells compared with the parental cells. TPX2 knockdown significantly decreased TPX2 expression in SK‐MES‐1 cells, while TPX2 overexpression increased TPX2 expression in NCI‐H226 cells compared with the corresponding control cells. TPX2 knockdown enhanced the radiosensitivity of SK‐MES‐1 and promoted cell apoptosis following exposure to irradiation, whereas TPX2 overexpression decreased the radiosensitivity of NCI‐H226 and inhibited cell apoptosis. In in vivo studies, the combination of TPX2 knockdown and irradiation significantly inhibited tumor growth, decreased tumor weight, downregulated TPX2 expression in tumor tissue and induced cell apoptosis in nude mice, while TPX2 overexpression exerted an opposite effect. Our results indicated that TPX2 was correlated with cell radioresistance and it might be served as a therapeutic target to enhance cell radiosensitivity in the radiation therapy of lung squamous carcinoma. This article is protected by copyright. All rights reserved.
    June 21, 2017   doi: 10.1111/1440-1681.12800   open full text
  • Immunosuppressive efficacy of tetrandrine combined with methylprednisolone against mitogen‐activated peripheral blood mononuclear cells of hemodialysis patients.
    Wencheng Xu, Kehan Meng, Junichi Kusano, Hiroto Matsuda, Yoshikazu Hara, Yoshiaki Fujii, Shinya Suzuki, Eiki Ando, Xiaoqin Wang, Yuanchao Tu, Sachiko Tanaka, Kentaro Sugiyama, Haruki Yamada, Toshihiko Hirano.
    Clinical and Experimental Pharmacology and Physiology. June 14, 2017
    Immunosuppressive therapy for prevention of acute rejection episode occasionally causes serious adverse effects, and thus it is important to develop new therapeutic approach for renal transplant recipients. This study evaluated the immunosuppressive pharmacodynamics of tetrandrine (TET) and/or methylprednisolone (MP) in hemodialysis patients in vitro by using the peripheral blood mononuclear cells (PBMCs) isolated from whole blood of hemodialysis patients. The median (range) of MP IC50 values against the proliferation of patients PBMCs was 7.04 (2.30~500.00) ng/mL. In contrast, the median (range) of MP IC50 values against the proliferation of healthy PBMCs was 4.44 (3.19~5.08) ng/mL. The median (range) of TET IC50 values against the proliferation of patients PBMCs was 1.61 (1.04~4.79) μM. Lower concentrations of TET (0.3‐300 nM) were able to decrease the IC50 values of MP and thus potentiate the MP immunosuppressive effect on patient PBMCs. The median (range) of MP IC50 values in combination with 0.3, 3, 30, and 300 nM TET were 0.92 (0.49~8.39), 2.10 (0.45~20.00), 0.35 (0.092~1.05), and 0.14 (0.05~6.78) ng/mL, respectively. TET potentiates the MP immunosuppressive pharmacodynamics and thus, it was possible to use the combination of MP and TET to attenuate MP side effects. There were significant correlations between the IC50 values of TET and stimulation indices (P=0.04, r=0.58), the IC50 values of TET and the hemodialysis periods (P=0.04, r=0.57), or the IC50 values of MP combined with 0.3 nM TET and C‐reactive protein concentrations (P=0.04, r=0.64), respectively. This article is protected by copyright. All rights reserved.
    June 14, 2017   doi: 10.1111/1440-1681.12797   open full text
  • Effect of iloprost on biomarkers in patients with congenital heart disease–pulmonary arterial hypertension.
    Xiao‐ye Li, Yu Zheng, Yuliang Long, Xiaochun Zhang, Lei Zhang, Dan Tian, Daxin Zhou, Qian‐zhou Lv.
    Clinical and Experimental Pharmacology and Physiology. June 13, 2017
    Some biomarkers play important roles in the endothelial dysfunction of patients with pulmonary arterial hypertension (PAH), including nitric oxide (NO), endothelin‐1 (ET‐1), asymmetric dimethylarginine (ADMA), galectin‐3 (Gal‐3), B‐type natriuretic peptide (BNP), and uric acid (UA). However, studies on these biomarkers in pulmonary artery blood in congenital heart disease–PAH (CHD–PAH) and the effect of iloprost on the regulation of biomarkers are lacking. This study investigated potential CHD–PAH biomarkers and their association with the severity of disease. The effect of iloprost on the regulation of these biomarkers was also studied. A total of 31 patients with CHD–PAH were enrolled. 7 with positive effects of iloprost (the average reduction in mPAP 11.13 ± 1.73 mm Hg) and 19 with negative effects of iloprost [the average reduction in mPAP 4.21 ± 4.87 mm Hg, iloprost positive group (IPG) vs iloprost negative group (ING), P < 0.01]—and 5 age‐matched controls were studied. The pulmonary artery blood sample was collected before and after inhaling iloprost, and the plasma concentrations of Gal‐3, ADMA, ET‐1, and NO were measured. A significant positive linear relationship was observed between mPAP and plasma ET‐1, BNP, ADMA, and UA levels in all patients with CHD–PAH. ET‐1, ADMA, BNP, and UA levels had a significant linear relationship with mean pulmonary arterial pressure, which could be used to predict the severity of CHD–PAH. ET‐1 might be a potential biomarker to pre‐evaluate the effect of iloprost on CHD–PAH. Iloprost could affect the expression of Gal‐3 and, therefore, the process of fibrosis could be influenced by iloprost. This article is protected by copyright. All rights reserved.
    June 13, 2017   doi: 10.1111/1440-1681.12796   open full text
  • Roles of partitioning‐defective protein 6 (Par6) and its complexes in the proliferation, migration and invasion of cancer cells.
    Lingling Ruan, Yanting Shen, Ziwen Lu, Dongsheng Shang, Zhicong Zhao, Yongjin Lu, Yanfang Wu, Yafei Zhang, Zhigang Tu, Hanqing Liu.
    Clinical and Experimental Pharmacology and Physiology. June 07, 2017
    A pivotal regulator of cell polarity and homeostasis, partitioning‐defective protein 6 (Par6) forms multicomponent complexes that not only regulate cell polarity and stabilize cell morphology, but have also been demonstrated to participate in the proliferation, migration and invasion of cancer cells. The transforming growth factor (TGF)‐β and extracellular signal‐regulated kinase (Erk) 1/2 pathways are the most thoroughly studied pathways involving Par6 in many cancers. Aurothiomalate has been used to disrupt the interaction between Par6 and atypical protein kinase C within the multicomponent complexes, and has been shown to effectively block transformed growth and metastasis in vitro and/or in vivo in a variety of cancers, including pancreatic, prostate and lung cancers, as well as alveolar rhabdomyosarcoma. It is likely that with further revelations regarding the critical roles of Par6 in cancer initiation, progression and metastasis, targeted therapies against Par6 will be discovered and prove effective preclinically, and hopefully clinically, in cancer treatment. This article is protected by copyright. All rights reserved.
    June 07, 2017   doi: 10.1111/1440-1681.12794   open full text
  • Probiotic Bifidobacterium bifidum G9‐1 attenuates 5‐fluorouracil‐induced intestinal mucositis in mice via suppression of dysbiosis‐related secondary inflammatory responses.
    Shinichi Kato, Nahla Hamouda, Yoshitaro Kano, Yousuke Oikawa, Yoshiki Tanaka, Kenjiro Matsumoto, Kikuko Amagase, Masaki Shimakawa.
    Clinical and Experimental Pharmacology and Physiology. June 07, 2017
    Bifidobacterium, a major component of the intestinal microbiota, has been clinically used for the treatment of diarrhoea and constipation. 5‐Fluorouracil (5‐FU), widely used for cancer chemotherapy, is known to frequently induce intestinal mucositis accompanied by severe diarrhoea. The present study examined the effect of Bifidobacterium bifidum G9‐1 (BBG9‐1) on 5‐FU‐induced intestinal mucositis in mice. Intestinal mucositis was induced by repeated administration of 5‐FU for 6 days. BBG9‐1 was administered orally once daily for 9 days, beginning 3 days before the onset of 5‐FU treatment. Repeated administration of 5‐FU caused severe intestinal mucositis, characterised by shortening of villi and destruction of crypts, accompanied by increases in intestinal myeloperoxidase activity and inflammatory cytokine expression, body weight loss, and diarrhoea on day 6. Daily administration of BBG9‐1 significantly reduced the severity of intestinal mucositis and inflammatory responses and tended to attenuate clinical symptoms. In contrast, BBG9‐1 failed to prevent apoptosis induction on day 1 after the first 5‐FU administration. The structure of the intestinal microbiota, as analysed by weighted UniFrac distance, was largely altered by 5‐FU treatment, but this change was mitigated by daily administration of BBG9‐1. Moreover, 5‐FU treatment decreased the abundance of Firmicutes and increased the abundance of Bacteroidetes, but these responses were also significantly inhibited by daily administration of BBG9‐1. These results suggest that BBG9‐1 has an ameliorative effect against 5‐FU‐induced intestinal mucositis through the attenuation of inflammatory responses via improve dysbiosis. BBG9‐1 could be useful for the prevention of intestinal mucositis during cancer chemotherapy. This article is protected by copyright. All rights reserved.
    June 07, 2017   doi: 10.1111/1440-1681.12792   open full text
  • Dexmedetomidine protects mice against myocardium ischemic/reperfusion injury by activating an AMPK/PI3K/Akt/eNOS pathway.
    Yanjun Sun, Chuan Jiang, Jun Jiang, Lisheng Qiu.
    Clinical and Experimental Pharmacology and Physiology. May 29, 2017
    Acute myocardial ischemia/reperfusion (MIR) injury leads to severe arrhythmias and has a high rate of lethality. In the present study, we aim to determine the effect of dexmedetomidine (Dex) on heart injury parameters following MIR surgery. We examined the effects of Dex on heart function parameters and infarct size following MIR surgery. Proinflammatory cytokines, oxidative products and anti‐oxidative enzymes in the myocardium were measured to evaluate the anti‐inflammatory and anti‐oxidative effects of Dex. The role of the adenosine 5’‐monophosphate (AMP)‐activated protein kinase (AMPK)/phosphatidylino‐sitol 3‐kinase (PI3k)/Akt/endothelial nitric oxide synthase (eNOS) pathway was investigated using their inhibitors. The alteration of hemodynamic parameters, histopathological results, and infarct size caused by MIR was attenuated by Dex. The interleukine‐1 beta (IL‐1β), IL‐6, tumor necrosis factor‐a (TNF‐α) and myeloperoxidase (MPO) were all significantly decreased. Anti‐oxidative enzymes superoxide dismutase (SOD), catalase and glutathione peroxidase (GPx) were restored by Dex. Oxidative products8‐OHdG, MDA and protein carbonyl were all decreased by Dex (p<0.05). Dex activated AMPK expression, eNOS and Akt phosphorylation. The influence of Dex on cardiac function was reversed by the inhibitors of the eNOS, AMPK and PI3K/Akt pathways. These results indicate that Dex protected the cardiac functional, histological changes, inflammation and oxidative stress induced by MIR. Our results present a novel signaling mechanism that Dex protects MIR injury by activating an AMPK/PI3K/Akt/eNOS pathway. This article is protected by copyright. All rights reserved.
    May 29, 2017   doi: 10.1111/1440-1681.12791   open full text
  • Spinal SET7/9 may contribute to the maintenance of cancer‐induced bone pain in mice.
    Li‐Hua Hang, Zhen‐Kai Xu, Shi‐You Wei, Wei‐Wei Shu, Hong Luo, Jian Chen.
    Clinical and Experimental Pharmacology and Physiology. May 29, 2017
    Cancer‐induced bone pain (CIBP) profoundly influences patients’ quality of life. Exploring the mechanisms by which CIBP occurs is essential for developing efficacious therapies. Various studies have shown that proinflammatory factors were involved in CIBP. SET domain containing lysine methyltransferase 7/9 (SET7/9) may modulate the expression of NF‐κB‐dependent proinflammatory genes in vitro. However, whether SET7/9 may participate in the maintenance of CIBP remains unknown. In this study, NCTC 2472 cells were inoculated into the intramedullary space of the femur to establish a mouse model of CIBP. Upregulation of spinal SET7/9 expression was related to pain behaviours in tumour‐inoculated mice. Intrathecal cyproheptadine (10 or 20 nmol) attenuated response to painful stimuli in a dose‐dependent manner. Moreover, there was a concomitant decrease in spinal SET7/9 and RANTES expression. The antinociceptive effects of cyproheptadine were abolished by pre‐intrathecal administration of SET 7/9 (0.2 μg) for 30 min before intrathecal cyproheptadine (20 nmol) administration. These results indicated that spinal SET7/9 may contribute to the maintenance of CIBP in mice. Hence, targeting of spinal SET7/9 might be a useful alternative therapy for the treatment of CIBP. This article is protected by copyright. All rights reserved.
    May 29, 2017   doi: 10.1111/1440-1681.12789   open full text
  • Antiarrhythmic properties of ivabradine in an experimental model of Short‐QT‐ Syndrome.
    Gerrit Frommeyer, Jan Weller, Christian Ellermann, Sven Kaese, Simon Kochhäuser, Philipp S. Lange, Dirk G. Dechering, Lars Eckardt.
    Clinical and Experimental Pharmacology and Physiology. May 29, 2017
    The If channel inhibitor ivabradine is recommended for treatment of chronic heart failure. However, ivabradine also inhibits human ether‐a‐go‐go (hERG) mediated potassium currents. The aim of the present study was to assess the electrophysiologic effects of ivabradine in an experimental model of short‐QT‐syndrome. 12 rabbit hearts were isolated and Langendorff‐perfused. After obtaining baseline data, pinacidil, an IK‐ATP channel opener, was infused (1μM). Eight endo‐ and epicardial monophasic action potentials and a 12‐lead ECG showed a significant abbreviation of QT interval (‐32ms,p<0.05) and shortening of action potential duration at 90% of repolarization (APD90; ‐22ms, p<0.05). The shortening of ventricular repolarization was accompanied by a reduction of effective refractory period (ERP;‐20ms,p<0.05). Thereafter, hearts were additionally treated with ivabradine (5μM) leading to an increase of QT‐interval (+31ms,p<0.05), APD90 (+15ms,p<0.05) as well as of ERP (+38ms,p<0.05) and post‐repolarization refractoriness (PRR,+33ms,p<0.05) as compared with sole pinacidil infusion. Under baseline conditions, ventricular fibrillation (VF) was inducible by a standardized pacing protocol including programmed stimulation and burst stimulation in 3 of 12 hearts (6 episodes). After application of 1μM pinacidil, 6 of 12 hearts were inducible (22 episodes). Additional infusion of 5μM ivabradine led to a significant suppression of VF. Only 2 episodes could be induced in 1 of 12 hearts. In the present study ivabradine reversed the electrophysiologic effects of pharmacologically simulated short‐QT syndrome. Ivabradine demonstrated antiarrhythmic properties based on an increase of both ERP and PRR. This article is protected by copyright. All rights reserved.
    May 29, 2017   doi: 10.1111/1440-1681.12790   open full text
  • Moderate‐to‐high normal levels of thyrotropin is a risk factor for urinary incontinence and an unsuitable quality of life in women over 65 years.
    Estela Cuevas‐Romero, Angélica Sánchez‐Cardiel, Angélica M. Zamora‐Gallegos, Rosalía Cruz‐Lumbreras, Dora L. Quintanilla, Francisco Castelán, Margarita Martínez‐Gómez.
    Clinical and Experimental Pharmacology and Physiology. May 28, 2017
    The present study was aimed to investigate the relationship between normal serum concentrations of thyrotropin (TSH) and urinary incontinence (IU), urinary infections, and quality of life in old women. Euthyroid post‐menopausal women without sarcopenia, estrogen replacement, emotional illness, and‐or cancer were enrolled as participants. Anthropometric indicators, serum glucose and estradiol, and thyroid profile were measured. Sociodemographic, clinical, physical activity, and quality of life (SF‐36) surveys were applied. One‐hour pad test and International Consultation on Incontinence Questionnaire Short Form (ICIQ‐SF) were used to determine UI. Urinalysis was also done. In agreement with results from the pad test (cut‐off point ≥ 1.4 g), the ICIQ‐SF reveled ~50% of incontinent women. A high percentage of women had moderate‐high bacteriuria and urinary infections. Logistic regression analysis showed that age is a risk factor for both UI and urinary infection. Nor diabetes, number of pregnancies or childbirths, urinary infections, and bacteriuria influenced in the presence of UI. To allocate women into four groups according to their age (<65 or ≥65 years old) and TSH concentrations (0.3‐1.9 or 2‐10 μUI/mL), we found that moderate‐to‐high normal levels of TSH is a risk factor for UI and a worst quality of life in the oldest women. Our results highlight the profit of measuring TSH concentrations in post‐menopausal women. This article is protected by copyright. All rights reserved.
    May 28, 2017   doi: 10.1111/1440-1681.12788   open full text
  • Neuroprotective effects of a novel poly (ADP‐ribose) polymerase‐1 inhibitor, JPI‐289, in hypoxic rat cortical neurons.
    Youngchul Kim, Young S. Kim, Min‐Young Noh, Hanchang Lee, Boyoung Joe, Hyun Y. Kim, Jeongmin Kim, Seung H. Kim, Jiseon Park.
    Clinical and Experimental Pharmacology and Physiology. May 19, 2017
    Excessive activation of poly (ADP‐ribose) polymerase‐1 (PARP‐1) is known to develop neuronal apoptosis, necrosis and inflammation after ischaemic brain injury. Therefore, PARP‐1 inhibition after ischaemic stroke has been attempted in successful animal studies. The purpose of present work was to develop a novel water soluble PARP‐1 inhibitor (JPI‐289) and explore its neuroprotective effect on ischaemic injury in an in vitro model. The half‐life of JPI‐289 after intravenous or oral administration in rats was relatively long (1.4‐1.5 hours) with 65.6% bioavailability. The inhibitor strongly inhibited PARP‐1 activity (IC50=18.5 nmol/L) and cellular PAR formation (IC50=10.7 nmol/L) in the nanomolar range. In rat cortical neuronal cells, JPI‐289 did not affect cell viability up to 1 mmol/L as assayed by Trypan blue staining (TBS) and lactate dehydrogenase (LDH) assay. Treatment of JPI‐289 for 2 hours after 2 hours of oxygen glucose deprived (OGD) rat cortical neuron attenuated PARP activity and restored ATP and NAD+ levels. Apoptosis‐associated molecules such as apoptosis inducing factor (AIF), cytochrome C and cleaved caspase‐3 were reduced after JPI‐289 treatment in the OGD model. The present findings suggest that the novel PARP‐1 inhibitor, JPI‐289, is a potential neuroprotective agent which could be useful as a treatment for acute ischaemic stroke.
    May 19, 2017   doi: 10.1111/1440-1681.12757   open full text
  • Air pollution and short‐term clinical outcomes of patients with acute myocardial infarction.
    Min Woo Lee, Byoung Geol Choi, Suhng Wook Kim, Seung‐Woon Rha, Min Suk Shim, Dae Jin Kim, Hong Seog Seo, Dong Joo Oh, Myung ho Jeong,.
    Clinical and Experimental Pharmacology and Physiology. May 19, 2017
    Ambient air pollution is well‐known to be a serious risk factor for cardiovascular diseases, stroke, and death. However, the association between air pollutants (AP) exposure and short‐term clinical outcomes in acute myocardial infarction (AMI) patients (pts) has not been elucidated well. In the present study, 37 880 AMI pts were enrolled from October 2005 to December 2013 in a nationwide large‐scale, prospective, multicentre Korea AMI registry (KAMIR registry; We obtained data on AP (e.g., NO2, SO2, CO, O3 and PM10) from the Korean National Institute of Environmental Research (NIER; Clinical endpoints included death, recurrent myocardial infarction (Re‐MI), any revascularization and composite of all‐cause death and Re‐MI. Exposure to AP is defined as the average exposure to AP within 24 hours before AMI admission. We observed that a 0.01 part per million (ppm) increase in NO2 concentration, 0.001 ppm increase in SO2 concentration, and 0.1 ppm increase in CO concentration each increased the risk of total death by 9.7% (95% CI, 6.2%‐13.4%), 1.9% (95% CI, 0.3%‐3.6%), and 2.1% (95% CI, 0.5%‐3.9%), respectively. Exceptionally, O3 decreased the risk of total death by 0.6% (95% CI −0.2% to −1.0%) per 0.01 ppm increase. PM10 was not related to any cardiovascular events. AP were each stratified into five quintiles according to ranges of AP levels. After adjusting analysis for risk variables, only high quintiles (Q4, Q5) of NO2 were positively associated with total death, cardiac death and MI, while SO2, CO, O3 and PM10 were shown to be not related to any cardiovascular events at all levels. In AMI patients, each AP and its concentration has shown a different effect to short‐term mortality and cardiovascular events.
    May 19, 2017   doi: 10.1111/1440-1681.12755   open full text
  • INPP4B overexpression suppresses migration, invasion and angiogenesis of human prostate cancer cells.
    Haiwen Chen, Hongliang Li, Qi Chen.
    Clinical and Experimental Pharmacology and Physiology. May 19, 2017
    Inositol polyphosphate 4‐phosphatase B (INPP4B) has been identified as a tumour suppressor in different human cancers. However, the role of INPP4B in the angiogenesis of human prostate cancer cells remains unclear. In this study, we first compared the expression of INPP4B between prostate cancer tissues and tumour‐adjacent normal prostate tissues using immunohistochemistry. Then, we explored the role of INPP4B in prostate cancer progression via transfection of a Flag‐INPP4B plasmid into PC3 and DU145 cells in vitro and in vivo. Our results showed that reduced INPP4B staining was significantly correlated with the tumour‐node‐metastasis stage. Moreover, transfection with Flag‐INPP4B plasmid suppressed the migration and invasion of prostate cancer cells through inactivating the PI3K/Akt signalling pathway, at the same time decreased vascular endothelial growth factor secretion and suppressed human umbilical vein endothelial cells proliferation and tube formation. Futhermore, it was also found that INPP4B could inhibit tumour growth and angiogenesis in vivo. Altogether, our results supported that INPP4B acted as a tumour suppressor in human prostate cancer, and provided insights into development of a targeted therapy for this disease.
    May 19, 2017   doi: 10.1111/1440-1681.12745   open full text
  • OligoG CF‐5/20 normalizes cystic fibrosis mucus by chelating calcium.
    Anna Ermund, Christian V Recktenwald, Gudmund Skjåk‐Bræk, Lauren N Meiss, Edvar Onsøyen, Philip D Rye, Arne Dessen, Astrid Hilde Myrset, Gunnar C Hansson.
    Clinical and Experimental Pharmacology and Physiology. May 19, 2017
    The goal of this study was to determine whether the guluronate (G) rich alginate OligoG CF‐5/20 (OligoG) could detach cystic fibrosis (CF) mucus by calcium chelation, which is also required for normal mucin unfolding. Since bicarbonate secretion is impaired in CF, leading to insufficient mucin unfolding and thereby attached mucus, and since bicarbonate has the ability to bind calcium, we hypothesized that the calcium chelating property of OligoG would lead to detachment of CF mucus. Indeed, OligoG could compete with the N‐terminus of the MUC2 mucin for calcium binding as shown by microscale thermophoresis. Further, effects on mucus thickness and attachment induced by OligoG and other alginate fractions of different length and composition were evaluated in explants of CF mouse ileum mounted in horizontal Ussing‐type chambers. OligoG at 1.5% caused effective detachment of CF mucus and the most potent alginate fraction tested, the poly‐G fraction of about 12 residues, had similar potency compared to OligoG whereas mannuronate‐rich (M) polymers had minimal effect. In conclusion, OligoG binds calcium with appropriate affinity without any overt harmful effect on the tissue and can be exploited for treating mucus stagnation.
    May 19, 2017   doi: 10.1111/1440-1681.12744   open full text
  • Development of resistance to serotonin‐induced itch in bile duct ligated mice.
    Sattar Ostadhadi, Nazgol‐Sadat Haddadi, Arash Foroutan, Ehsan Azimi, Sarina Elmariah, Ahmad‐Reza Dehpour.
    Clinical and Experimental Pharmacology and Physiology. May 19, 2017
    Cholestatic itch can be severe and significantly impair the quality of life of patients. The serotonin system is implicated in cholestatic itch; however, the pruritogenic properties of serotonin have not been evaluated in cholestatic mice. Here, we investigated the serotonin‐induced itch in cholestatic mice which was induced by bile duct ligation (BDL). Serotonin, sertraline or saline were administered intradermally to the rostral back area in BDL and sham operated (SHAM) mice, and the scratching behaviour was videotaped for 1 hour. Bile duct ligated mice had significantly increased scratching responses to saline injection on the seventh day after surgery. Additionally, serotonin or sertraline significantly induced scratching behaviour in BDL mice compared to saline at day 7 after surgery, while it did not induce itch at day 5. The scratching behaviour induced by serotonin or sertraline was significantly less in BDL mice compared to SHAM mice. Likewise, the locomotor activity of BDL or SHAM mice was not significantly different from unoperated (UNOP) mice on the fifth and seventh day, suggesting that the scratching behaviour was not affected by motor dysfunctions. Our data suggest that despite the potentiation of evoked itch, a resistance to serotonin‐induced itch is developed in cholestatic mice.
    May 19, 2017   doi: 10.1111/1440-1681.12752   open full text
  • Arrhythmic effects of Epac‐mediated ryanodine receptor activation in Langendorff‐perfused murine hearts are associated with reduced conduction velocity.
    Mengye Li, Sandeep S Hothi, Samantha C Salvage, Kamalan Jeevaratnam, Andrew A Grace, Christopher L‐H Huang.
    Clinical and Experimental Pharmacology and Physiology. May 19, 2017
    Recent papers have attributed arrhythmic substrate in murine RyR2‐P2328S hearts to reduced action potential (AP) conduction velocities (CV), reflecting acute functional inhibition and/or reduced expression of sodium channels. We explored for acute effects of direct exchange protein directly activated by cAMP (Epac)‐mediated ryanodine receptor‐2 (RyR2) activation on arrhythmic substrate and CV. Monophasic action potential (MAP) recordings demonstrated that initial steady (8 Hz) extrinsic pacing elicited ventricular tachycardia (VT) in 0 of 18 Langendorff‐perfused wild‐type mouse ventricles before pharmacological intervention. The Epac activator 8‐CPT (8‐(4‐chlorophenylthio)‐2′‐O‐methyladenosine‐3′,5′‐cyclic monophosphate) (VT in 1 of 7 hearts), and the RyR2 blocker dantrolene, either alone (0 of 11) or with 8‐CPT (0 of 9) did not then increase VT incidence (P>.05). Both progressively increased pacing rates and programmed extrasystolic (S2) stimuli similarly produced no VT in untreated hearts (n=20 and n=9 respectively). 8‐CPT challenge then increased VT incidences (5 of 7 and 4 of 8 hearts respectively; P<.05). However, dantrolene, whether alone (0 of 10 and 1 of 13) or combined with 8‐CPT (0 of 10 and 0 of 13) did not increase VT incidence relative to those observed in untreated hearts (P>.05). 8‐CPT but not dantrolene, whether alone or combined with 8‐CPT, correspondingly increased AP latencies (1.14±0.04 (n=7), 1.04±0.03 (n=10), 1.09±0.05 (n=8) relative to respective control values). In contrast, AP durations, conditions for 2:1 conduction block and ventricular effective refractory periods remained unchanged throughout. We thus demonstrate for the first time that acute RyR2 activation reversibly induces VT in specific association with reduced CV.
    May 19, 2017   doi: 10.1111/1440-1681.12751   open full text
  • Potential excitatory role of nitric oxide on 2‐deoxy‐d‐glucose‐induced gastric motility in rats.
    Ayse M. Sevgili, Dicle Z. Balkanci, Aysen Erdem.
    Clinical and Experimental Pharmacology and Physiology. May 19, 2017
    Previous studies have shown that 2‐deoxy‐d‐glucose (2‐DG) increases gastric motility via the vagus nerve, but the underlying mechanism remains elusive. Since nitric oxide (NO) is involved in gastric motility, a possible interplay between 2‐DG and NO can be suggested. In the present study, Wistar rats (250‐350 g) of both sexes were intravenously injected with 2‐DG (200 mg/kg), and the effects of the intravenous injection of the nitric oxide synthase (NOS) inhibitors; nitro‐l‐arginine methyl ester (l‐NAME, 10 mg/kg) and Nω‐nitro‐l‐arginine (l‐NNA, 10 mg/kg) were investigated. Animals were anaesthetized and cannulated for intravenous drug injections while the left vagal nerve was electrically stimulated (0.1‐10 Hz, 0.5 ms duration, 12 V, for 60 seconds), and intragastric pressure and gastric motility changes were monitored using a latex gastric balloon. 2‐DG increased the mean intragastric pressure (baseline, 5.0±0.4 cmH2O; after 2‐DG, 14.4±1.5 cmH2O; P=.0156) and significantly increased the gastric motility index, while NOS inhibitors significantly attenuated both parameters. However, pretreatment with NOS inhibitors significantly augmented the gastric responses to peripheral electrical vagal stimulation. These results suggest that NO plays an excitatory role in gastric responsiveness to 2‐DG and that this function may be effected in the central nervous system.
    May 19, 2017   doi: 10.1111/1440-1681.12749   open full text
  • Changes in PPARδ expression in a rat model of stress‐induced depression.
    Keng‐Fan Liu, Yingxiao Li, Kai Chun Cheng, Chao Chin Hsu, Juei‐Tang Cheng, Wen‐Huang Peng.
    Clinical and Experimental Pharmacology and Physiology. May 19, 2017
    Depression is a common mental disorder that has been linked to a decrease in the expression of serotonin and/or the serotonin transporter in the brain. Antidepressants that target the monoaminergic system are widely used in the clinical setting. Peroxisome proliferator‐activated receptor δ (PPAR δ) overexpression or activation is thought to improve depression‐like behaviours in rodents. The present study was designed to characterize the changes in PPARδ expression in the hippocampus in rats with stress‐induced depression. We used an unpredictable chronic mild stress (CMS) model in rats to study the role of PPARδ in the hippocampus. Behaviour was evaluated via a forced swim test (FST), a tail suspension test (TST), and a sucrose preference test (SPT). Then, the changes in PPARδ expression and other signals were determined using Western blots. We found that PPARδ expression in the hippocampus was markedly reduced in rats with depression. Moreover, the expression of the serotonin transporter was also significantly decreased. Treatment with a PPARδ agonist enhanced the expression of PPARδ and the serotonin transporter in the hippocampus of rats with stress‐induced depression. Additionally, treatment with a PPARδ agonist increased the expression of the serotonin transporter in cultured hippocampal (H19‐7) cells, and this action was ablated in the absence of PPARδ, which was attenuated with shRNA. Taken together, we found that PPARδ plays an important role in the regulation of serotonin transporter expression and that chronic stress may lower PPARδ expression in the brain via apoptosis and may attenuate serotonin transporter expression, thus inducing depression in rats.
    May 19, 2017   doi: 10.1111/1440-1681.12746   open full text
  • Gastric cancer management: Kinases as a target therapy.
    Batoul Farran, Susanne Müller, Raquel C Montenegro.
    Clinical and Experimental Pharmacology and Physiology. May 19, 2017
    The molecular diagnostics revolution has reshaped the practice of oncology by facilitating the identification of genetic, epigenetic and proteomic modifications correlated with cancer, thus delineating ‘oncomaps’ for various cancer types. These advances have enhanced our understanding of gastric cancer, one of the most fatal diseases worldwide, and culminated in the approval of novel molecular therapies such as trastuzumab. Gastric tumours display recurrent aberrations in key kinase oncogenes such as Her2, epidermal growth factor receptor (EGFR), PI3K, mTOR or c‐Met, suggesting that these receptors are amenable to inhibition using specific drug agents. In this review, we examine the mutational landscape of gastric cancer, the use of kinase inhibitors as targeted therapies in gastric tumours and the clinical trials underway for novel inhibitors, highlighting successes, failures and future directions.
    May 19, 2017   doi: 10.1111/1440-1681.12743   open full text
  • Limb remote ischaemic postconditioning‐induced elevation of fibulin‐5 confers neuroprotection to rats with cerebral ischaemia/reperfusion injury: Activation of the AKT pathway.
    Wei Zhang, Ye Wang, Guorong Bi.
    Clinical and Experimental Pharmacology and Physiology. May 19, 2017
    Limb remote ischaemic postconditioning (RIPostC) is an effective and well‐acknowledged treatment for brain ischaemia injury. The present study aimed to evaluate the role of fibulin‐5 in the neuroprotection of RIPostC against cerebral ischaemia/reperfusion (I/R) injury in rats. The middle cerebral artery occlusion (MCAO) model was established in rats and then RIPostC was carried out by three cycles of 10 minutes occlusion/10 minutes release of the bilateral femoral artery at the beginning of the reperfusion. To downregulate the fibulin‐5 level, fibulin‐5 siRNA was injected into the lateral ventricle 24 hours before MCAO. According to our present study, RIPostC attenuated cerebral I/R injury by decreasing infarct volume, improving neurobehavioral score and suppressing blood brain barrier (BBB) leakage. Moreover, the mRNA and protein levels of fibulin‐5 were upregulated by RIPostC at 24 hours and 72 hours after reperfusion. Downregulation of fibulin‐5 attenuated the neuroprotection of RIPostC. Finally, the result showed that fibulin‐5 was upregulated by RIPostC via activation of the PI3K/AKT pathway. Taken together, these results provide evidence that upregulation of fibulin‐5 is involved in the beneficial effect of RIPostC against cerebral I/R injury.
    May 19, 2017   doi: 10.1111/1440-1681.12742   open full text
  • Metformin attenuates renal fibrosis in both AMPKα2‐dependent and independent manners.
    Yenan Feng, Shuaixing Wang, Youyi Zhang, Han Xiao.
    Clinical and Experimental Pharmacology and Physiology. May 19, 2017
    Metformin is a well‐known AMP‐activated protein kinase (AMPK) activator, and it has been shown to inhibit organ fibrosis. Whether AMPKα2 mediates metformin protection against renal fibrosis remains unknown. Here, we aimed to investigate the role of the AMPKα2 isoform in mediating the inhibitory effect of metformin on renal fibrosis. Unilateral ureteral obstruction (UUO) was used to induce renal fibrosis in wild‐type (WT) and AMPKα2 knockout (AMPKα2−/−) mice. Metformin treatment was initiated 3 days before UUO and was continued until 7 days after UUO. In WT mice, metformin significantly inhibited UUO‐induced renal fibrosis. In AMPKα2−/− mice, metformin also tended to inhibit UUO‐induced renal fibrosis. Specifically, metformin significantly reduced UUO‐induced transforming growth factor β1 (TGFβ1) mRNA and protein expression in WT mice but not in AMPKα2−/− mice. In contrast, metformin reduced UUO‐induced TGFβ1 downstream Smad3 phosphorylation in both WT and AMPKα2−/− mice, suggesting that this regulation occurs in an AMPKα2‐independent manner. In conclusion, the underlying mechanisms for the protective effects of metformin against renal fibrosis include AMPKα2‐dependent targeting of TGFβ1 production and AMPKα2‐independent targeting of TGFβ1 downstream signalling. In this regard, metformin has an advantage over other AMPK activators for the treatment of renal fibrosis.
    May 19, 2017   doi: 10.1111/1440-1681.12748   open full text
  • Association of genetic polymorphisms of claudin‐1 with small vessel vascular dementia.
    Vivek Srinivasan, Nady Braidy, Ying Hua Xu, Peter Xie, Kiran Kancherla, Sashiruben Chandramohan, Eunice Kar Wing Chan, Daniel KY Chan.
    Clinical and Experimental Pharmacology and Physiology. May 19, 2017
    The most recent hypothesis of the development of small vessel vascular dementia (VaD) emphasises the role of blood‐brain barrier (BBB) dysfunction. It is hypothesised that certain genetic polymorphisms of the BBB tight junction claudin‐1 protein, in combination with adverse environmental risk factors, increase the risk of BBB dysfunction and small vessel VaD. In this case‐control study, 97 control participants, with a mean Mini Mental State Exam (MMSE) score of 29.1, and 38 VaD participants were recruited and completed a questionnaire on their medical history and lifestyle factors. Blood was also collected and two single nucleotide polymorphisms (SNPs), rs17501010 and rs893051 of claudin‐1 genotyping, were analysed by real‐time polymerase chain reaction (PCR) assay. A significantly higher frequency of all rs893051 SNP genotypes (GC and CC) was found in the VaD population (OR=4.8, P=0.006 and OR=6, P<0.001 respectively). Patients with TT genotype of rs17501010 were also more likely to have VaD (OR=3.25, P=0.022). Stratification analysis revealed that having combined haplotype GC+CC of rs893051 and lipid disorders was associated with higher risk of VaD (OR=9.9, P<0.001). For patients with type 2 diabetes the odds ratio of VaD increased significantly in GC+CC genotypes of rs893051 (OR=12.57, P<0.0001) and GT+TT of rs17501010 (OR=5.33, P=0.01).
    May 19, 2017   doi: 10.1111/1440-1681.12747   open full text
  • An investigation of the anti‐inflammatory effects and a potential biomarker of PI3Kδ inhibition in COPD T cells.
    Abid Khan, Thomas Southworth, Sally Worsley, Srividya Sriskantharajah, Augustin Amour, Edith M. Hessel, Dave Singh.
    Clinical and Experimental Pharmacology and Physiology. May 15, 2017
    Lymphocyte numbers are increased in the lungs of chronic obstructive pulmonary disease (COPD) patients. Phosphatidylinositol‐3‐kinase delta (PI3Kδ) is involved in lymphocyte activation. We investigated the effect of PI3Kδ inhibition on cytokine release from COPD lymphocytes. We also evaluated phosphorylated ribosomal S6 protein (rS6) as a potential biomarker of PI3Kδ activation. Peripheral blood mononuclear cells (PBMCs) and bronchoalveolar lavage (BAL) cells isolated from healthy never smokers (HNS), smokers (S) and COPD patients were stimulated to induce a T cell receptor response. The effects of a PI3Kδ specific inhibitor (GSK045) on cytokine release and rS6 phosphorylation were measured by Luminex and flow cytometry respectively. The effects of GSK045 on cytokine production from PHA stimulated chopped lung samples were investigated. GSK045 reduced cytokine release from PBMCs, BAL cells and chopped lung. Inhibition was greatest in the chopped lung model, with approximately 80% inhibition of IFNγ, IL‐2, IL‐17 and IL‐10. PI3Kδ inhibition suppressed rS6 phosphorylation in unstimulated airway T‐lymphocytes by up to 60%. Inhibition of PI3Kδ suppressed T cell cytokine production in COPD patients. rS6 phosphorylation shows potential as a biomarker to assess PI3Kδ activity. This article is protected by copyright. All rights reserved.
    May 15, 2017   doi: 10.1111/1440-1681.12784   open full text
  • A novel synthetic chemical entity (UPEI‐800) is neuroprotective in vitro and in an in vivo rat model of oxidative stress.
    Tarek M. Saleh, Monique C. Saleh, Barry J. Connell, Inan Kucukkaya, Alaa S. Abd‐El‐Aziz.
    Clinical and Experimental Pharmacology and Physiology. May 15, 2017
    In this study, we tested a novel synthetic pyrazole‐containing compound, 5‐amino‐1‐phenyl‐1H‐pyrazole‐4‐carbonitrile (APPC), as an antioxidant in both in vitro and in vivo models of oxidative stress. In addition, the utility of covalently combining APPC with another well‐established antioxidant, lipoic acid (LA), was also tested in both models. The in vitro results demonstrated that pretreatment with APPC in a mixed neuronal‐glial culture exposed to oxygen‐glucose deprivation (OGD) followed by reoxygenation‐refeeding, resulted in significant neuroprotection at concentrations between 2.5 to 25 μM. In contrast, LA was not neuroprotective following OGD alone or following reoxygenation‐refeeding. However, the synthetic covalent combination of APPC with LA, named “UPEI‐800″, resulted in significant neuroprotection at concentrations between 0.027 and 2.7 μM (100‐fold more potent than APPC alone), an effect shown to be correlated with increased cellular antioxidant capacity. Further, in an in vivo model of ischemia‐reperfusion injury following transient occlusion of the middle cerebral artery (tMCAO), both APPC (0.1 and 1.0 mg/kg) and UPEI‐800 (1 x 10‐3 mg/kg) provided significant neuroprotection. Consistent with the in vitro findings, the in vivo results following tMCAO also demonstrated a 100‐fold increase in the potency of the covalently linked compound UPEI‐800 compared to APPC alone. This article is protected by copyright. All rights reserved.
    May 15, 2017   doi: 10.1111/1440-1681.12785   open full text
  • Inhibitory effect of the urothelium/lamina propria on female porcine urethral contractility & effect of age.
    Oladayo Seun Folasire, Russ Chess‐Williams, Donna Jayne Sellers.
    Clinical and Experimental Pharmacology and Physiology. May 06, 2017
    The urethral uroepithelium has been implicated in urethral sensation and maintenance of continence. However, relatively little is known about the function of the urethral urothelium compared with that of the bladder. The aim of the study was to examine the role of the urothelium/lamina propria on contractility of the porcine urethra, along with the influence of nitric oxide, prostaglandins and ageing. Porcine urethral tissues, intact and denuded of urothelium/lamina propria, were mounted in tissue baths and contractions to noradrenaline, phenylephrine and carbachol obtained. Contractions in the presence of Nώ–nitro‐L‐arginine (100μM) and indomethacin (10μM) were examined, along with contractions of tissues from young (6 months) and older (3 years) animals. The urothelium/lamina propria of the urethra significantly inhibited contractions to carbachol, noradrenaline and phenylephrine. This inhibitory effect was not significantly different for the three agonists (58.7±10.3%, 60.4±12.6% and 39.4±12.2% inhibition, n=4‐7), and was also observed when denuded tissues were co‐incubated with a second tissue with intact urothelium/lamina propria (40.6±7.5% inhibition, n=6). Inhibition of nitric oxide and prostaglandin production did not attenuate the inhibitory effect of the urothelium/lamina propria on noradrenaline contractions. In addition, ageing did not alter the inhibitory effect for either phenylephrine contractions (33.9±2.2% vs 41.0±9.7%, young vs older urethral tissues) or noradrenaline contractions (32.9±11.1% vs 53.7±11.0%). In conclusion the urothelium/lamina propria of the urethra has an inhibitory effect on receptor‐mediated urethral contraction. This inhibition is due to the release of a diffusible factor, and the effect is not mediated by nitric oxide or prostaglandins, or affected by age. This article is protected by copyright. All rights reserved.
    May 06, 2017   doi: 10.1111/1440-1681.12779   open full text
  • MicroRNA regulation of extracellular matrix components in the process of atherosclerotic plaque destabilization.
    Michal Kowara, Agnieszka Cudnoch‐Jedrzejewska, Grzegorz Opolski, Pawel Wlodarski.
    Clinical and Experimental Pharmacology and Physiology. April 25, 2017
    The process of atherosclerotic plaque destabilization, leading to myocardial infarction, is still not fully understood. The pathway – composed of structural and regulatory proteins of the extracellular matrix (ECM) such as collagen, elastin, small leucine‐rich proteoglycans, metalloproteinases, cathepsins and serine proteases – is one potential way of atherosclerotic plaque destabilization. The expression of these proteins is controlled by different microRNA molecules. The goal of this paper is to summarize the current investigations and knowledge about ECM in the process of atherosclerotic plaque destabilization, giving special attention to epigenetic expression regulation by microRNA. This article is protected by copyright. All rights reserved.
    April 25, 2017   doi: 10.1111/1440-1681.12772   open full text
  • Mechanism of the vasorelaxant effect induced by trans‐4‐methyl‐β‐nitrostyrene, a synthetic nitroderivative, in rat thoracic aorta.
    Taylena Maria Teófilo, Loeste Arruda‐Barbosa, Jussara Mathyelle Rodrigues‐Silva, Joyce Karen Lima Vale, Rosivaldo dos Santos Borges, Gloria Pinto Duarte, Pedro Jorge Caldas Magalhães, Saad Lahlou.
    Clinical and Experimental Pharmacology and Physiology. April 22, 2017
    Mechanisms underlying the vasorelaxant effects of trans‐4‐methyl‐β‐nitrostyrene (T4MeN) were studied in rat aortic rings. In endothelium‐intact preparations, T4MeN fully and similarly relaxed contractions induced by phenylephrine (PHE) (IC50 = 61.41 [35.40‐87.42] μM) and KCl (IC50 = 83.50 [56.63‐110.50] μM). The vasorelaxant effect of T4MeN was unchanged by endothelium removal, pretreatment with L‐NAME, indomethacin, tetraethylammonium, ODQ or MDL‐12,330A. Under Ca2+‐free conditions, T4MeN significantly reduced with a similar potency: (i) phasic contractions induced by PHE, but not by caffeine; (ii) contractions due to CaCl2 in aortic preparations stimulated with PHE (in the presence of verapamil) or high KCl; (iii) contractions evoked by the restoration of external Ca2+ levels after depletion of intracellular Ca2+ stores in the presence of thapsigargin. In contrast, T4MeN was more potent at inhibiting contractions evoked by the tyrosine phosphatase inhibitor, sodium orthovanadate, than those induced by the activator of PKC, phorbol‐12,13‐dibutyrate. These results suggest that T4MeN induces an endothelium‐ independent vasorelaxation that appears to occur intracellularly through the inhibition of contractions that are independent of Ca2+ influx from the extracellular milieu but involve phosphorylation of tyrosine residues. This article is protected by copyright. All rights reserved.
    April 22, 2017   doi: 10.1111/1440-1681.12771   open full text
  • Comparative proteomic analysis of urine and laser microdissected glomeruli in IgA nephropathy.
    Xiaoyuan Ning, Zhong Yin, Zuoxiang Li, Jiayun Xu, Linna Wang, Wanjun Shen, Yang Lu, Guangyan Cai, Xueguang Zhang, Xiangmei Chen.
    Clinical and Experimental Pharmacology and Physiology. April 21, 2017
    The purpose of the present study was to identify the differential proteins that synchronously change in urine and glomeruli and could be used to monitor glomerular lesions of IgA nephropathy (IgAN). The proteomes of urine and glomeruli from four IgAN patients who were graded III/IV according to the grading system of Lee et al. were compared to the urine proteomes of four healthy volunteers and the glomeruli proteomes of adjacent normal tissue from four patients with renal tumors, respectively. Western blot, enzyme‐linked immunosorbent assay and immunofluorescence assay were applied to verify the results of the proteomic analysis. In the proteomic analysis of urine from IgAN patients and healthy volunteers, 714 proteins were identified, with 246 proteins identified as differential proteins. In the proteomic analysis of glomeruli from renal biopsy tissue of IgAN patients and from adjacent normal tissue of patients with renal tumors, 161 proteins were identified altogether, and 20 proteins of these were recognized as differential proteins. After comparatively analyzing the differential proteins identified in the urine and glomeruli, five synchronously changed differential proteins were found: complement C9, Ig kappa chain C region and three cytoskeleton proteins. In summary, our findings indicate that certain immunological indices in urine appear to be associated with glomerular lesions of IgAN.
    April 21, 2017   doi: 10.1111/1440-1681.12733   open full text
  • The intrauterine renin–angiotensin system: Sex‐specific effects on the prevalence of spontaneous preterm birth.
    Kirsty G Pringle, Tamas Zakar, Eugenie R Lumbers.
    Clinical and Experimental Pharmacology and Physiology. April 21, 2017
    Preterm birth (PTB) is the single largest cause of death in infants and young children. The rate of PTB is significantly higher in male infants, particularly those that are born very preterm. Here we present evidence to suggest that the decidual renin–angiotensin system may play a role in inhibiting inflammation and maintaining the integrity of the fetal membranes during pregnancy, and that sex‐specific alterations in the intrauterine RAS could contribute to the increased risk of PTB in male babies. Women carrying female fetuses have high levels of expression of decidual prorenin at term. Decidua from ‘female’ pregnancies also have greater expression of the anti‐inflammatory angiotensin (Ang)‐(1‐7) pathway, than decidua from ‘male’ pregnancies, and have lower levels of the pro‐inflammatory Ang II pathway. We propose that in ‘female’ pregnancies, the very high levels of decidual prorenin drive the anti‐inflammatory Ang‐(1‐7) pathway, thus reducing the likelihood of PTB. In addition, the high levels of prorenin produced by the decidua in ‘female’ pregnancies are able to diffuse into the amnion and bind to the PRR. We postulate that PRR/prorenin interactions, possibly through both angiotensin dependent and independent pathways, stimulate the production of ECM proteins, inhibit ECM degradation and prevent apoptosis, thus strengthening the amnion. Thus control of the inflammatory signature and the integrity of the fetal membranes prior to parturition may partly depend on the sexually determined activity of the decidual and amniotic renin–angiotensin system pathways.
    April 21, 2017   doi: 10.1111/1440-1681.12734   open full text
  • Reduced inflammatory responses of follicular helper T cell promote the development of regulatory B cells after Roux‐en‐Y gastric bypass.
    Junfang Zhan, Liyu Huang, Haiyong Ma, Huan Chen, Yuan Yang, Sheng Tan, Wendy Song, Weiguo Zhao, Xiaojiang Dai.
    Clinical and Experimental Pharmacology and Physiology. April 21, 2017
    Bariatric surgery is currently the most effective strategy in treating severe obesity and its comorbidities, such as type 2 diabetes (T2D). However, the mechanism through which bariatric surgery mediates its benefits is not completely understood. Since obesity and T2D represent yet another inflammatory disease, and follicular helper T (Tfh) cells play important roles in inflammatory disorders, we investigated whether the Tfh activity was altered after Roux‐en‐Y gastric bypass (RYGB), one of the most common bariatric surgery procedures. We found that the Tfh cells after RYGB were not significantly changed in number, but presented altered cytokine secretion profile, including lower interferon (IFN)‐γ, interleukin (IL)‐2, IL‐4, and IL‐17 secretion. Tfh cells after RYGB also downregulated inducible co‐stimulator and programmed death‐1. Interestingly, after Tfh cell‐naive B cell coculture, Tfh cells after RYGB secreted more IL‐10 than autologous Tfh cells before RYGB. The frequencies of IL‐10‐expressing and transforming growth factor (TGF)‐β‐expressing regulatory B cells after Tfh cell‐naive B cell coculture were directly correlated with the frequency of IL‐10‐expressing Tfh cells. Depletion of IL‐10 in the coculture, however, resulted in fewer regulatory B cells. Finally, patients with greater increase in IL‐10‐expressing Tfh cells presented further reductions in body mass index, glycaemia, and body fat percentage. Together, these data demonstrated that the Tfh cells after RYGB presented lower inflammatory status and secreted higher IL‐10, through which these Tfh cells promoted the development of regulatory B cells. Higher IL‐10‐expressing Tfh cell level also predicted better patient response to RYGB.
    April 21, 2017   doi: 10.1111/1440-1681.12740   open full text
  • Impact of polymorphisms in angiogenesis‐related genes on clinical outcomes of radiotherapy in patients with nasopharyngeal carcinoma.
    Wan‐Le Ma, Rong Liu, Li‐Hua Huang, Chan Zou, Jie Huang, Jing Wang, Shao‐Jun Chen, Xiang‐Guang Meng, Jing‐Ke Yang, Han Li, Guo‐Ping Yang, Cheng‐Xian Guo.
    Clinical and Experimental Pharmacology and Physiology. April 21, 2017
    The purpose of this paper is to assess the relationship between gene polymorphism in angiogenesis‐related genes and radiation responses in nasopharyngeal carcinoma (NPC) patients. The genotypes of 180 NPC patients were analyzed by Sequenom MassARRAY. The response evaluation criteria in solid tumours were used for assessing efficacies, and the criteria of the Radiation Therapy Oncology Group or European Organization for Research & Treatment of Cancer were utilized for evaluating acute toxic reactions in response to radiation. Statistical methods included chi‐square test, uni‐ and multivariate logistic regression analyses. Genotypic carriers of rs1800541 GT were at an elevated risk of developing grade 3+ oral mucositis, and a genetic variant of rs5333 was a predictor for a lower occurring risk of grade 2+ radiation‐induced xerostomia. EDN1 rs1800541, rs2071942 and rs5370 variants were associated with a significantly higher risk of severe myelosuppression. SNPs in such angiogenesis‐related genes as EDN1 rs1800541, rs2071942 & rs5370 and EDNRA rs5333 may serve as useful biomarkers for predicting the outcomes of NPC patients.
    April 21, 2017   doi: 10.1111/1440-1681.12738   open full text
  • Population pharmacokinetics analysis of intravenous busulfan in Chinese patients undergoing hematopoietic stem cell transplantation.
    Xuemei Wu, Helin Xie, Weiwei Lin, Ting Yang, Nainong Li, Shanshan Lin, Xiaohong Yuan, Jinhua Ren, Xiaofan Li, Xian Huang.
    Clinical and Experimental Pharmacology and Physiology. April 21, 2017
    There are several reports describing population pharmacokinetic (popPK) models of busulfan (BU). However, limited information is available in Chinese hematopoietic stem cell transplantation (HSCT) patients. The present study aimed to establish a popPK model of intravenous BU in Chinese HSCT patients for individualized drug therapy. The popPK model of BU was developed from a total of 284 concentration–time points from 53 patients. The effects of demographic and biochemical covariates were investigated by nonlinear mixed effect model (NONMEM) software. Plots, visual predictive check (VPC), bootstrap and normalized prediction distribution error (NPDE) were performed to determine the stability and the reliability of the final model. A one‐compartment model with first‐order elimination process was confirmed as the final structural model for BU. For a typical patient whose body surface area (BSA) is 1.7 m2, the population typical values of CL and Vd were 11.86 L/h, and 48.2 L, respectively. The result suggested BSA showed significant influence on CL and Vd (P<.001). Plots revealed the final model was performing a goodness fit. The steady rate verified by bootstrap was 100%, relative deviation was less than 4.00%, estimated value of final model was in the 95% confidence interval (CI). The VPC results showed the observed values were almost all positioned within the 5th and 95th CIs. The mean and variance of the NPDE were 0.0363 (Wilcoxon signed‐rank test, 0.298) and 0.877 (Fisher variance test, 0.134; SW test of normality, 0.108), respectively. The global adjusted P value was 0.305, which indicated that the prediction of the BU popPK model was adequate. A physician‐friendly Microsoft Excel‐base tool was implemented using the final popPK model for designing individualized dosing regimens.
    April 21, 2017   doi: 10.1111/1440-1681.12735   open full text
  • Palmitoleic acid reduces the inflammation in LPS‐stimulated macrophages by inhibition of NFκB, independently of PPARs.
    Camila O Souza, Alexandre AS Teixeira, Luana A Biondo, Loreana S Silveira, Philip C Calder, José C Rosa Neto.
    Clinical and Experimental Pharmacology and Physiology. April 21, 2017
    Palmitoleic acid (PM, 16:1n‐7) has anti‐inflammatory properties that could be linked to higher expression of PPARα, an inhibitor of NFκB. Macrophages play a major role in the pathogenesis of chronic inflammation, however, the effects of PM on macrophages are underexplored. Thus, we aimed to investigate the effects of PM in activated macrophages as well the role of PPARα. Primary macrophages were isolated from C57BL/6 wild type (WT) and PPARα knockout (KO) mice, cultured under standard conditions and exposed to lipopolysaccharides LPS (2.5 μg/ml) and PM 600 μmol/L conjugated with albumin for 24 hours. The stimulation with LPS increased the production of interleukin (IL)‐6 and IL‐1β while PM decreased the production of IL‐6 in WT macrophages. In KO macrophages, LPS increased the production of tumour necrosis factor (TNF)‐α and IL‐6 and PM decreased the production of TNFα. The expression of inflammatory markers such NFκB and IL1β were increased by LPS and decreased by PM in both WT and KO macrophages. PM reduced the expression of MyD88 and caspase‐1 in KO macrophages, and the expression of TLR4 and HIF‐1α in both WT and KO macrophages, although LPS had no effect. CD86, an inflammatory macrophage marker, was reduced by PM independently of genotype. PM increased PPARγ and reduced PPARβ gene expression in macrophages of both genotypes, and increased ACOX‐1 expression in KO macrophages. In conclusion, PM promotes anti‐inflammatory effects in macrophages exposed to LPS through inhibition of inflammasome pathway, which was independent of PPARα, PPARϒ and AMPK, thus the molecular mechanisms of anti‐inflammatory response caused by PM is still unclear.
    April 21, 2017   doi: 10.1111/1440-1681.12736   open full text
  • Effect of parthanatos on ropivacaine‐induced damage in SH‐SY5Y cells.
    Ting Zheng, Chun‐ying Zheng, Xiao‐chun Zheng, Ruo‐guang Zhao, Yan‐qing Chen.
    Clinical and Experimental Pharmacology and Physiology. April 21, 2017
    Ropivacaine is one of the most common but toxic local anaesthetics, and the mechanisms underlying its neurotoxicity are still largely unknown. This study was conducted to prepare a ropivacaine‐induced neuronal injury model and research the effects of ropivacaine on PARP‐1 activation and nicotinamide adenine dinucleotide (NAD)+ depletion. The cell death and apoptosis of ropivacaine‐induced SH‐SY5Y cells were detected with flow cytometry. The lactate dehydrogenase cycling reaction measured the NAD+ level, and western blots were used to analyze the expression levels of PARP‐1 and apoptosis‐inducing factor (AIF) after ropivacaine treatments with different concentrations and durations. A PARP‐1 inhibitor (PJ‐34) was used to confirm the relationship between PARP‐1 activation and NAD+ depletion. Hoechst 33258 nuclear staining and a mitochondrial membrane potential (Δψm) assay were used to detect the role of exogenous NAD+ in ropivacaine‐induced neuronal injury. Ropivacaine‐induced SH‐SY5Y cell death and apoptosis, PARP‐1 activation, and AIF increase as well as intracellular NAD+ depletion occurred in a time‐ and concentration‐dependent manner (P<.05). PARP‐1 activation led to NAD+ depletion (P<.05). Exogenous NAD+ impaired ropivacaine‐induced nuclear injury (P<.05). Ropivacaine treatment induced PARP‐1 activation and NAD+ depletion (P<.05). Parthanatos (PARP‐1‐dependent cell death) was definitely involved in ropivacaine‐induced neuronal injury, and exogenous NAD+ may be a novel therapeutic method for parthanatos‐dependent neuronal injury.
    April 21, 2017   doi: 10.1111/1440-1681.12730   open full text
  • Inhibition of ALDH2 protects PC12 cells against formaldehyde‐induced cytotoxicity: involving the protection of hydrogen sulphide.
    Ying Chen, Cheng‐Fang Zhou, Fan Xiao, Hong‐Lin Huang, Ping Zhang, Hong‐Feng Gu, Xiao‐Qing Tang.
    Clinical and Experimental Pharmacology and Physiology. April 21, 2017
    Formaldehyde (FA), a common environmental contaminant, has toxic effects on the central nervous system (CNS). We have previously found that hydrogen sulphide (H2S), the third endogenous gaseous mediator, protects neuron against the toxicity of FA. However, the underlying mechanism is poor. Aldehyde‐dehydrogenase‐2 (ALDH2) plays a major role in detoxification of reactive aldehyde in a range of organs and cell types. Therefore, we speculated that H2S antagonizes FA‐induced neurotoxicity by modulating ALDH2. In the present study, we found that the exposure of PC12 cells to FA causes increase in ALDH2 expression and activity. Daidzin, an inhibitor of ALDH2, significantly antagonizes FA‐exerted cytotoxicity and oxidative stress including the accumulation of intracellular reactive oxygen species (ROS), 4‐hydroxy‐2‐trans‐nonenal (4‐HNE), and malondialdehyde (MDA), in PC12 cells. We also showed that daidzin markedly attenuated FA‐induced apoptosis in PC12 cells. Furthermore, we found that H2S reverses FA‐elicited upregulation of ALDH2 in PC12 cells. Our results demonstrated the involvement of downregulation of ALDH2 in the protection of H2S against FA neurotoxicity.
    April 21, 2017   doi: 10.1111/1440-1681.12741   open full text
  • Antihyperglycaemic action of diosmin, a citrus flavonoid, is induced through endogenous β‐endorphin in type I‐like diabetic rats.
    Chia‐Chen Hsu, Mang Hung Lin, Juei Tang Cheng, Ming Chang Wu.
    Clinical and Experimental Pharmacology and Physiology. April 21, 2017
    Diosmin is one of the flavonoids contained in citrus and has been demonstrated to improve glucose metabolism in diabetic disorders. However, the mechanism(s) of diosmin in glucose regulation remain obscure. Therefore, we investigated the potential mechanism(s) for the antihyperglycaemic action of diosmin in streptozotocin‐induced diabetic rats (STZ‐diabetic rats). Diosmin lowered hyperglycaemia in a dose‐dependent manner in STZ‐diabetic rats. This action was inhibited by naloxone at a dose sufficient to block opioid receptors. Additionally, we determined the changes in plasma β‐endorphin‐like immunoreactivity (BER) using enzyme‐linked immunosorbent assay (ELISA). Diosmin also increased BER dose‐dependently in the same manner. Repeated treatment of STZ‐diabetic rats with diosmin for 1 week resulted in an increase in the expression of the glucose transporter subtype 4 (GLUT 4) in the soleus muscle and a reduction in the expression of phosphoenolpyruvate carboxykinase (PEPCK) in the liver. These effects were also inhibited by naloxone at a dose sufficient to block opioid receptors. Bilateral adrenalectomy in STZ‐diabetic rats eliminated the actions of diosmin, including both the reduction in hyperglycemia and the elevation of plasma BER. In conclusion, our results suggest that diosmin may act on the adrenal glands to enhance the secretion of β‐endorphin, which can stimulate the opioid receptors to attenuate hepatic gluconeogenesis and increase glucose uptake in soleus muscle, resulting in reduced hyperglycemia in STZ‐diabetic rats.
    April 21, 2017   doi: 10.1111/1440-1681.12739   open full text
  • Silencing of astrocyte elevated gene‐1 inhibits proliferation and migration and induces apoptosis in melanoma.
    Yue Zhang, Ge Peng, Ying Wang, Lixia Cui, Wenqing Wu, Luan Wang, Chengyu Liu, Xiuping Han.
    Clinical and Experimental Pharmacology and Physiology. April 21, 2017
    Melanoma is an aggressive skin malignancy with a high mortality. Astrocyte elevated gene‐1 (AEG‐1), a downstream target of ras and c‐myc, has been implicated in the development of multiple tumors, yet its role in melanoma remains unclear. In the present study, the role of AEG‐1 in melanoma was explored through AEG‐1 silencing. Our results showed that silencing of AEG‐1 inhibited proliferation of melanoma cells and induced cell cycle arrest, accompanied by reduced levels of cyclinA, cyclinB, cyclinD1, cyclinE and cyclin dependent kinase 2. AEG‐1 silencing also induced apoptosis in melanoma cells and altered the levels of cleaved caspase‐3, Bax and Bcl‐2. Moreover, silencing of AEG‐1 suppressed migration and invasion of melanoma cells, and reduced the expressions and activities of MMP‐2 and MMP‐9. In addition, the activation of Wnt/β‐catenin signaling pathway in melanoma cells was inhibited by AEG‐1 silencing. Furthermore, in vivo experiments revealed that AEG‐1 silencing inhibited the growth of melanoma xenografts in nude mice. In summary, our study demonstrates an oncogenic role of AEG‐1 in melanoma and suggests that AEG‐1 may serve as a potential therapeutic target in the treatment of melanoma. This article is protected by copyright. All rights reserved.
    April 21, 2017   doi: 10.1111/1440-1681.12767   open full text
  • Involvement of ERK1/2 in Cx43 depression induced by macrophage migration inhibitory factor in atrial myocytes.
    Xin Li, Fang Rao, Chun‐yu Deng, Wei Wei, Fang‐zhou Liu, Hui Yang, Zhao‐yu Wang, Su‐juan Kuang, Xiao‐yan Chen, Yu‐mei Xue, Shu‐lin Wu.
    Clinical and Experimental Pharmacology and Physiology. April 21, 2017
    Connexin 43 (Cx43) plays an important role in the pathogenesis of atrial fibrillation (AF). The present study sought to investigate the effect of macrophage migration inhibitory factor (MIF), a pleiotropic cytokine, on Cx43 expression and activity and determine the intracellular signaling pathways. Cx43 protein and mRNA levels were assayed using immunofluorescence, real‐time PCR, and western blot. We found that increased MIF and extracellular regulated protein kinases (ERK) expression was accompanied by a significant reduction in Cx43 protein expression in atrial tissues from patients with AF compared with those with sinus rhythm. In cultured atrium‐derived myocytes (HL‐1 cells), mouse recombinant‐MIF (rMIF, 20 or 40 nM, 24 h) down‐regulated gene and protein expression of Cx43 in a concentration‐dependent manner. U0126, a specific inhibitor of mitogen‐activated protein kinase kinase (MAPKK) could reverse the decrease in expression of Cx43 protein induced by rMIF. Further studies revealed that rMIF (40 nM, 15, 30, and 45 min) was able to stimulate phospho‐Erk1/2 (Thr202/Tyr204) production in a time‐dependent manner. These results suggest that MIF is involved in the pathogenesis of AF, probably by down‐regulating the protein and gene expression of Cx43 via ERK1/2 kinase activation. Our findings represent a potential pathogenic mechanism in AF. This article is protected by copyright. All rights reserved.
    April 21, 2017   doi: 10.1111/1440-1681.12766   open full text
  • Anti‐inflammatory and retinal protective effects of capsaicin on ischemia‐induced injuries through the release of endogenous somatostatin.
    Jun Wang, Wenke Tian, Shuaiwei Wang, Wenqiang Wei, Dongdong Wu, Honggang Wang, Li Wang, Ruisheng Yang, Ailing Ji, Yanzhang Li.
    Clinical and Experimental Pharmacology and Physiology. April 21, 2017
    The mechanisms regarding the retinal protective and anti‐inflammatory effects of capsaicin (CAP) remain unclear. Somatostatin is contained in CAP‐sensitive sensory neurons, including nerve terminals, from which it can be released by capsaicin. The present study provides a novel neurohumoral regulatory mechanism for CAP‐induced‐endogenous somatostatin in a retinal ischemic‐reperfusion (I/R) mouse model. CAP (0.5 mg kg−1) was injected subcutaneously 5 min after I/R. A selective somatostatin‐depleting agent, cysteamine, was applied subcutaneously 4 hours before the experiment to examine the effects of endogenous somatostatin. Ischemia and oxidative stress‐induced inflammatory factors (CXCL10, CXCR3 and NF‐κB p65) were also examined in the present study. The morphometric evaluation showed that the retinal thickness was increased 24 hours after I/R injury and attenuated 7 days after I/R injury. The number of ganglion cells was reduced 7 days after I/R injury. The application of CAP significantly prevented retinal I/R damage. Cysteamine pretreatment reversed the effects of CAP. Inhibition of CXCL10/CXCR3 and NF‐κB (especially in astrocytes and microglia/macrophage) was involved in capsaicin‐induced retinal protection through endogenous somatostatin. CAP has anti‐inflammatory and neuroprotective effects in ischemia‐induced retinal injuries through endogenous somatostatin. Novel therapeutic remedies for inflammation or neuronal injuries were developed based on the systemic humoral effects related to CAP. This article is protected by copyright. All rights reserved.
    April 21, 2017   doi: 10.1111/1440-1681.12769   open full text
  • Clarification of serotonin‐induced effects in peripheral artery disease observed through the femoral artery response in models of diabetes and vascular occlusion: the role of calcium ions.
    Marko Stojanović, Milica Prostran, Radmila Janković, Miroslav Radenković.
    Clinical and Experimental Pharmacology and Physiology. April 21, 2017
    Recent findings have demonstrated that serotonin is an important participant in the development and progression of peripheral artery diseases. Taking this into consideration, the goals of this study were to investigate the effects of serotonin on isolated Wistar rat femoral arteries in both healthy and diabetic animals, with and without artery occlusion, with a particular focus on determining the role of calcium in this process. Contraction experiments with serotonin on intact and denuded femoral artery rings, in the presence or absence of nifedipine and ouabain (both separately, or in combination), as well as Ca2+‐free Krebs–Ringer bicarbonate solution were performed. The serotonin‐induced results were concentration dependent, but only in healthy animals. The endothelium‐dependent contraction of the femoral artery was assessed. In healthy animals, the endothelium‐reliant part of contraction was dependent on the extracellular calcium, while the smooth muscle‐related part was instead dependent on the intracellular calcium. In diabetic animals, both nifedipine and ouabain influenced serotonin‐induced vascular effects by blocking intracellular calcium pathways. However, this was diminished after the simultaneous administration of both blockers. This article is protected by copyright. All rights reserved.
    April 21, 2017   doi: 10.1111/1440-1681.12770   open full text
  • NDUFA4L2 protects against ischemia/reperfusion‐induced cardiomyocyte apoptosis and mitochondrial dysfunction by inhibiting complex I.
    Jianhua Li, Caiyan Bai, Junxia Guo, Wanqian Liang, Jingning Long.
    Clinical and Experimental Pharmacology and Physiology. April 21, 2017
    Myocardial ischemia/reperfusion (I/R) injury may cause the apoptosis of cardiomyocytes as well as mitochondrial dysfunction. The aims of the present study were to investigate whether NADH dehydrogenase 1 alpha subcomplex subunit 4‐like 2 (NDUFA4L2) on myocardial ischemia‐reperfusion (I/R) injury and the underlying molecular mechanism. The hypoxia‐reperfusion (H/R) model was established in vitro using H9c2 cells to simulate I/R injury. NDUFA4L2 and complex I expression levels were detected using RT‐PCR and western blot. The apoptosis of H9C2 cells was evaluated by flow cytometry and the expression of Bax and Bcl‐2. The mitochondrial function were was assessed by ATP concentration, mPTP opening and cytochrome c (cyto C) expression, as well as. Our data indicated that NDUFA4L2 expression was significantly down‐regulated in myocardial H/R injury. Overexpression of NDUFA4L2 led to a dramatic prevention of H/R‐induced apoptosis accompanied by a decrease in the expression of Bax and an increase in the expression of Bcl‐2. Meanwhile, augmentation of NDUFA4L2 dramatically prevented mitochondrial dysfunction caused by H/R, as reflecting reflected in the increased ATP concentration, delayed mPTP opening, as well as down‐regulated cyto C expression. Moreover, complex I activation was heightened and negatively regulated by NDUFA4L2. Silencing complex I conspicuously attenuated cell apoptosis and mitochondrial dysfunction. Taken together, our findings demonstrated that NDUFA4L2 protects against H/R injury by preventing H/R‐induced myocardium apoptosis and mitochondrial dysfunction via the complex I, and may be a potential therapeutic approach for attenuating myocardial I/R injury. This article is protected by copyright. All rights reserved.
    April 21, 2017   doi: 10.1111/1440-1681.12768   open full text
  • The release of pro‐inflammatory cytokines is mediated via MAP‐kinases rather than inflammasome signaling pathway in keratinocytes.
    Thomas Ondet, Béatrice Muscatelli‐Groux, Cédric Coulouarn, Sacha Robert, Thomas Gicquel, Aude Bodin, Vincent Lagente, Jean‐Alexis Grimaud.
    Clinical and Experimental Pharmacology and Physiology. April 20, 2017
    Toll like receptors (TLRs) are expressed in the skin and airway epithelial tissues, which are the most important sites of host‐pathogen interactions. TLRs recognize the three‐dimensional structure of pathogen associated molecules and are thus useful markers of the innate immune response. Here, we investigated the role of lipopolysaccharides (LPS) and monosodium urate (MSU) crystals in the activation of the TLR and Nod Like Receptors (NLR) pathways in human keratinocytes. Analysis of the inflammasome compounds revealed that NLRP3 and TLR4, both of which are components of inflammasome complexes involved in the activation of IL‐1β, were not expressed in keratinocytes. Transcriptomic analysis showed that combination of both MSU and LPS priming do not elicit significant result compare to MSU treatment that induced the expression of TLR2, IL‐6 and IL‐8/CXCL8 in the keratinocyte cell line HaCaT. Furthermore, MSU promoted the phosphorylation of Erk1/2 and MAPK14/p38α MAP kinases. We concluded that MSU stimulates a pro‐inflammatory response in keratinocytes via MAP Kinase pathway to induce production of IL‐8/CXCL8 and TLR2. This article is protected by copyright. All rights reserved.
    April 20, 2017   doi: 10.1111/1440-1681.12765   open full text
  • The ‘rectosigmoid brake’: review of an emerging neuromodulation target for colorectal functional disorders.
    Anthony Y Lin, Phil G. Dinning, Tony Milne, Ian P Bissett, Gregory O'Grady.
    Clinical and Experimental Pharmacology and Physiology. April 16, 2017
    The regulation of gastrointestinal motility encompasses several overlapping mechanisms including highly regulated and coordinated neurohormonal circuits. Various feedback mechanisms or ‘brakes’ have been proposed. While duodenal, jejunal, and ileal brakes are well described, a putative distal colonic brake is less well defined. Despite the high prevalence of colonic motility disorders, there is little knowledge of colonic motility owing to difficulties with organ access and technical difficulties in recording detailed motor patterns along its entire length. The motility of the colon is not under voluntary control. A wide range of motor patterns is seen, with long intervals of intestinal quiescence between them. In addition, the use of traditional manometric catheters to record contractile activity of the colon has been limited by low number of widely spaced sensors, which has resulted in the misinterpretation of colonic motor patterns. The recent advent of high‐resolution (HR) manometry is revolutionising the understanding of gastrointestinal motor patterns. It has now been observed that the most common motor patterns in the colon are repetitive 2‐6 cycles per minute (cpm) propagating events in the distal colon. These motor patterns are prominent soon after a meal, originate most frequently in the rectosigmoid region, and travel in the retrograde direction. The distal prominence and the origin of these motor patterns raise the possibility of them serving as a braking mechanism, or the ‘rectosigmoid brake,’ to limit rectal filling. This review aims to describe what is known about the ‘rectosigmoid brake,’ including its physiological and clinical significance and potential therapeutic applications. This article is protected by copyright. All rights reserved.
    April 16, 2017   doi: 10.1111/1440-1681.12760   open full text
  • Ergogenic properties of metformin in simulated high altitude.
    Rebecca L. Scalzo, Hunter L. Paris, Scott E. Binns, Janelle L. Davis, Joseph W. Beals, Christopher L. Melby, Gary J. Luckasen, Matthew S. Hickey, Benjamin F. Miller, Karyn L. Hamilton, Christopher Bell.
    Clinical and Experimental Pharmacology and Physiology. April 10, 2017
    Metformin augments glucose/glycogen regulation and may acutely promote fatigue resistance during high‐intensity exercise. In hypobaric environments, such as high altitude, the important contribution of carbohydrates to physiological function is accentuated as glucose/glycogen dependence is increased. Because hypoxia/hypobaria decreases insulin sensitivity, replenishing skeletal muscle glycogen in high altitude becomes challenging and subsequent physical performance may be compromised. We hypothesized that in conditions where glycogen repletion was critical to physical outcomes, metformin would attenuate hypoxia‐mediated decrements in exercise performance. On three separate randomly ordered occasions, 13 healthy men performed glycogen‐depleting exercise and ingested a low‐carbohydrate dinner (1200kcals, <10% carbohydrate). The next morning, in either normoxia or hypoxia (FiO2=0.15), they ingested a high‐carbohydrate breakfast (1225kcals, 70% carbohydrate). Placebo (719mg maltodextrin) or metformin (500mg BID) was consumed 3 days prior to each hypoxia visit. Subjects completed a 12.5km cycle ergometer time trial 3.5 hours following breakfast. Hypoxia decreased resting and exercise oxyhemoglobin saturation (P<0.001). Neither hypoxia nor metformin affected the glucose response to breakfast (P=0.977), however, compared with placebo, metformin lowered insulin concentration in hypoxia 45 minutes after breakfast (64.1±6.6 vs. 48.5±7.8 μU/ml; mean±SE; P<0.001). Post‐breakfast, pre‐exercise vastus lateralis glycogen content increased in normoxia (+33%: P=0.025) and in hypoxia with metformin (+81%; P=0.006), but not in hypoxia with placebo (+27%; P=0.167). Hypoxia decreased time trial performance compared with normoxia (P<0.01). This decrement was similar with placebo (+2.6±0.8 min) and metformin (+1.6±0.3 min). These results indicate that metformin promotes glycogen synthesis but not endurance exercise performance in healthy men exposed to simulated high altitude. This article is protected by copyright. All rights reserved.
    April 10, 2017   doi: 10.1111/1440-1681.12761   open full text
  • Tiron ameliorates high glucose‐induced cardiac myocyte apoptosis by PKCδ‐dependent inhibition of osteopontin.
    Ping Jiang, Deling Zhang, Hong Qiu, Xianqi Yi, Yemin Zhang, Yingkang Cao, Bo Zhao, Zhongyuan Xia, Changhua Wang.
    Clinical and Experimental Pharmacology and Physiology. April 10, 2017
    Tiron functions as an effective antioxidant alleviating the intracellular reactive oxygen species (ROS) or the acute toxic metal overload. The previous studies have shown that cardiac myocyte apoptosis can be effectively inhibited by tiron administration in streptozotocin (STZ)‐induced diabetic rats, primary neonatal rat cardiomyocytes (NRVMs), and H9c2 embryonic rat cardiomyocytes. However, the underlying signaling mechanism is ill‐defined. In the present study, we found that tiron supplementation significantly inhibited apoptosis of high glucose (HG)‐treated NRVMs and the left ventricular cardiomyocytes from STZ‐diabetic rat, accompanied with a reduction of osteopontin (OPN) levels as well as an inhibition of PKCδ phosphorylation. OPN knockdown protected NRVMs against HG‐induced cell apoptosis. In addition, genetic inhibition of PKCδ mitigated HG‐stimulated enhancement of intracellular OPN levels in NRVMs. These finding indicate that ROS‐mediated activation of PKCδ upregulated OPN expression, leading to cardiac myocyte apoptosis. Interfering with ROS/PKCδ pathway by antioxidants such as tiron provides an optional therapeutic strategy for treatment and prevention of apoptosis‐related cardiovascular diseases including diabetic cardiomyopathy. This article is protected by copyright. All rights reserved.
    April 10, 2017   doi: 10.1111/1440-1681.12762   open full text
  • Effects of Presynaptic Muscarinic Receptor Blockade on Neuromuscular Transmission as Assessed by the Train‐of‐Four and the Tetanic Fade Response to Rocuronium.
    Yong Beom Kim, Sangseok Lee, Kyeong Cheon Lee, Ha Jung Kim, Young Jin Ro, Hong‐Seuk Yang.
    Clinical and Experimental Pharmacology and Physiology. April 10, 2017
    This study investigated the effect of muscarinic M1 and M2 receptor antagonists on the rocuronium‐induced train of four (TOF) fade and tetanic fade, respectively. Ex‐vivo phrenic nerves/diaphragms were obtained from adult Sprague‐Dawley rats and, stabilized in Krebs buffer, and the nerve‐stimulated muscle TOF fade was observed at 20 s intervals. For the TOF study, phrenic nerve/diaphragms were incubated with pirenzepine (an M1 blocker) at concentrations of 0 nM (control), 10 nM (PZP10), or 100 nM (PZP100). Rocuronium was then administered incrementally until the first twitch tension (T1) had depressed by >95% during TOF stimulation. The mean TOF ratios were compared when the T1 tensions were depressed by 40% to 50%. For the tetanic fade study, 50 Hz/5 s tetani were applied initial, 30 min after administering a loading dose of rocuronium, and 30 min after administering methoctramine (an M2 receptor blocker, loaded at 0 μM (control), 1 μM (MET1), or 10 μM (MET10)). The EC95 of rocuronium was significantly lower in the PZP10 than in the control group. In the PZP10 group, the TOF ratios (TOFRs) at 50s%, T1 depression were significantly lower than those in the control group (p = 0.02). During tetanic stimulation, the tetanic fade was significantly enhanced in the MET10 group compared to the other groups. The study shows that antagonists of muscarinic M1 and M2 receptors affect the rocuronium‐induced neuromuscular block evidenced as shown by the lowering of EC95 and TOFR (M1 antagonist, pirenzepine) or enhancing 50 Hz tetanic fade (M2 antagonist, methoctramine). This article is protected by copyright. All rights reserved.
    April 10, 2017   doi: 10.1111/1440-1681.12763   open full text
  • Compared effects on cerebral oxygenation of ephedrine vs. phenylephrine to treat hypotension during carotid endarterectomy.
    Jugurtha Aliane, Christian Dualé, Nader Guesmi, Charlotte Baud, Eugenio Rosset, Bruno Pereira, Damien Bouvier, Pierre Schoeffler.
    Clinical and Experimental Pharmacology and Physiology. April 04, 2017
    While both ephedrine and phenylephrine are currently used to treat hypotension occurring during carotid endarterectomy (CEA) under general anaesthesia, phenylephrine may have deleterious effects on the cerebral watershed, due to its exclusively vasoconstrictive action. In this controlled, double‐blind randomised trial, we compared the effects of ephedrine and phenylephrine administered in a standardised algorithm to treat the first hypotensive event occurring since induction of anaesthesia until carotid cross‐clamping. The algorithm consisted of 1‐to‐3 boluses of 6 mg of ephedrine or 50 μg of phenylephrine, after a goal‐directed fluid therapy. In case of failure, the treatment switched to the other study drug. Cerebral tissue oxygen saturation (SctO2) was monitored by near infrared spectroscopy (NIRS), and the primary outcome was the restoring effect of SctO2 (ipsilateral to surgery) to baseline values. Secondary postoperative outcomes were: contralateral SctO2, neurological outcomes, and plasma S100B protein measured at discharge from post‐anaesthesia care unit. Ephedrine treatment provided a higher rate of restoration of ipsilateral SctO2 than phenylephrine (93.2% vs. 85.1%, P=0.034); this was also noted for contralateral SctO2 (93.5% vs. 90.7%, P=0.026). The gain in SctO2 on the lowest value during hypotension was also higher under ephedrine than phenylephrine (6.4% vs 4.3% ipsilateral, 5.1% vs. 4% contralateral), but not significantly so. Clinical outcomes were unaffected by the treatment, but S100B protein plasma concentration was higher in the phenylephrine group. To conclude, this pilot trial, focusing on intermediate outcomes, suggests that ephedrine should be preferred to phenylephrine to treat hypotension during CEA. This article is protected by copyright. All rights reserved.
    April 04, 2017   doi: 10.1111/1440-1681.12759   open full text
  • GDF8 inhibits bone formation and promotes bone resorption in mice.
    Yu‐Si Chen, Qi Guo, Li‐Juan Guo, Ting Liu, Xian‐Ping Wu, Zhang‐Yuan Lin, Hong‐Bo He, Tie‐Jian Jiang.
    Clinical and Experimental Pharmacology and Physiology. March 27, 2017
    Growth Differentiation Factor 8 (GDF8), also called myostatin, is a member of the transforming growth factor (TGF)‐β super‐family. As a negative regulator of skeletal muscle growth, GDF8 is also associated with bone metabolism. However, the function of GDF8 in bone metabolism is not fully understood. Our study aimed to investigate the role of GDF8 in bone metabolism, both in vitro and in vivo. Our results showed that GDF8 had a negative regulatory effect on primary mouse osteoblasts, and promoted receptor activator of nuclear factor κB ligand (RANKL)‐induced osteoclastogenesis in vitro. Intraperitoneal injection of recombinant GDF8 repressed bone formation and accelerated bone resorption in mice. Furthermore, treatment of aged mice with a GDF8 neutralizing antibody stimulated new bone formation and prevented bone resorption. Thus, our study showed that GDF8 plays a significant regulatory role in bone formation and bone resorption, thus providing a potential therapeutic pathway for osteoporosis.
    March 27, 2017   doi: 10.1111/1440-1681.12728   open full text
  • The selective serotonin reuptake inhibitor dapoxetine inhibits voltage‐dependent K+ channels in rabbit coronary arterial smooth muscle cells.
    Han Sol Kim, Hongliang Li, Hye Won Kim, Sung Eun Shin, Won‐Kyo Jung, Kwon‐Soo Ha, Eun‐Taek Han, Seok‐Ho Hong, Amy L Firth, Il‐Whan Choi, Won Sun Park.
    Clinical and Experimental Pharmacology and Physiology. March 27, 2017
    We investigated the inhibitory effect of dapoxetine, a selective serotonin reuptake inhibitor (SSRI), on voltage‐dependent K+ (Kv) channels using native smooth muscle cells from rabbit coronary arteries. Dapoxetine inhibited Kv channel currents in a concentration‐dependent manner, with an IC50 value of 2.68±0.94 μmol/L and a slope value (Hill coefficient) of 0.63±0.11. Application of 10 μmol/L dapoxetine accelerated the rate of inactivation of Kv currents. Although dapoxetine did not modify current activation kinetics, it caused a significant negative shift in the inactivation curves. Application of train step (1 or 2 Hz) progressively increased the inhibitory effect of dapoxetine on Kv channels. In addition, the recovery time constant was extended in its presence, suggesting that the longer recovery time constant from inactivation underlies a use‐dependent inhibition of the channel. From these results, we conclude that dapoxetine inhibits Kv channels in a dose‐, time‐, use‐, and state (open)‐dependent manner, independent of serotonin reuptake inhibition.
    March 27, 2017   doi: 10.1111/1440-1681.12723   open full text
  • Bromocriptine treatment at the end of lactation prevents hyperphagia, higher visceral fat and liver triglycerides in early‐weaned rats at adulthood.
    Nayara Peixoto‐Silva, Egberto G Moura, Janaine C Carvalho, Jéssica L Nobre, Fernanda T Quitete, Cintia R Pinheiro, Ana Paula Santos‐Silva, Elaine Oliveira, Patricia C Lisboa.
    Clinical and Experimental Pharmacology and Physiology. March 27, 2017
    Non‐pharmacological early weaning (NPEW) leads offspring to obesity, higher liver oxidative stress and microsteatosis in adulthood. Pharmacological EW (PEW) by maternal treatment with bromocriptine (BRO) causes obesity in the adult progeny but precludes hepatic injury. To test the hypothesis that BRO prevents the deleterious changes of NPEW, we injected BRO into the pups from the NPEW model in late lactation. Lactating rats were divided into two groups: dams with an adhesive bandage around the body to prevent breastfeeding on the last 3 days of lactation and dams whose pups had free suckling (C). Offspring from both groups were subdivided into two groups: pups treated with BRO (intraperitoneal (i.p.) 4 mg/kg per day) on the last 3 days of lactation (NPEW/BRO and C/BRO) or pups treated with the vehicle (NPEW and C). At PN120, offspring were challenged with a high fat diet (HFD), and food intake was recorded after 30 minutes and 12 hours. Rats were killed at PN120 and PN200. At PN120, adipocyte size was greater in the NPEW group but was normal in the NPEW/BRO group. At PN200, the NPEW group presented hyperphagia, higher adiposity, adipocyte hypertrophy, hyperleptinaemia, glucose intolerance and increased hepatic triglycerides. These parameters were normalized in the NPEW/BRO group. In the feeding test, BRO groups showed lower HFD intake at 30 minutes than did their controls; however, at 12 hours, the NPEW group ate more HFD. The treatment with BRO can preclude some deleterious effects of the NPEW model, which prevented the development of overweight and its comorbidities.
    March 27, 2017   doi: 10.1111/1440-1681.12724   open full text
  • Prognostic value of the MicroRNA‐29 family in multiple human cancers: A meta‐analysis and systematic review.
    Yan Qi, Yalan Huang, Lijuan Pang, Wenyi Gu, Ning Wang, Jianming Hu, Xiaobin Cui, Jun Zhang, Jin Zhao, Chunxia Liu, Wenjie Zhang, Hong Zou, Feng Li.
    Clinical and Experimental Pharmacology and Physiology. March 27, 2017
    MicroRNAs (miRNAs) in cancer development have attracted much attention in recent years. miR‐29 is known to critically affect cancer progression by functioning as a tumor suppressor. However, it may also act as an oncogene under certain situations. The prognostic value of the miR‐29 family in cancer progression is still under debate and reported results are inconsistent. Therefore, we reported here a meta‐analysis and systematic review to analyze the prognostic role of the miR‐29 family in cancer. We screened 20 published studies and calculated pooled hazard ratios (HRs) and corresponding 95% confidence intervals (CIs) for overall survival (OS) or disease‐free survival/recurrence‐free survival (DFS/RFS). Our results showed that a low or absent expression of miR‐29 family was significantly associated with poor OS (HR, 1.57; 95%CI, 1.18‐2.08), and inferior to 5‐year DFS/RFS (HR, 1.89; 95%CI, 1.47‐2.44). Analysis of individual miR‐29 subtypes indicated that the low expression of miR‐29a/b/c subtypes correlated with poor 5‐year OS (miR‐29a: HR, 1.99; 95%CI, 1.41‐2.80; miR‐29b: HR, 1.60; 95%CI, 1.18‐2.17; miR‐29c: HR, 1.69; 95%CI, 1.00‐2.86), as well as poor 5‐year DFS/RFS (miR‐29b: HR, 1.70; 95%CI, 1.27‐2.27). Ethnicity analysis demonstrated Asian patients with low expression of miR‐29 were significantly correlated with poor OS (HR, 1.61; 95%CI, 1.16‐2.23) and 5‐year DFS/RFS (HR, 2.03; 95%CI, 1.50‐2.74). Taken together, our analysis indicates that the low expression of miR‐29 is associated with aggressiveness and poor prognosis of malignant neoplasms. More importantly, miR‐29 might serve as a key biomarker for predicting the recurrence and progression of human cancers.
    March 27, 2017   doi: 10.1111/1440-1681.12726   open full text
  • Effects of electrical lesion of basolateral amygdala nucleus on rat anxiety‐like behaviour under acute, sub‐chronic, and chronic stresses.
    Hoda Ranjbar, Maryam Radahmadi, Parham Reisi, Hojjatallah Alaei.
    Clinical and Experimental Pharmacology and Physiology. March 27, 2017
    Stress contributes, as a risk factor, to such psychological disorders as anxiety. The effects of electrical lesions in the basolateral amygdala nucleus (BLA) were investigated on the locomotor activity and anxiety‐like behaviour in different stress durations. For this purpose, rats were randomly allocated to control, sham, and experimental groups, the latter including groups with and without BLA nucleus subjected to acute, sub‐chronic, and chronic stress conditions for 1, 7, and 21 days, respectively, applied 6 h/d. The induced anxiety behaviour was evaluated using the open field test (OFT) while other variables were measured. Findings revealed that sub‐chronic stress led to significantly reduced (P<.05) anxiety behaviours as measured by entries into and the time spent in the centre area while it also led to significant impairments in exploratory and locomotor activities, indicating intensified anxiety‐like behaviour. BLA lesion affected rat behaviour differently such that it significantly (P<.05) decreased fear under sub‐chronic and chronic stress conditions as evidenced by the subjects’ greater tendency to enter the centre area in the open field test and their increased number of rearing events (P<.01). However, BLA lesion led to no significant decrease in the locomotor activity of subjects exposed to sub‐chronic or chronic stress conditions as compared with those in similar groups but without BLA lesion. Finally, BLA lesion was found not only to decrease significantly (P<.01) adrenal gland and body weights, particularly under sub‐chronic stress, but also to play a critical role in modulating adrenal functions by decreasing adrenal gland weight, and thereby reducing depression‐like symptoms.
    March 27, 2017   doi: 10.1111/1440-1681.12727   open full text
  • Acute cardiac support with intravenous milrinone promotes recovery from early brain injury in a murine model of severe subarachnoid haemorrhage.
    Tomoko Mutoh, Tatsushi Mutoh, Kazuhiro Nakamura, Yukiko Yamamoto, Yoshiharu Tsuru, Hirokazu Tsubone, Tatsuya Ishikawa, Yasuyuki Taki.
    Clinical and Experimental Pharmacology and Physiology. March 27, 2017
    Early brain injury/ischaemia (EBI) is a serious complication early after subarachnoid haemorrhage (SAH) that contributes to development of delayed cerebral ischaemia (DCI). This study aimed to determine the role of inotropic cardiac support using milrinone (MIL) on restoring acute cerebral hypoperfusion attributable to EBI and improving outcomes after experimental SAH. Forty‐three male C57BL/6 mice were assigned to either sham surgery (SAH‐sham), SAH induced by endovascular perforation plus postconditioning with 2% isoflurane (Control), or SAH plus isoflurane combined with MIL with and without hypoxia‐inducible factor inhibitor (HIF‐I) pretreatment. Cardiac output (CO) during intravenous MIL infusion (0.25‐0.75 μg/kg/min) between 1.5 and 2.5 hours after SAH induction was monitored with Doppler echocardiography. Magnetic resonance imaging (MRI)‐continuous arterial spin labelling was used for quantitative cerebral blood flow (CBF) measurements. Neurobehavioral function was assessed daily by neurological score and open field test. DCI was analyzed 3 days later by determining infarction on MRI. Mild reduction of cardiac output (CO) and global cerebral blood flow (CBF) depression were notable early after SAH. MIL increased CO in a dose‐dependent manner (P<.001), which was accompanied by improved hypoperfusion, incidence of DCI and functional recovery than Control (P<.05). The neuroprotective effects afforded by MIL or Control were attenuated by hypoxia‐inducible factor (HIF) inhibition (P<.05). These results suggest that MIL improves acute hypoperfusion by its inotropic effect, leading to neurobehavioral improvement in mice after severe SAH, in which HIF may be acting as a critical mediator.
    March 27, 2017   doi: 10.1111/1440-1681.12718   open full text
  • Regulatory B cell is critical in bone union process through suppressing proinflammatory cytokines and stimulating Foxp3 in Treg cells.
    Guojing Sun, Yicun Wang, Yunfan Ti, Jun Wang, Jianning Zhao, Hongbo Qian.
    Clinical and Experimental Pharmacology and Physiology. March 27, 2017
    Bone fractures may result in delayed union (DU) or non‐union (NU) in some patients. Evidence suggests that the skewing of the immune system toward the proinflammatory type is a contributing factor. Because B cells were previously found to infiltrate the fracture healing site at abundant levels, we examined the regulatory B cells (Bregs) in DU/NU patients. In bone fracture patients with normal healing, the frequency of interleukin (IL)‐10‐expressing B cells was significantly upregulated in the early healing process (6 weeks post‐surgery) and was downregulated later on (18 weeks post‐surgery), whereas in DU/NU patients, the early upregulation of IL‐10‐expressing B cells was missing. The majority of IL‐10‐expressing B cells were concentrated in the IgM+CD27+ fraction in both controls and patients. IgM+CD27+ B cells effectively suppressed interferon gamma (IFN‐γ), tumor necrosis factor alpha (TNF‐α), and IL‐2 expression from CD4+ T cells, as well as IFN‐γ and TNF‐α expression from CD8+ T cells. The IgM+CD27+ B cell‐mediated suppression was restricted to the sample from the early healing time point in controls, as the IgM+CD27+ B cells from normal healing patients later on or from DU/NU patients did not present significant regulatory function. In addition, culturing of CD4+CD25+ Tregs with IgM+CD27+ B cells from controls at early healing time point resulted in higher Foxp3 expression, a function absent in controls at later time point, or in DU/NU patients. In conclusion, our results support a role of B cell‐mediated regulation early during the bone healing process.
    March 27, 2017   doi: 10.1111/1440-1681.12719   open full text
  • Effects of miRNAs on myocardial apoptosis by modulating mitochondria related proteins.
    Yanfang Zhao, Murugavel Ponnusamy, Yanhan Dong, Lei Zhang, Kun Wang, Peifeng Li.
    Clinical and Experimental Pharmacology and Physiology. March 27, 2017
    Myocardial apoptosis play a vital role in pathogenesis of cardiovascular diseases. The intrinsic pathway of apoptosis (mitochondrial apoptosis pathway) and abnormal mitochondrial fission and fusion have a detrimental effect on cells under a variety of intracellular stresses including hypoxia, oxidative stress, drug toxicity or DNA damage and contributes to the development of heart failure (HF), myocardial infarction (MI), diabetic cardiomyopathy and ischaemia/reperfusion injury (I/R). MicroRNAs (miRNAs) are endogenous short non‐coding RNAs, which target 3′‐untranslated region of mRNA to switch off gene expression. They play crucial roles in regulating complicated cardiac signalling and transcriptional events during cardiac development as well as in diseased condition. In this review, we summarize the molecular mechanism of mitochondrial apoptosis in cardiac cells and influence of miRNAs on them. MiRNAs regulate cardiac mitochondrial apoptosis by exert their effects on mitochondrial fission and fusion, reactive oxygen species (ROS) generation and Ca2+ homeostasis, Bcl‐2 family members, and other mitochondrial function proteins. This advancement in understanding mechanism of cardiac cells death provides us new therapy targets for cardiovascular diseases associated with mitochondrial dysfunctions.
    March 27, 2017   doi: 10.1111/1440-1681.12720   open full text
  • Renoprotective effect of Hypericum perforatum against diabetic nephropathy in rats: Insights in the underlying mechanisms.
    Dalia M. Abd El Motteleb, Dalia I. Abd El Aleem.
    Clinical and Experimental Pharmacology and Physiology. March 27, 2017
    Oxidative stress and inflammation play a key role in the initiation and progression of diabetic nephropathy (DN). The present study aimed to investigate the possible protective effect of hypericum perforatum (HP) against DN. Rats were allocated into six groups: control, received normal saline; diabetic untreated (DM), received single dose of streptozotocin (STZ) after injection of nicotinamide (NA); gliclazide, received STZ,NA + gliclazide (10 mg/kg); DM + HP50, DM + HP100, DM + HP200, received STZ,NA and HP 50, 100, 200 mg/kg, respectively. Gliclazide and HP were administered daily via gavage for 8 weeks. Serum glucose, insulin, kidney function and histopathological picture were assessed. Furthermore, oxidative/nitrosative stress, inflammatory cytokines, apoptotic and fibrotic markers were measured. Diabetic untreated group showed increase in serum glucose, urea, creatinine with albuminurea. Renal expression of protein for nuclear factor kappa‐B (NF‐кB), renal expression of inducible nitric oxide synthase (iNOS), cyclooxygenase II (COXII), collagen IV, fibronectin were elevated. Malondialdehyde (MDA), nitric oxide (NO), tumour necrosis factor‐α (TNF‐α), interleukin‐1β (IL‐1β), intracellular adhesion molecule (ICAM‐1), monocellular chemoattractant protein‐1 (MCP‐1), tumour growth factor‐ β (TGF‐β), caspase‐3 and cytochrome c contents were also increased consequently with decline of serum insulin, expression of peroxisome proliferator‐activated receptor (PPARγ), renal reduced glutathione (GSH) content and superoxide dismutase (SOD) activity. Treatment with either gliclazide or HP mitigated the deleterious effects of STZ on the tested parameters. These findings indicate for the first time that HP may have a renoprotective effect against DN through reduction of oxidative/nitrosative stress, enhancement of antioxidant defense mechanisms, decline of inflammatory cytokines, antifibrotic, antiapoptotic and blood glucose lowering properties.
    March 27, 2017   doi: 10.1111/1440-1681.12729   open full text
  • On the use of one‐sided statistical tests in biomedical research.
    Ricardo Murphy.
    Clinical and Experimental Pharmacology and Physiology. March 21, 2017
    There is a tendency to automatically use two‐sided tests to assess the statistical significance of experimental results. Yet if a theory predicts the direction of an experimental outcome, or if for some practical (e.g. clinical) reason an outcome in that direction is the only one of interest, then it makes sense to use a one‐sided test. The use of a two‐sided test in these situations will lead to too many false negatives. Consequently treatment effects that corroborate a theory or that are of practical importance may be missed. This problem becomes particularly acute in the case of borderline results. Following a nonsignificant one‐sided test, the possibility of an effect in the direction opposite to that predicted or required can be assessed in an exploratory fashion by computing the odds in favour of such an effect. Anyone is then at liberty to pursue this possibility as they see fit. The question of whether to use a one‐sided or two‐sided statistical test should always be decided on logical grounds not statistical ones, and suspicions regarding the motives of the investigator(s) should be disregarded. On the other hand, this choice can be avoided altogether by assuming that a treatment always has some effect (however small) and then computing the strength of the evidence in favour of the observed or predicted/required effect (i.e. 1‐P, where P is the one‐sided significance level of the test). With this approach one‐sided and two‐sided tests yield identical results, and so there is effectively only one type of test. This article is protected by copyright. All rights reserved.
    March 21, 2017   doi: 10.1111/1440-1681.12754   open full text
  • STAT3 down‐regulation induces mitochondria‐dependent G2/M cell cycle arrest and apoptosis in oesophageal carcinoma cells.
    Dan Shao, Jie Ma, Chao Zhou, Jia‐Nan Zhao, Lu‐Lu Li, Tong‐Jian Zhao, Xi‐Lei Ai, Ping Jiao.
    Clinical and Experimental Pharmacology and Physiology. February 14, 2017
    STAT3 is persistently activated in a wide variety of human tumours, and aberrant STAT3 activity promotes tumour growth, invasion and metastasis. To explore STAT3 down‐regulation in human oesophageal cancer cells, cell proliferation, apoptosis and mitochondrial mechanisms were explored in oesophageal carcinoma TE1 cell cultures. We demonstrate for the first time that STAT3 down‐regulation by RNAi is sufficient to inhibit oesophageal cancer cell proliferation inducing cell apoptosis. Further, we demonstrate that mitochondrial transmembrane potential is impaired thereby leading to collapsed mitochondrial membrane potential, abnormal mitochondrial membrane depolarization, nuclear DNA fragmentation and cell cycle G2/M arrest under the conditions of STAT3 down‐regulation. Thus, our results suggest that STAT3 inhibition is a valid approach to induce oesophageal carcinoma cell mitochondrial‐dependent apoptosis in therapeutic strategies against oesophageal cancers.
    February 14, 2017   doi: 10.1111/1440-1681.12708   open full text
  • Isoquercitrin protects against pulmonary hypertension via inhibiting PASMCs proliferation.
    Yongtao Zhang, Yuqian Cui, Wei Deng, Hao Wang, Weidong Qin, Chengmin Huang, Chen Li, Jianning Zhang, Yuan Guo, Dawei Wu, Haipeng Guo.
    Clinical and Experimental Pharmacology and Physiology. February 14, 2017
    Pulmonary vascular remodelling is a common feature among the heterogeneous disorders that cause pulmonary arterial hypertension (PAH), and pulmonary arterial smooth muscle cells (PASMCs) proliferation impact the long‐term prognosis of the patient. Isoquercitrin (IQC) is a flavonoid with anti‐oxidative, anti‐inflammatory and anti‐proliferative activations. This study aimed to investigate whether IQC could prevent PASMCs proliferation and vascular remodelling in monocrotaline (MCT) induced PAH. Male Wistar rats were administered with Vehicle or 0.1% IQC maintain feed after MCT (40 mg/kg) injection. Haemodynamic changes, right ventricular hypertrophy and lung morphological features were assessed 3 weeks later. MCT‐induced PAH, pulmonary vascular remodelling and PASMCs proliferation in Vehicle‐treated rats. IQC reduced the right ventricle systolic pressure (RVSP), the ratio of RV/LV+S and the RV hypertrophy. IQC significantly alleviated the expression of proliferating cell nuclear antigen (PCNA), smooth muscle α‐actin (α‐SMA), and the percentage of fully muscularized small arterioles. In vitro studies, PASMCs were pretreated with IQC and stimulated with platelet‐derived growth factor (PDGF)‐BB (20 ng/mL). IQC suppressed PDGF‐BB‐induced PASMCs proliferation and caused G0/G1 phase cell cycle arrest. IQC downregulated the expression of Cyclin D1 and CDK4 as well as inhibited p27Kip1 degradation. Meanwhile, IQC negatively modulated PDGF‐BB‐induced phosphorylation of PDGF‐Rβ, Akt/GSK3β and ERK1/2. IQC ameliorated MCT‐induced pulmonary vascular remodelling via suppressing PASMCs proliferation and blocking PDGF‐Rβ signalling pathway.
    February 14, 2017   doi: 10.1111/1440-1681.12705   open full text
  • Physical exercise remodels visceral adipose tissue and mitochondrial lipid metabolism in rats fed a high‐fat diet.
    Sílvia Rocha‐Rodrigues, Amaia Rodríguez, Sara Becerril, Beatriz Ramírez, Inês O Gonçalves, Jorge Beleza, Gema Frühbeck, António Ascensão, José Magalhães.
    Clinical and Experimental Pharmacology and Physiology. February 14, 2017
    We aimed to investigate the effects of two physical exercise models, voluntary physical activity (VPA) and endurance training (ET) as preventive and therapeutic strategies, respectively, on lipid accumulation regulators and mitochondrial content in VAT of rats fed a high‐fat diet (HFD). Sprague‐Dawley rats (6 weeks old, n=60) were assigned into sedentary and VPA groups fed isoenergetic diets: standard (S, 35 kcal% fat) or HFD (71 kcal% fat). The VPA groups had free access to wheel running during the entire protocol. After 9 weeks, half of the sedentary animals were exercised on a treadmill while maintaining the dietary treatments. The HFD induced no changes in plasma non‐esterified fatty acids (NEFA) and glycerol levels and decreased oxidative phosphorylation (OXPHOS) subunit IV and increased truncated/full‐length sterol regulatory element‐binding transcription factor 1c (SREBP1c) ratio in epididymal white adipose tissue (eWAT). VPA decreased plasma glycerol levels, aquaglyceroporin 7 (AQP7) and increased subunit I of cytochrome c oxidase (COX) protein, in standard diet fed animals. Eight weeks of ET decreased body weight, visceral adiposity and adipocyte size and plasma NEFA and glycerol levels, as well as AQP7 protein expression in eWAT. ET increased fatty acid translocase (FAT/CD36), mitochondrial content of complexes IV and V subunits, mitochondrial biogenesis and dynamic (mitofusins and optic atrophy 1)‐related proteins. Moreover, lipogenesis‐related markers (SREBP1c and acetyl CoA carboxylase) were reduced after 8 weeks of ET. In conclusion, ET‐induced alterations reflect a positive effect on mitochondrial function and the overall VAT metabolism of HFD‐induced obese rats.
    February 14, 2017   doi: 10.1111/1440-1681.12706   open full text
  • Alleviative effect of ciliary neurotrophic factor analogue on high fat‐induced hepatic steatosis is partially independent of the central regulation.
    Ming‐Xia Cui, Li‐Ning Yang, Xiao‐Xia Wang, Lei Wang, Rui‐Lian Li, Wei Han, Yong‐Jie Wu.
    Clinical and Experimental Pharmacology and Physiology. February 14, 2017
    Ciliary neurotrophic factor (CNTF) analogues were reported to ameliorate fatty liver in db/db or high‐fat diet‐fed mice. It is generally thought that CNTF exerts its actions centrally. The aim of this study was to investigate whether peripheral effects of CNTF analogues are involved in the therapeutic effect on high fat‐induced hepatic steatosis. The rat model of fatty liver was induced by a high‐fat diet (HFD) for 12 weeks. In the next 2 weeks, rats were fed the HFD along with subcutaneous injection of vehicle or mutant recombinant human CNTF (rhmCNTF 0.05‐0.2 mg/kg per day). Steatotic HepG2 cells were induced by 50% fetal bovine serum (FBS) for 48 hours, and then treated with rhmCNTF for 24 hours. The results showed that after rhmCNTF treatment, hepatic triglyceride (TG) accumulation was attenuated both in vivo and in vitro. RhmCNTF increased protein expression of CPT‐1 and PPARα, and decreased SREBP‐1c, FAS and SCD‐1 in steatotic HepG2 cells. But the production of nitric oxide and 8‐isoPGF2α in steatotic HepG2 cells was not affected by rhmCNTF. These results suggest that rhmCNTF has a peripheral effect that alleviates fat‐induced hepatic steatosis.
    February 14, 2017   doi: 10.1111/1440-1681.12709   open full text
  • Effects of unilatral‐ and bilateral inhibition of rostral ventral tegmental area and central nucleus of amygdala on morphine‐induced place conditioning in male Wistar rat.
    Zahra Mohammadian, Hedayat Sahraei, Gholam Hossein Meftahi, Hengameh Ali‐Beik.
    Clinical and Experimental Pharmacology and Physiology. February 14, 2017
    The rostral ventral tegmental area (VTAR) and central nucleus of amygdala (CeA) are considered the main regions for induction of psychological dependence on abused drugs, such as morphine. The main aim of this study was to investigate the transient inhibition of each right and left side as well as both sides of the VTAR and the CeA by lidocaine (2%) on morphine reward properties using the conditioned place preference (CPP) method. Male Wistar rats (250±20 g) 7 days after recovery from surgery and cannulation were conditioned to morphine (7.5 mg/kg) in CPP apparatus. Five minutes before morphine injection in conditioning phase, lidocaine was administered either uni‐ or bilaterally into the VTAR (0.25 μL/site) or CeA (0.5 μL/site). The results revealed that lidocaine administration into the left side, but not the right side of the VTAR and the CeA reduced morphine CPP significantly. The reduction was potentiated when lidocaine was injected into both sides of the VTAR and the CeA. The number of compartment crossings was reduced when lidocaine was injected into both sides of the VTAR and the CeA as well as the left side. Rearing was reduced when lidocaine was injected into the right, but not the left side of the VTAR. Sniffing and rearing increased when animals received lidocaine in the right side and reduced in the group that received lidocaine in the left side of the CeA. It was concluded that the right and the left side of VTAR and the CeA play different roles in morphine‐induced activity and reward.
    February 14, 2017   doi: 10.1111/1440-1681.12715   open full text
  • Pitavastatin suppresses hyperglycaemia‐induced podocyte injury via bone morphogenetic protein‐7 preservation.
    Makoto Ohigashi, Miyuki Kobara, Tamotsu Takahashi, Hiroe Toba, Takehiko Wada, Tetsuo Nakata.
    Clinical and Experimental Pharmacology and Physiology. February 14, 2017
    Podocytes form the essential components of the glomerular filtration barrier and play a critical role in diabetic nephropathy. Recent evidence suggests that HMG‐CoA reductase inhibitors (statins) exert renoprotective effects. We investigated whether pitavastatin directly suppresses hyperglycaemia‐induced podocyte injury using cultured podocytes and, if so, the mechanism of the beneficial effects. Cultured podocytes were exposed to media containing normal (NG; 5 mmol/L) or high (HG; 25 mmol/L) glucose for 1 week. HG increased the lethal injury of podocytes and disruption of F‐actin fibers, and reduced the mRNA expression of novel podocyte markers, synaptopodin and Wilms tumor‐1 (WT‐1), in association with decreased bone morphogenetic protein‐7 (BMP‐7) expression. Pitavastatin (100 nmol/L) reduced podocyte injury and restored the mRNA expression of synaptopodin and WT1; however, these protective effects were abolished by BMP‐7 siRNA. Additionally, pitavastatin suppressed HG‐induced Rho kinase activation, as assessed by the phosphorylation level of myosin phosphatase targeting subunit 1 (MYTP1), and C3 exotoxin, a Rho inhibitor, mimicked the effect of pitavastatin on BMP‐7 preservation. Pitavastatin attenuates hyperglycaemia‐induced podocyte injury via Rho‐Rho kinase‐dependent BMP‐7 preservation.
    February 14, 2017   doi: 10.1111/1440-1681.12716   open full text
  • Strains of Group B streptococci from septic patients induce platelet activation via Toll‐like Receptor 2.
    Xiaoyan Liu, Hongyun Liu, Xianming Luo, Ping Zhang, Yanmin Gao, Shuangfeng Xie, Kang Xu, Jianxing Chang, Liping Ma.
    Clinical and Experimental Pharmacology and Physiology. February 14, 2017
    Group B Streptococcus (GBS) causes life‐threatening bacterial sepsis, especially in newborns and pregnant women. Patients suffering from sepsis often display low platelet counts, characterized by thrombocytopenia, because of platelet activation. In the present study, the roles of six GBS strains from septic patients in platelet aggregation, as well as the underlying mechanisms, were investigated. Incubation of platelets with three of the strains induced platelet aggregation, increased the secretion of cellular adhesin molecule CD62P and activation of GPIIb/IIIa. Furthermore, the GBS strains that induced platelet activation also caused an increase in the expression of Toll‐like receptor (TLR) 2 in platelets. Pre‐incubation of platelets with anti‐TLR2 monoclonal antibody, but not anti‐TLR4 monoclonal antibody, inhibited these functional responses induced by GBS. TLR2 stimulation also activated the phosphoinositide 3‐kinase (PI3‐K)/Akt signalling pathway in platelets, and inhibition of PI3‐K significantly reduced GBS‐induced platelet responses. Our results indicate that three of the GBS strains from the septic patients can trigger platelet activation by interacting with platelets, which involves the elevation of platelet TLR2 expression.
    February 14, 2017   doi: 10.1111/1440-1681.12707   open full text
  • Pravastatin attenuates the action of the ETS domain‐containing protein ELK1 to prevent atherosclerosis in apolipoprotein E‐knockout mice via modulation of extracellular signal‐regulated kinase 1/2 signal pathway.
    Wei Yan, Dan Li, Xiaoxu Zhou.
    Clinical and Experimental Pharmacology and Physiology. February 14, 2017
    Oxidative stress plays an important role in atherosclerosis, a vascular disease with high morbidity and mortality. The ETS domain‐containing protein ELK1 is an oxidative stress‐sensitive factor modulated by the extracellular signal‐regulated kinase (ERK) 1/2 pathway. However, the role of ELK1 in the prevention of atherosclerosis by pravastatin remains unclear. In the present study, male apolipoprotein E‐knockout (apoE−/−) mice fed a diet containing 1.25% cholesterol (w/w) were divided into two groups, one treated with pravastatin (80 mg/kg, 2‐2.4 mg/mouse per day) for 8 weeks and the other not. Male C57BL/6J mice fed with a normal diet were used as a control group. Human umbilical vein endothelial cells (HUVEC) were cultured and treated with pravastatin (10 μmol/L) for 18 hours before testing for the presence or absence of 100 μmol/L H2O2 (24 hours). Examination of pathological sections from mice aortas revealed that pravastatin treatment almost prevented atherosclerotic plaque formation. Pravastatin also inhibited increases in serum and aortic levels of oxidized low‐density lipoprotein and aortic malondialdehyde levels and decreases in aortic reduced glutathione, and the activities of superoxide dismutase, catalase and glutathione peroxidase. H2O2‐induced increases in reactive oxygen species in HUVECs were reversed by pravastatin by 48%. Pravastatin blocked the phosphorylation of ELK1 and ERK1/2 proteins and reduced mRNA levels of early growth response 1, a known atherogenic transcription factor upregulated by the ROS/ERK/ELK1 pathway, in mice. In conclusion, pravastatin attenuates the action of ELK1 induced by oxidative stress to prevent atherosclerosis, which is dependent partly on modulation of ERK1/2 signalling.
    February 14, 2017   doi: 10.1111/1440-1681.12710   open full text
  • Dexmedetomidine preconditioning for myocardial protection in ischaemia‐reperfusion injury in rats by downregulation of the high mobility group box 1‐toll‐like receptor 4‐nuclear factor κB signalling pathway.
    Yu‐fan Yang, Ke Peng, Hong Liu, Xiao‐wen Meng, Jing‐jing Zhang, Fu‐hai Ji.
    Clinical and Experimental Pharmacology and Physiology. February 14, 2017
    Pharmacological preconditioning reduces myocardial infarct size in ischaemia‐reperfusion (I‐R) injury. Dexmedetomidine, a selective α2‐adrenoceptor agonist, has a proven cardioprotective effect when administered prior to I‐R, although the underlying mechanisms for this effect are not fully understood. We evaluated whether dexmedetomidine preconditioning could induce a myocardio‐protective effect against I‐R injury by inhibiting associated inflammatory processes through downregulation of the high mobility group box 1 (HMGB1)‐toll‐like receptor 4 (TLR4)‐nuclear factor κB (NF‐κB) signalling pathway. Seventy rats were randomly assigned to seven groups: a control and six test groups, involving I‐R for 30 and 120 minutes, respectively, in isolated rat hearts and different pretreatment protocols with dexmedetomidine (10 nmol/L) as well as yohimbine (1 μmol/L) and recombinant HMGB1 peptide (rHMGB1; 20 μg/L), alone or in combination with dexmedetomidine. Cardiac function was recorded; myocardial HMGB1, TLR4, and NF‐κB activities and levels of tumour necrosis factor‐α (TNF‐α) and interleukin‐6 (IL‐6) were measured as were lactate dehydrogenase (LDH) and creatine kinase (CK) in coronary outflow. Dexmedetomidine preconditioning significantly improved cardiac function (P<.05), downregulated the expression of HMGB1‐TLR4‐NF‐κB, reduced levels of TNF‐α and IL‐6 in isolated ventricles during I‐R injury, and significantly reduced CK and LDH levels in coronary outflow (P<.05). All of these effects were partially reversed by yohimbine (P<.05) or rHMGB1 (P<.05). Dexmedetomidine preconditioning alleviated myocardial I‐R injury in rats through inhibition of inflammatory processes associated with downregulation of the HMGB1‐TLR4‐NF‐κB signalling pathway via activation at α2‐adrenergic receptors.
    February 14, 2017   doi: 10.1111/1440-1681.12711   open full text
  • Sex differences in ischaemia/reperfusion‐induced acute kidney injury depends on the degradation of noradrenaline by monoamine oxidase.
    Ryosuke Tanaka, Maki Yazawa, Yuri Morikawa, Hidenobu Tsutsui, Mamoru Ohkita, Tokihito Yukimura, Yasuo Matsumura.
    Clinical and Experimental Pharmacology and Physiology. February 14, 2017
    Ischaemic acute kidney injury (AKI) is a leading killer of both sexes; however, resistance to this injury is higher among women than men. We found that renal venous noradrenaline (NAd) overflow after reperfusion played important roles in the development of ischaemic AKI, and that the attenuation of AKI observed in female rats may be dependent on depressing the renal sympathetic nervous system with endogenous oestrogen. In the present study, we used male and female Sprague‐Dawley rats to investigate whether sex differences in the pathogenesis of ischaemic AKI are related to the degradation of NAd by monoamine oxidase (MAO) in the kidney. Ischaemic AKI was achieved by clamping the left renal artery and vein for 45 minutes followed by reperfusion 2 weeks after contralateral nephrectomy. Renal injury was more severe in male rats than in female rats and renal venous plasma NAd levels after reperfusion were markedly elevated in males, but not in females. These sex differences were eliminated by a treatment with isatin, a non‐selective MAO inhibitor, and moclobemide, a selective MAOA inhibitor, but not by selegiline, a selective MAOB inhibitor. Ischaemia decreased the mRNA expression levels of both MAOs in the kidney 1 day after reperfusion; however, MAOA mRNA expression levels were higher in female rats than in male rats. These results suggest that the degradation of NAd by MAOA in the kidney contributes to sex differences in the pathogenesis of ischaemia/reperfusion‐induced AKI.
    February 14, 2017   doi: 10.1111/1440-1681.12713   open full text
  • Antihypertensive methyldopa, labetalol, hydralazine, and clonidine reversed tumour necrosis factor‐α inhibited endothelial nitric oxide synthase expression in endothelial‐trophoblast cellular networks.
    Bei Xu, Gabriele Bobek, Angela Makris, Annemarie Hennessy.
    Clinical and Experimental Pharmacology and Physiology. February 14, 2017
    Medications used to control hypertension in pregnancy also improve trophoblast and endothelial cellular interaction in vitro. Tumour necrosis factor‐α (TNF‐α) inhibits trophoblast and endothelial cellular interactions and simultaneously decreases endothelial nitric oxide synthase (eNOS) expression. This study investigated whether antihypertensive medications improved these cellular interactions by modulating eNOS and inducible nitric oxide synthase (iNOS) expression. Human uterine myometrial microvascular endothelial cells (UtMVECs) were pre‐incubated with (or without) low dose TNF‐α (0.5 ng/mL) or TNF‐α plus soluble fms‐like tyrosine kinase‐1 (sFlt‐1) (100 ng/mL). The endothelial cells were cultured on Matrigel. After endothelial cellular networks appeared, trophoblast derived HTR‐8/SVneo cells were co‐cultured in the presence of clinically relevant doses of methyldopa, labetalol, hydralazine or clonidine for 24 hours. Cells were retrieved from the Matrigel to extract mRNA and eNOS and iNOS expression were examined by quantitative PCR. Methyldopa, labetalol, hydralazine and clonidine reversed the inhibitory effect of TNF‐α on eNOS mRNA expression. After pre‐incubating endothelial cells with TNF‐α and sFlt‐1, all the medications except methyldopa lost their effect on eNOS mRNA expression. In the absence of TNF‐α, antihypertensive medications did not change eNOS expression. The mRNA expression of iNOS was not affected by TNF‐α or any medications. This study shows that selected antihypertensive medications used in the treatment of hypertension in pregnancy increase eNOS expression in vitro when induced by the inflammatory TNF‐α. The anti‐angiogenic molecule sFlt‐1 may antagonise the potential benefit of these medications by interfering with the NOS pathway.
    February 14, 2017   doi: 10.1111/1440-1681.12712   open full text
  • Hypoxia‐inducible factors as neuroprotective agent in Alzheimer's disease.
    Ben Sundra Ashok, Thekkuttuparambil Ananthanarayanan Ajith, Senthilkumar Sivanesan.
    Clinical and Experimental Pharmacology and Physiology. February 14, 2017
    Beta amyloid (Aβ)‐42 peptide and phosphorylated tau protein have been demonstrated as the pathological hallmarks of Alzheimer's disease (AD). A gradual decline of oxygen and glucose supply to the brain during aging or hypoxia was manifested as a contributing factor to hypometabolism. The brain regions susceptible to hypometabolism are the hippocampus, entorhinal cortex and cognition‐associated neocortical regions like parietal, temporal and frontal cortex. In AD patients, the brain regions with hypometabolism can trigger overexpression of amyloid precursor protein and decrease the clearance of Aβ. Aβ and hypoxia can evoke inflammation, oxidative stress and finally neuronal cell death. Among the transcription factors involved in the compensatory mechanism, hypoxia‐inducible factor‐1 alpha (HIF‐1α) has a major role in the cellular adaptation by inducing the expression of several proteins, including vascular endothelial growth factor, erythropoietin and inducible nitric oxide synthase. Therefore, maintaining the HIF‐1α level by inhibiting the prolyl 4‐hydroxylase was effective to attenuate the nerve damage during hypoxia and postpone the incidence of AD. Agents such as iron chelators, and heavy metals like cobalt and nickel were demonstrated to be effective in maintaining the HIF‐1α level in the nerve. This review article discusses the possible role of HIF‐1α as a neuroprotector in AD and the future perspectives.
    February 14, 2017   doi: 10.1111/1440-1681.12717   open full text
  • Aliskiren protects against myocardial ischaemia‐reperfusion injury via an endothelial nitric oxide synthase dependent manner.
    Yu Chen, Guoliang Meng, Wenli Bai, Yan Ma, Liping Xie, Naila Altaf, Yanning Qian, Yi Han, Yong Ji.
    Clinical and Experimental Pharmacology and Physiology. January 24, 2017
    Aliskiren, a direct renin inhibitor, has shown potent ability to attenuate hypertension. Our previous research has found that aliskiren protected against myocardial ischaemia‐reperfusion (I/R) injury and enhanced phosphorylation of endothelial nitric oxide synthase (eNOS) in spontaneously hypertensive rats. However, whether the cardioprotective effect of aliskiren against myocardial I/R injury was eNOS‐dependent is unknown. In the present study, 12‐week‐old male eNOS knockout (eNOS−/−) and wild‐type C57BL/6J mice (WT) were orally administrated with the dose of 50 mg/kg per day of aliskiren. After a 4‐week treatment, aliskiren decreased blood pressure in eNOS−/− mice, and reduced renin‐angiotension II levels in both eNOS−/− and WT mice. Aliskiren also improved left ventricular ejection fraction (EF) and fractional shortening (FS), decreased myocardial infarct size, reduced creatine kinase (CK) and lactate dehydrogenase (LDH) activity in plasma, attenuated dihydroethidium (DHE) fluorescence and levels of malondialdehyde (MDA), enhanced superoxide dismutase (SOD) activity and total antioxidant capacity (T‐AOC) in myocardium, increased SOD and thioredoxin (Trx) proteins expression in WT mice subjected to 30 minutes of ischaemia followed by reperfusion for 24 hours. However, aliskiren failed to restore all of the above indices in eNOS−/− mice subjected to the same I/R injury. Our study indicated that aliskiren protected against myocardial I/R injury via an eNOS dependent manner.
    January 24, 2017   doi: 10.1111/1440-1681.12692   open full text
  • Renal function in HIV/HBV co‐infected and HBV mono‐infected patients on a long‐term treatment with tenofovir in real life setting.
    Laura Milazzo, Cristina Gervasoni, Felicia Stefania Falvella, Dario Cattaneo, Cristina Mazzali, Paola Ronzi, Francesca Binda, Stefania Cheli, Salvatore Sollima, Spinello Antinori.
    Clinical and Experimental Pharmacology and Physiology. January 24, 2017
    The human immunodeficiency virus (HIV)/hepatitis B virus (HBV) co‐infection is likely to be associated with an increased risk of kidney disease, due to the additional factors that may affect renal function in the HIV population. We aimed to evaluate renal toxicity in HIV/HBV and HBV mono‐infected patients on long‐term therapy with tenofovir (TDF) and to explore the association of polymorphisms in ATP‐binding cassette (ABCC)2, ABCC4, ABCC10 with the development of renal dysfunction. From September 2006 to November 2014, 44 HIV/HBV co‐infected and 34 HBV mono‐infected patients were commenced on TDF. Data of renal safety were retrospectively collected and analyzed. ABCC2, ABCC4 and ABCC10 genotypes were identified by real‐time PCR. Over 60 months of observation, there was a significant increase in mean creatinine levels from baseline (P<.01) that was not significantly different between the two study groups. Moreover, a significant decline in estimated glomerular filtration rate (eGFR) was observed from baseline (P<.01), and it was significantly greater in HBV mono‐infected than co‐infected patients (P=.03). The distribution of ABCC2, ABCC4 and ABCC10 genotypes among a subgroup of 34 patients did not show significant association with eGFR decline <90 mL/min per 1.73 m2. Although our findings showed a statistically significant decrease in eGFR with long‐term use of TDF, its clinical impact seems to be modest. The role of genetic factors to identify patients at greater risk for developing tenofovir‐induced renal toxicity needs to be further investigated.
    January 24, 2017   doi: 10.1111/1440-1681.12691   open full text
  • Drug resistance and cancer stem cells: the shared but distinct roles of hypoxia‐inducible factors HIF1α and HIF2α.
    Jennifer Petra Schöning, Michael Monteiro, Wenyi Gu.
    Clinical and Experimental Pharmacology and Physiology. January 24, 2017
    Chemotherapy resistance is a major contributor to poor treatment responses and tumour relapse, the development of which has been strongly linked to the action of cancer stem cells (CSCs). Mounting evidence suggests that CSCs are reliant on low oxygen conditions and hypoxia‐inducible factors 1α and 2α (HIF1α and HIF2α) to maintain their stem cell features. Research in the last decade has begun to clarify the functional differences between the two HIFα subtypes (HIFαs). Here, we review and discuss these differences in relation to CSC‐associated drug resistance. Both HIFαs contribute to CSC survival but play different roles –HIF1α being more responsible for survival functions and HIF2α for stemness traits such as self‐renewal – and are sensitive to different degrees of hypoxia. Failure to account for physiologically relevant oxygen concentrations in many studies may influence the current understanding of the roles of HIFαs. We also discuss how hypoxia and HIFαs contribute to CSC drug resistance via promotion of ABC drug transporters Breast cancer resistance protein (BCRP), MDR1, and MRP1 and through maintenance of quiescence. Additionally, we explore the PI3K/AKT cell survival pathway that may support refractory cancer by promoting CSCs and activating both HIF1α and HIF2α. Accordingly, HIF1α and HIF2α inhibition, potentially via PI3K/AKT inhibitors, could reduce chemotherapy resistance and prevent cancer relapse.
    January 24, 2017   doi: 10.1111/1440-1681.12693   open full text
  • Effects of high‐sodium intake on systemic blood pressure and vascular responses in spontaneously diabetic WBN/Kob‐Leprfa/fa rats.
    Yoshiichi Takagi, Haruno Kadowaki, Ikumi Kobayashi, Kaoru Ito, Katsuaki Ito, Mitsuyuki Shirai, Fumitoshi Asai.
    Clinical and Experimental Pharmacology and Physiology. January 24, 2017
    The prevalence of type 2 diabetes mellitus (T2DM) and hypertension has markedly increased worldwide. The purpose of the present study was to examine the effects of a high‐salt intake on the systolic blood pressure (SBP) and vascular responses in WBN/Kob‐Leprfa/fa (WBKDF) rats, a new spontaneous animal model of T2DM. Male WBKDF rats and age‐matched Wistar rats at 6 weeks of age were each divided into two groups and fed either a normal‐sodium (NS, 0.26%) diet or high‐sodium (HS, 8%) diet for 14 weeks: (i) Wistar rats on NS diet (Wistar‐NS); (ii) Wistar rats on HS diet (Wistar‐HS); (iii) WBKDF rats on NS diet (WBKDF‐NS); (iv) WBKDF rats on HS diets (WBKDF‐HS). Neither WBKDF‐NS nor Wistar‐NS rats showed significant changes in SBP throughout the experiment, but both WBKDF‐HS and Wistar‐HS exhibited significant elevation of SBP, which was more prominent (P<.01) in WBKDF‐HS than in Wistar‐HS. Phenylephrine‐induced contractions of isolated thoracic aortic rings were significantly (P<.01) enhanced in WBKDF‐HS and Wistar‐HS compared with the respective strain of rats on the NS diet. In contrast, acetylcholine‐ and nitroprusside‐induced relaxation were significantly (P<.01) diminished in both WBKDF‐HS and Wistar‐HS, and these HS diet‐induced changes were more profound (P<.01) in WBKDF rats than in Wistar rats. Significantly (P<.05) higher plasma concentrations of 8‐iso‐prostaglandin F2α and sodium ions were observed in WBKDF‐HS than in Wistar‐HS. The current study demonstrated that WBKDF‐HS rats developed salt‐sensitive hypertension associated with vascular dysfunction. The WBKDF rat may be a useful model for investigating the etiology of hypertension with T2DM.
    January 24, 2017   doi: 10.1111/1440-1681.12700   open full text
  • Connexin32 deficiency is associated with liver injury, inflammation and oxidative stress in experimental non‐alcoholic steatohepatitis.
    Taynã Cristina Tiburcio, Joost Willebrords, Tereza Cristina Silva, Isabel Veloso Alves Pereira, Marina Sayuri Nogueira, Sara Crespo Yanguas, Michaël Maes, Elisangela dos Anjos Silva, Maria Lucia Zaidan Dagli, Inar Alves Castro, Cláudia Pinto Oliveira, Mathieu Vinken, Bruno Cogliati.
    Clinical and Experimental Pharmacology and Physiology. January 24, 2017
    Non‐alcoholic steatohepatitis is a highly prevalent liver pathology featured by hepatocellular fat deposition and inflammation. Connexin32, which is the major building block of hepatocellular gap junctions, has a protective role in hepatocarcinogenesis and is downregulated in chronic liver diseases. However, the role of connexin32 in non‐alcoholic steatohepatitis remains unclear. Connexin32−/− mice and their wild‐type littermates were fed a choline‐deficient high‐fat diet. The manifestation of non‐alcoholic steatohepatitis was evaluated based on a battery of clinically relevant read‐outs, including histopathological examination, diverse indicators of inflammation and liver damage, in‐depth lipid analysis, assessment of oxidative stress, insulin and glucose tolerance, liver regeneration and lipid‐related biomarkers. Overall, more pronounced liver damage, inflammation and oxidative stress were observed in connexin32−/− mice compared to wild‐type animals. No differences were found in insulin and glucose tolerance measurements and liver regeneration. However, two lipid‐related genes, srebf1 and fabp3, were upregulated in Cx32−/− mice in comparison with wild‐type animals. These findings suggest that connexin32‐based signalling is not directly involved in steatosis as such, but rather in the sequelae of this process, which underlie progression of non‐alcoholic steatohepatitis.
    January 24, 2017   doi: 10.1111/1440-1681.12701   open full text
  • Maternal Na+ intake induces renal function injury in rats prevented by a short‐term angiotensin converting enzyme inhibitor.
    Natalie E Ribeiro, Edjair V Cabral, Regina S Aires, Leucio D Vieira‐Filho, Valdilene S Ribeiro, Daianna RM Gonçalves, Luis PNC Borges, Ismaela MF Melo, Cintia GM Ferreira, Valeria Wanderley‐Teixeira, Álvaro AC Teixeira, Anísio F Soares, Ana D Paixão.
    Clinical and Experimental Pharmacology and Physiology. January 24, 2017
    The Na+‐ATPase, a secondary pump in the proximal tubule, is only weakly responsive to angiotensin II in adult offspring exposed perinatally to high Na+ intake. We have investigated whether the offspring from mothers given 0.3 mol/L NaCl show an ineffective angiotensin II action to increase in blood pressure. It was hypothesized that functional alterations at adult life are associated with the number of angiotensin II‐positive cells in the developing kidney, with increased oxidative stress in maternal/foetal organs, or with morphometrical changes in placentas. Wistar female rats were maintained on 0.3 mol/L NaCl in their drinking water from 20 days before conception until weaning. After weaning, some of the male offspring were treated with enalapril for 21 days. Glomerular filtration rate was recorded up to 210 days of age, when mean arterial pressure was measured after infusion of angiotensin II. To investigate the placenta and foetal kidneys, mothers on tap water or NaCl were also treated with alpha‐tocopherol, pregnancy being interrupted on the 20th day. There were no changes in the number of cells positive for angiotensin II in the foetal kidney and unchanged lipid peroxidation in the placenta of offspring exposed to NaCl, but the intermediate trophoblast area in the junctional zone was increased, possibly reducing maternal–foetal exchange. Glomerular filtration rate was reduced and there was an attenuated effect of angiotensin II on elevation of blood pressure, which could be mediated by an elevated angiotensin II during early life, once these disturbances had been prevented by early and short‐term treatment with enalapril.
    January 24, 2017   doi: 10.1111/1440-1681.12702   open full text
  • Reduction of the foreign body response and neuroprotection by apyrase and minocycline in chronic cannula implantation in the rat brain.
    Linda Hayn, Linda Deppermann, Michael Koch.
    Clinical and Experimental Pharmacology and Physiology. January 24, 2017
    Implantation of electrodes or cannulae into the brain is accompanied by a tissue response referred to as foreign body response. Adenosine triphosphate (ATP) is one of the signalling molecules released by injured cells which mediate the chemoattraction of microglial cells. The constitutive release of pro‐inflammatory and cytotoxic substances by microglial cells in chronic implants exacerbates neuronal cell death and the immune response. This study aimed to interfere with the initial events of the foreign body response in order to mitigate neurotoxicity and inflammation. For this purpose, the ATP‐hydrolysing enzyme apyrase and the antibiotic minocycline with a broad range of anti‐inflammatory, anti‐apoptotic and glutamate‐antagonist properties were locally infused during cannula implantation in the caudal forelimb area of the motor cortex in Lister Hooded rats. The rats’ motor performance was assessed in a skilled reaching task and the distribution of neurons and glial cells in the vicinity of the implant was examined 2 and 6 weeks post‐implantation. Apyrase as well as minocycline increased the number of surviving neurons and reduced microglial activation. Moreover, minocycline improved the motor performance and, additionally, caused a temporary reduction in astrogliosis, suggesting it as a possible therapeutic candidate to improve the biocompatibility of chronic brain implants.
    January 24, 2017   doi: 10.1111/1440-1681.12703   open full text
  • GRK2/β‐arrestin mediates arginine vasopressin‐induced cardiac fibroblast proliferation.
    Yunxuan Chen, Feifei Xu, Lingling Zhang, Xiaojun Wang, Yifan Wang, Anthony Yiu‐Ho Woo, Weizhong Zhu.
    Clinical and Experimental Pharmacology and Physiology. January 24, 2017
    Cardiac fibrosis is a pathological feature commonly found in hearts exposed to haemodynamic orneurohormonal stress. Elevated levels of arginine vasopressin (AVP) are closely associated with the progression of heart failure and could be an underlying cause of cardiac fibrosis. The aim of this study is to characterize the effect of AVP on neonatal rat cardiac fibroblasts (NRCFs) and to illustrate its signalling mechanism. The proliferative effect of AVP was assessed by methylthiazolyldiphenyl‐tetrazolium assay and 5‐bromo‐2′‐deoxyuridine (BrdU) incorporation assay, and the amounts of cellular signalling proteins α‐smooth muscle actin (α‐SMA), matrix metalloproteinase (MMP) 2, MMP9, and phosphorylated ERK1/2 were determined by western blotting. AVP, in a time‐ and concentration‐dependent manner, promoted NRCF proliferation and the expression of MMP2 and MMP9. Inhibition of G protein‐coupled receptor kinase2 (GRK2) by the inhibitory peptide GRK2‐Ct or knock‐down of GRK2 suppressed AVP‐induced BrdU incorporation and the expression of MMP2 and α‐SMA in NRCFs. Moreover, shRNA‐mediated silencing of β‐arrestin1 or β‐arrestin 2 abolished AVP‐induced BrdU incorporation and MMP2 expression. AVP‐induced NRCF proliferation depended on the phosphorylation of ERK1/2, and inhibition of GRK2 or silencing of β‐arrestins blocked AVP‐induced ERK1/2 phosphorylation. The effects of AVP on NRCF proliferation and α‐SMA expression were blocked by SR45059, a vasopressin receptor type1A (V1AR) selective antagonist. In conclusion, AVP promotes NRCF proliferation through V1AR‐mediated GRK2/β‐arrestin/ERK1/2 signalling.
    January 24, 2017   doi: 10.1111/1440-1681.12696   open full text
  • The sensitivity of head and neck carcinoma cells to statins is related to the expression of their Ras expression status, and statin‐induced apoptosis is mediated via suppression of the Ras/ERK and Ras/mTOR pathways.
    Masanobu Tsubaki, Daichiro Fujiwara, Tomoya Takeda, Toshiki Kino, Yoshika Tomonari, Tatsuki Itoh, Motohiro Imano, Takao Satou, Katsuhiko Sakaguchi, Shozo Nishida.
    Clinical and Experimental Pharmacology and Physiology. January 24, 2017
    Statins induce apoptosis of tumour cells by inhibiting the prenylation of small G‐proteins. However, the details of the apoptosis‐inducing mechanisms remain poorly understood. The present study showed that the induction of apoptosis by statins in four different human head and neck squamous cell carcinoma (HNSCC) cell lines, HSC‐3, HEp‐2, Ca9‐22, and SAS cells was mediated by increased caspase‐3 activity. Statins induced apoptosis by the suppression of geranylgeranyl pyrophosphate biosynthesis. Furthermore, statins decreased the levels of phosphorylated ERK and mTOR by inhibiting the membrane localization of Ras and enhancing Bim expression in HSC‐3 and HEp‐2 cells. We also found that in all the cell types analyzed, the IC50 values for fluvastatin and simvastatin were highest in HEp‐2 cells. In addition, HSC‐3, Ca9‐22, and SAS cells had higher Ras expression and membrane localization, higher activation of ERK1/2 and mTOR, and lower levels of Bim expression than HEp‐2 cells. Our results indicate that statins induce apoptosis by increasing the activation of caspase‐3 and by enhancing Bim expression through inhibition of the Ras/ERK and Ras/mTOR pathways. Furthermore, the sensitivity of HNSCC cells to statin treatment was closely related to Ras expression and prenylation levels, indicating that statins may act more effectively against tumours with high Ras expression and Ras‐variability. Therefore, our findings support the use of statins as potential anticancer agents.
    January 24, 2017   doi: 10.1111/1440-1681.12690   open full text
  • Decreased percentage of NKG2D+NK cells in patients with incident onset of Type 1 Diabetes.
    Yupan Zhang, Haifeng Wang, Xiaoqian Lou, Xiaozhang Qu, Lichao Gao, Xiaolei Liu, Man Li, Hui Guo, Yanfang Jiang.
    Clinical and Experimental Pharmacology and Physiology. January 24, 2017
    Type 1 diabetes mellitus (T1DM) is characterized by absolute insulin deficiency owing to autoimmune destruction of the pancreatic β cells. A significant decrease in natural killer (NK) cells in peripheral blood has been observed in patients with untreated T1DM. In the present study, we aimed to explore the role of NK cells and their subsets in young T1DM patients. A total of 30 children and adolescents with untreated T1DM and 27 healthy controls (HC) were recruited in this study. Flow cytometry analysis indicated that the percentage of peripheral blood CD3‐CD56+ NK cells and NK cells subsets (CD56bright, CD56dim and CD56neg), were significantly decreased in the T1DM patients compared to healthy controls. In addition, the percentage of inducible CD107a+ and IFN‐γ‐secreting NK cells was significantly decreased compared to HC. Interestingly, the percentage of NKG2D+ NK cells negatively correlated with the level of serum TCHOL and TG in T1DM patients. Our data indicate that decreased number and impaired function of NK cells may have a role in the pathogenesis of T1DM.
    January 24, 2017   doi: 10.1111/1440-1681.12699   open full text
  • β‐eudesmol suppresses allergic reactions via inhibiting mast cell degranulation.
    Na‐Ra Han, Phil‐Dong Moon, Ka‐Jung Ryu, Jae‐Bum Jang, Hyung‐Min Kim, Hyun‐Ja Jeong.
    Clinical and Experimental Pharmacology and Physiology. January 24, 2017
    The regulatory effect of β‐eudesmol, which is an active constituent of Pyeongwee‐San (KMP6), is evaluated for allergic reactions induced by mast cell degranulation. Phorbol 12‐myristate 13‐acetate (PMA) plus calcium ionophore A23187‐stimulated human mast cell line, HMC‐1 cells, and compound 48/80‐stimulated rat peritoneal mast cells (RPMCs) are used as the in vitro models; mice models of systemic anaphylaxis, ear swelling, and IgE‐dependent passive cutaneous anaphylaxis (PCA) are used as the in vivo allergic models. The results demonstrate that β‐eudesmol suppressed the histamine and tryptase releases from the PMA plus calcium ionophore A23187‐stimulated HMC‐1 cells. β‐eudesmol inhibits the expression and activity of histidine decarboxylase in the activated HMC‐1 cells. In addition, β‐eudesmol inhibits the levels of histamine and tryptase released from the compound 48/80‐stimulated RPMCs. Furthermore, β‐eudesmol decreases the intracellular calcium level in the activated RPMCs. β‐eudesmol also decreases the compound 48/80‐induced mortality and ear swelling response. β‐eudesmol suppresses the serum levels of histamine, IgE, interleukin (IL)‐1β, IL‐4, IL‐5, IL‐6, IL‐13, and vascular endothelial growth factor (VEGF) under PCA mice as well as PCA reactions. Therefore, the results from this study indicate the potential of β‐eudesmol as an anti‐allergic drug with respect to its pharmacological properties against mast cell‐mediated allergic reactions.
    January 24, 2017   doi: 10.1111/1440-1681.12698   open full text
  • Impulse control disorder, lysosomal malfunction and ATP13A2 insufficiency in Parkinsonism.
    Jun‐Ping Liu, Jianfeng Li, Yanhua Lu, Lihui Wang, Gang Chen.
    Clinical and Experimental Pharmacology and Physiology. January 24, 2017
    Lysosomal transport of cargos in neurons is essential for neuronal proteostasis, transmission and functional motors and behaviours. Lysosomal malfunction including storage disorders is involved in the pathogenesis of Parkinson's disease (PD). Given the unclear molecular mechanisms of diverse defects in PD phenotypes, especially behavioural deficits, this mini review explores the cellular contexts of PD impulse control disorders and the molecular aspects of lysosomal cross‐membrane transports. Focuses are paid to trace metal involvements in α‐synuclein assembly in Lewy bodies, the functions and molecular interactions of ATP13A2 as ATPase transporters in lysosomal membranes for cross‐membrane trafficking and lysosomal homeostasis, and our current understandings of the neural circuits in ICD. Erroneously polarized distributions of cargos such as metals and lipids on each side of lysosomal membranes triggered by gene mutations and deregulated expression of ATP13A2 may thus instigate sensing protein structural changes such as aggregations, organelle degeneration, and specific neuronal ageing and death in Parkinsonism.
    January 24, 2017   doi: 10.1111/1440-1681.12714   open full text
  • Naltrexone changes the expression of lipid metabolism‐related proteins in the endoplasmic reticulum stress induced hepatic steatosis in mice.
    Azam Moslehi, Fatemeh Nabavizadeh, Ali Zekri, Fatemeh Amiri.
    Clinical and Experimental Pharmacology and Physiology. January 24, 2017
    Endoplasmic reticulum (ER) stress is closely associated with several chronic diseases such as obesity, atherosclerosis, type 2 diabetes, and hepatic steatosis. Steatosis in hepatocytes may also lead to disorders such as nonalcoholic fatty liver disease (NAFLD) and nonalcoholic steatohepatitis (NASH), fibrosis, and possibly cirrhosis. Opioid peptides are involved in triglyceride and cholesterol dysregulation. Naltrexone also attenuates ER stress induced hepatic steatosis in mice. In this study, we evaluated the effects of naltrexone on the expression of lipid metabolism‐related nuclear factors and enzymes in the ER stress induced hepatic steatosis. C57/BL6 mice received saline, DMSO and naltrexone as control groups. In a fourth group, ER stress was induced by tunicamycin (TM) injection and in the last group, naltrexone was given before TM administration. Histopathological evaluations, real‐time RT‐PCR and western blot were performed. We found that GRP78, IRE1α, PERK and ATF6 gene expression and steatosis significantly reduced in naltrexone treated animals. Naltrexone alleviated the gene and protein expression of SREBP1c. Expression of ACAT1, apolipoprotein B (ApoB) and PPARα also increased after naltrexone treatment. In conclusion, this study, for the first time, shows that naltrexone has a considerable role in attenuation of ER stress‐induced liver injury.
    January 24, 2017   doi: 10.1111/1440-1681.12695   open full text
  • A tumour‐promoting role of Th9 cells in hepatocellular carcinoma through CCL20 and STAT3 pathways.
    Hongwu Tan, Shuyun Wang, Ludong Zhao.
    Clinical and Experimental Pharmacology and Physiology. January 24, 2017
    Hepatocellular carcinoma (HCC) is a common and aggressive human malignancy. An imperative demand is present for a better understanding of the functions and regulations of the immune system and how they affect HCC pathogenesis. Recently, a group of interleukin 9 (IL‐9)‐producing CD4+ T cells, termed Th9 cells, has been described in mice and humans with both tumour‐inhibiting as well as tumour‐promoting effects. The specific roles of Th9 cells in human HCC are not entirely understood. Here, we examined the frequencies and functions of IL‐9‐producing Th9 cells in HCC patients. We found that the frequencies of circulating IL‐9‐producing Th9 cells were significantly higher in HCC patients compared to in healthy individuals. In HCC patients, the frequencies of IL‐9‐producing Th9 cells were significantly higher in peritumour and tumour tissues than in unaffected liver tissues. Interestingly, HCC patients with higher tumour‐infiltrating Th9 frequency had significantly shorter disease‐free survival period after resection. Previously, high expression of CCL20 was associated with poor prognosis in HCC. CCL20 also induced epithelial‐mesenchymal transition‐like changes in HCC cells. We found that incubation of primary HCC cells with autologous Th9 significantly elevated the CCL20 production from tumour cells, which could be partially inhibited by suppressing STAT3. Together, this study suggested a tumour‐promoting role of Th9 cells in HCC.
    January 24, 2017   doi: 10.1111/1440-1681.12689   open full text
  • Mycophenolate mofetil improves renal haemodynamics, microvascular oxygenation, and inflammation in a rat model of supra‐renal aortic clamping‐mediated renal ischaemia reperfusion injury.
    Bulent Ergin, Michal Heger, Asli Kandil, Cihan Demirci‐Tansel, Can Ince.
    Clinical and Experimental Pharmacology and Physiology. January 24, 2017
    Ischaemia/reperfusion (I/R) is one of the main causes of acute kidney injury (AKI), which is characterized by sterile inflammation and oxidative stress. Immune cell activation can provoke overproduction of inflammatory mediators and reactive oxygen species (ROS), leading to perturbation of the microcirculation and tissue oxygenation associated with local and remote tissue injury. This study investigated whether the clinically employed immunosuppressant mycophenolate mofetil (MMF) was able to reduce I/R‐induced renal oxygenation defects and oxidative stress by preventing sterile inflammation. Rats were divided into three groups (n=6/group): (1) a sham‐operated control group; (2) a group subjected to renal I/R alone (I/R); and (3) a group subjected to I/R and MMF treatment (20 mg/kg prior to I/R) (I/R+MMF). Ischaemia was induced by a vascular occluder placed on the abdominal aorta for 30 minutes, followed by 120 minutes of reperfusion. Renal I/R deteriorated renal oxygenation (P<.001) and oxygen delivery (P<.01), reduced creatinine clearance (P<.01) and tubular sodium reabsorption (P<.001), and increased iNOS, renal tissue injury markers (P<.001), and IL‐6 (P<.001). Oral MMF administration prior to insult restored renal cortical oxygenation (P<.05) and iNOS, renal injury markers, and inflammation parameters (P<.001) to near‐baseline levels without affecting renal function. MMF exerted a prophylactic effect on renal microvascular oxygenation and abrogated tissue inflammation and renal injury following lower body I/R‐induced AKI. These findings may have clinical implications during major vascular or renal transplant surgery.
    January 24, 2017   doi: 10.1111/1440-1681.12687   open full text
  • Gremlin1 promotes carcinogenesis of glioma in vitro.
    Yongchang Guan, Wen Cheng, Cunyi Zou, Tingzhong Wang, Zhi Cao.
    Clinical and Experimental Pharmacology and Physiology. January 24, 2017
    As the most prevalent and lethal type of brain tumours, gliomas, especially malignant ones, are relatively resistant to conventional therapies. Gremlin 1 (GREM1) is a secreted glycoprotein that is implicated in the maintenance of cancer stem cells in tumour hierarchy. In the current study, the role of GREM1 in the carcinogenesis of glioma was studied using a knockdown approach. We first examined the expression level of GREM1 in the clinical samples, and then evaluated the effect of GREM1 knockdown on the viability and colony formation capacity of U87‐MG cells. Moreover, the migration ability, invasiveness, cell cycle, and apoptosis of GREM1‐silenced cells were assessed. Furthermore, the involvement of functional GREM1 in the epithelial‐mesenchymal transition (EMT) process of glioma was investigated by detecting the expression levels of glioma‐associated oncogene homologue 3 (GLI3) and EMT‐related molecules. Our results demonstrated that knockdown of GREM1 reduced cell viability, suppressed migration and invasion, and inhibited GLI3 expression and the EMT process in U87‐MG cells. Meanwhile, GREM1 silencing promoted apoptosis in U87‐MG cells through the accumulation of Bax, cleaved caspase‐3, and cleaved poly (ADP‐ribose) polymerase (PARP) as well as the downregulation of Bcl‐2. In addition, GREM1 knockdown abolished transforming growth factor (TGF)‐β1‐mediated activation of the Smad pathway, which may underlie the mechanism of GREM1‐regulated EMT. In conclusion, GREM1 plays an important role in the development of glioma, and it may serve as a potential target in glioma therapy.
    January 24, 2017   doi: 10.1111/1440-1681.12697   open full text
  • Zinc oxide nanoparticles as a novel anticancer approach; in vitro and in vivo evidence.
    Hesham Fathy Hassan Hassan, Ahmed Mohamed Mansour, Amira Morad Hussein Abo‐Youssef, Bakheet E M Elsadek, Basim Anwar Shehata Messiha.
    Clinical and Experimental Pharmacology and Physiology. January 24, 2017
    Currently, the outcomes of conventional chemotherapeutic approaches are unsatisfactory. Clinical application of nanoparticles seems promising. We aim to evaluate the possible antitumor activity of zinc oxide nanoparticles (ZnONPs) as a chemotherapeutic approach in in vitro and in vivo experimental models. An in vitro study was performed on three different cell lines, namely human hepatocellular carcinoma (HEPG2), human prostate cancer (PC3), and none‐small cell lung cancer (A549) cell lines. An in vivo study using diethylnitrosamine (DENA)‐induced HCC in adult male Wistar rats was conducted to investigate the potential antitumor activity of ZnONPs in HCC and the possible underlying mechanisms. Hepatocellular carcinoma (HCC) was induced by oral administration of DENA given in drinking water (100 mg/L) for 8 weeks. Rats were allocated into four groups, namely a control group, an HCC control group receiving DENA alone, a ZnONPs (10 μg/kg per week, intravenous (i.v.) for 1 month) control group, and a ZnONPs treatment group (receiving ZnONPs + DENA). ZnONPs significantly reduced the elevated serum levels of HCC‐related tumor markers alphafetoprotein and alpha‐l‐fucosidase and the apoptotic marker caspase‐3 compared with the untreated HCC rats. In addition, treatment with ZnONPs significantly decreased the elevated levels of hepatocyte integrity and oxidative stress markers as compared with the untreated HCC control group. Furthermore, the histopathological study revealed anaplasia and fibrous degenerations which were significantly corrected by ZnONPs treatment. In conclusion, administration of ZnONPs exhibited a promising preclinical anticancer efficacy in HCC and could be considered as a novel strategy for the treatment HCC in clinical practices.
    January 24, 2017   doi: 10.1111/1440-1681.12681   open full text
  • R1 autonomic nervous system in acute kidney injury.
    Dagmara Hering, Pawel J Winklewski.
    Clinical and Experimental Pharmacology and Physiology. January 24, 2017
    Acute kidney injury (AKI) is a rapid loss of kidney function resulting in accumulation of end metabolic products and associated abnormalities in fluid, electrolyte and acid‐base homeostasis. The pathophysiology of AKI is complex and multifactorial involving numerous vascular, tubular and inflammatory pathways. Neurohumoral activation with heightened activity of the sympathetic nervous system and renin‐angiotensin‐aldosterone system play a critical role in this scenario. Inflammation and/or local renal ischaemia are underlying mechanisms triggering renal tissue hypoxia and resultant renal microcirculation dysfunction; a common feature of AKI occurring in numerous clinical conditions leading to a high morbidity and mortality rate. The contribution of renal nerves to the pathogenesis of AKI has been extensively demonstrated in a series of experimental models over the past decades. While this has led to better knowledge of the pathogenesis of human AKI, therapeutic approaches to improve patient outcomes are scarce. Restoration of autonomic regulatory function with vagal nerve stimulation resulting in anti‐inflammatory effects and modulation of centrally‐mediated mechanisms could be of clinical relevance. Evidence from experimental studies suggests that a therapeutic splenic ultrasound approach may prevent AKI via activation of the cholinergic anti‐inflammatory pathway. This review briefly summarizes renal nerve anatomy, basic insights into neural control of renal function in the physiological state and the involvement of the autonomic nervous system in the pathophysiology of AKI chiefly due to sepsis, cardiopulmonary bypass and ischaemia/reperfusion experimental model. Finally, potentially preventive experimental pre‐clinical approaches for the treatment of AKI aimed at sympathetic inhibition and/or parasympathetic stimulation are presented.
    January 24, 2017   doi: 10.1111/1440-1681.12694   open full text
  • Enhanced synergistic anti‐Lewis lung carcinoma effect of a DNA vaccine harboring a MUC1‐VEGFR2 fusion gene used with GM‐CSF as an adjuvant.
    Junzhong Ruan, Yong Duan, Fugen Li, Zitong Wang.
    Clinical and Experimental Pharmacology and Physiology. December 27, 2016
    In order to achieve a synergistic effect on anti‐tumour and anti‐angiogenesis activity, we designed and constructed a DNA vaccine that expresses MUC1and VEGFR2 in the same reading frame. The aim of this study was to investigate the anti‐tumour activity of this DNA vaccine. Furthermore, we also investigated the enhanced synergistic anti‐Lewis lung carcinoma effect of this DNA vaccine by using GM‐CSF as an adjuvant. A series of DNA plasmids encoding MUC1, VEGFR2, GM‐CSF, and their conjugates were constructed and injected into mice intramuscularly (i.m.) followed by an electric pulse. The humoral and cellular immune responses after immunization were detected by enzyme‐linked immunosorbent assay (ELISA) and enzyme‐linked immunospot (ELISPOT), respectively. To evaluate the anti‐tumour efficacy of these plasmids, murine models with MUC1‐expressing tumours were generated. After injection into the tumour‐bearing mouse model, the plasmid carrying the fusion gene of MUC1 and VEGFR2 showed stronger inhibition of tumour growth than the plasmid expressing MUC1 or VEGFR2 alone, which indicated that MUC1 and VEGFR2 could exert a synergistic anti‐tumour effect. Furthermore, mice vaccinated with the combination of the GM‐CSF expressing plasmid and the plasmid carrying the fusion gene of MUC1 and VEGFR2 showed an increased inhibition in the growth of MUC1‐expressing tumours and prolonged mouse survival. These observations emphasize the potential of the synergistic anti‐tumour and anti‐angiogenesis strategy used in DNA vaccines, and the potential of the GM‐CSF gene as an adjuvant for DNA vaccines, which could represent a promising approach for tumour immunotherapy.
    December 27, 2016   doi: 10.1111/1440-1681.12654   open full text
  • Pulmonary vascular inflammation: effect of TLR signalling on angiopoietin/TIE regulation.
    Tobias Hilbert, Kathrin Dornbusch, Georg Baumgarten, Andreas Hoeft, Stilla Frede, Sven Klaschik.
    Clinical and Experimental Pharmacology and Physiology. December 27, 2016
    Increased pulmonary vascular resistance is a critical complication in sepsis. Toll‐like receptor (TLR) as well as angiopoietin (ANG) signalling both contribute to the emergence of pulmonary arterial hypertension. We hypothesized that TLR stimulation by bacterial ligands directly affects expression and secretion of ligands and receptors of the angiopoietin/TIE axis. Microvascular endothelial (HPMEC) and smooth muscle cells (SMC) of pulmonary origin were incubated with thrombin and with ligands for TLR2, ‐4, ‐5, and ‐9. Expression and secretion of ANG1, ‐2, TIE2 and IL‐8 were determined using quantitative real‐time PCR and ELISA. TLR stimulation had no impact either on the expression of ANG2 and TIE2 in HPMEC or on that of ANG1 in SMC. However, overall levels of both released ANG1 and ‐2 were halved upon stimulation with the TLR9 ligand CpG, and ANG2 release was significantly enhanced by TLR4 activation when initially provoked by sequentially performed stimulation. Furthermore, enhanced ANG2 activity increased endothelial permeability, as demonstrated in an in vitro transwell assay. We conclude that sole TLR stimulation by bacterial ligands plays no significant role for altered expression and secretion of ANG1, ‐2 and TIE2 in human pulmonary vascular cells. The interplay between various stimuli is required to induce imbalances between ANG1 and ‐2.
    December 27, 2016   doi: 10.1111/1440-1681.12680   open full text
  • Response surface modelling of the pharmacodynamic interaction between propofol and remifentanil in patients undergoing anaesthesia.
    SangMin Choe, Byung‐Moon Choi, Yong‐Hun Lee, Soo‐Han Lee, Eun‐Kyung Lee, Ki‐Seong Kim, Gyu‐Jeong Noh.
    Clinical and Experimental Pharmacology and Physiology. December 27, 2016
    This study describes the pharmacodynamic interaction between propofol and remifentanil. Sixty patients who were scheduled for elective surgery under general anaesthesia (30 males/30 females) were enrolled. Patients were randomly allocated to receive one of 15 combinations of drug levels. Baseline electroencephalograms (EEGs) were recorded for 5 minutes prior to administering the drugs. Patients received a target‐controlled infusion at one of four predefined doses of propofol (high, 3 μg/mL; medium, 1.5 μg/mL; low, 0.5 μg/mL; or no drug) and of remifentanil (high, 6 or 8 ng/mL; medium, 4 ng/mL; low, 2 ng/mL; or no drug). The occurrence of muscle rigidity, apnoea, and loss of consciousness (LOC) was monitored, and EEGs were recorded during the drug administration phase. Electroencephalographic approximate entropy (ApEn) and temporal linear mode complexity (TLMC) parameters at baseline and under steady state conditions were calculated off‐line. Response surfaces were developed to map the interaction between propofol and remifentanil to the probability of occurrence for quantal responses (muscle rigidity, apnoea, LOC) and ApEn and TLMC measurements. Model parameters were estimated using non‐linear mixed effects modelling. The response surface revealed infra‐additive and synergistic effects for muscle rigidity and apnoea, respectively. The effects of the combined drugs on LOC and EEG parameters (eg, ApEn and TLMC) were additive. The C50 estimates of remifentanil (ng/mL) and propofol (μg/mL) were 9.11 and 130 000 for muscle rigidity, 8.99 and 6.26 for apnoea, 13.9 and 3.04 for LOC, 23.4 and 10.4 for ApEn, and 14.8 and 6.51 for TLMC, respectively. The probability of occurrence for muscle rigidity declined when propofol was combined with remifentanil.
    December 27, 2016   doi: 10.1111/1440-1681.12677   open full text
  • Carnosic acid as a component of rosemary extract stimulates skeletal muscle cell glucose uptake via AMPK activation.
    Madina Naimi, Filip Vlavcheski, Brennan Murphy, Tomas Hudlicky, Evangelia Tsiani.
    Clinical and Experimental Pharmacology and Physiology. December 27, 2016
    Compounds that increase the activity of the energy sensor AMP‐activated kinase (AMPK) have the potential to regulate blood glucose levels. Although rosemary extract (RE) has been reported to activate AMPK and reduce blood glucose levels in vivo, the chemical components responsible for these effects are not known. In the present study, we measured the levels of the polyphenol carnosic acid (CA) in RE and examined the effects and the mechanism of action of CA on glucose transport system in muscle cells. High performance liquid chromatography (HPLC) was used to measure the levels of CA in RE. Parental and GLUT4myc or GLUT1myc overexpressing L6 rat myotubes were used. Glucose uptake was assessed using [3H]‐2‐deoxy‐d‐glucose. Total and phosphorylated levels of Akt and AMPK were measured by immunoblotting. Plasma membrane GLUT4myc and GLUT1myc levels were examined using a GLUT translocation assay. Statistics included analysis of variance (ANOVA) followed by Tukey's post‐hoc test. At concentrations found in rosemary extract, CA stimulated glucose uptake in L6 myotubes. At 2.0 μmol/L CA a response (226 ± 9.62% of control, P=.001), similar to maximum insulin (201 ± 7.86% of control, P=.001) and metformin (213 ± 10.74% of control, P=.001) was seen. Akt phosphorylation was not affected by CA while AMPK and ACC phosphorylation was increased and the CA‐stimulated glucose uptake was significantly reduced by the AMPK inhibitor compound C. Plasma membrane GLUT4 or GLUT1 glucose transporter levels were not affected by CA. Our study shows increased muscle cell glucose uptake and AMPK activation by low CA concentrations, found in rosemary extract, indicating that CA may be responsible for the antihyperglycemic properties of rosemary extract seen in vivo.
    December 27, 2016   doi: 10.1111/1440-1681.12674   open full text
  • Melatonin exacerbates acute experimental autoimmune encephalomyelitis by enhancing the serum levels of lactate: A potential biomarker of multiple sclerosis progression.
    Majid Ghareghani, Shima Dokoohaki, Amir Ghanbari, Naser Farhadi, Kazem Zibara, Saeid Khodadoust, Mohammad Parishani, Mehdi Ghavamizadeh, Heibatollah Sadeghi.
    Clinical and Experimental Pharmacology and Physiology. December 27, 2016
    Melatonin has a beneficial role in adult rat models of multiple sclerosis (MS). In this study, melatonin treatment (10 mg/kg/d) was investigated in young age (5‐6 weeks old) Lewis rat model of acute experimental autoimmune encephalomyelitis (EAE) followed by assessing serum levels of lactate and melatonin. Results showed that clinical outcomes were exacerbated in melatonin‐ (neurological score = 6) vs PBS‐treated EAE rats (score = 5). Melatonin caused a significant increase in serum IFN‐γ, in comparison to PBS‐treated EAE rats whereas no considerable change in IL‐4 levels were found, although they were significantly lower than those of controls. The ratio of IFN‐γ/IL‐4, an indicator of Th‐1/Th‐2, was significantly higher in PBS‐ and melatonin‐ treated EAE rats, in comparison to controls. Moreover, results showed increased lymphocyte infiltration, activated astrocytes (GFAP+ cells) but also higher demyelinated plaques (MBP‐deficient areas) in the lumbar spinal cord of melatonin‐treated EAE rats. Finally, serum levels of lactate, but not melatonin, significantly increased in the melatonin group, compared to untreated EAE and normal rats. In conclusion, our results indicated a relationship between age and the development of EAE since a negative impact was found for melatonin on EAE recovery of young rats by enhancing IFN‐γ, the ratio of Th1/Th2 cells, and astrocyte activation, which seems to delay the remyelination process. While melatonin levels decline in MS patients, lactate might be a potential diagnostic biomarker for prediction of disease progression. Early administration of melatonin in the acute phase of MS might be harmful and needs further investigations.
    December 27, 2016   doi: 10.1111/1440-1681.12678   open full text
  • Bone metabolism markers: Indicators of loading dose intravenous ibandronate treatment for bone metastases from breast cancer.
    Ruliang Wang, Shaohua Zhang, Zefei Jiang, Jizheng Tian, Tao Wang, Santai Song.
    Clinical and Experimental Pharmacology and Physiology. December 27, 2016
    To investigate the changes in bone metabolism markers after second‐line treatment with loading dose intravenous (i.v.) ibandronate in patients with bone metastases (BM) from breast cancer, 80 patients were enrolled in this study during January 2010 to April 2014. All the patients were treated with a second‐line loading dose ibandronate for advanced breast cancer with BM and moderate‐to‐severe bone pain. Ibandronate (6 mg) was intravenously administered on three consecutive days followed by maintenance treatment every 3‐4 weeks. Clinical data, including pain score, Karnofsky performance status (KPS) score, and changes in bone metabolism markers, were analyzed. Sixty‐two patients were included in the final analysis. Compared with their pre‐treatment scores, patients exhibited significantly increased KPS scores (P < .01) and a reduced dose of analgesic medication (oxycodone) (P < .01) after 3 and 6 weeks’ post‐treatment. The levels of serum bone alkaline phosphatase (BAP), tartrate‐resistant acid phosphatase (TRACP‐5b), and cross‐linked carboxy‐terminal telopeptide of type I collagen (ICTP) were significantly reduced after 3 and 6 weeks’ post‐treatment (P < .001). Aside from a few adverse events, no liver or renal toxicity was observed. Bone metabolism markers decreased by varying degrees after treatment with a loading dose of ibandronate in patients with BM from breast cancer. It might be convenient using bone metabolism markers to potentially evaluate the efficacy of bisphosphonates treatment for bone metastasis.
    December 27, 2016   doi: 10.1111/1440-1681.12673   open full text
  • c‐Met inhibition enhances chemosensitivity of human ovarian cancer cells.
    Jing Wang, Jian‐Xin Cheng.
    Clinical and Experimental Pharmacology and Physiology. December 27, 2016
    In clinical practice, human ovarian cancer shows considerable resistance to chemotherapy. This study aimed to investigate the role of c‐Met in the chemoresistance of ovarian cancer. Ovarian cancer cell line SKOV3 and OVCAR‐3 were pretreated with c‐Met inhibitor INCB28060, and then treated with paclitaxel. Cell survival, cell cycle and apoptosis were analyzed by MTT assay, flow cytometry analysis and TUNEL assay, respectively. The activation of c‐Met signalling was detected by western blot analysis. INCB28060 inhibited the survival of SKOV3 and OVCAR‐3 cells and enhanced the chemosensitivity of SKOV3 and OVCAR‐3 cells to paclitaxel. INCB28060 inhibited c‐Met signalling, caused mitochondrial membrane depolarization and DNA repair, and induced the apoptosis of SKOV3 and OVCAR‐3 cells. c‐Met plays an important role in mediating the chemoresistance of ovarian cancer. The combination of c‐Met inhibitor and chemotherapy is a promising strategy to human ovarian cancer.
    December 27, 2016   doi: 10.1111/1440-1681.12672   open full text
  • Poly(ADP‐ribose) polymerase 1 deficiency increases nitric oxide production and attenuates aortic atherogenesis through downregulation of arginase II.
    Shu‐jian Wei, Lin Cheng, Er‐shun Liang, Qi Wang, Sheng‐nan Zhou, Hao Xu, Long‐hua Hui, Zhi‐ming Ge, Ming‐xiang Zhang.
    Clinical and Experimental Pharmacology and Physiology. December 27, 2016
    Poly (ADP‐ribose) polymerase (PARP) plays an important role in endothelial dysfunction, leading to atherogenesis and vascular‐related diseases. However, whether PARP regulates nitric oxide (NO), a key regulator of endothelial function, is unclear so far. We investigated whether inhibition of PARP‐1, the most abundant PARP isoform, prevents atherogenesis by regulating NO production and tried to elucidate the possible mechanisms involved in this phenomenon. In apolipoprotein E‐deficient (apoE−/−) mice fed a high‐cholesterol diet for 12 weeks, PARP‐1 inhibition via treatment with 3,4‐dihydro‐54‐(1‐piperindinyl) butoxy‐1(2H)‐isoquinoline (DPQ) or PARP‐1 gene knockout reduced aortic atherosclerotic plaque areas (49% and 46%, respectively). Both the groups showed restored NO production in mouse aortas with reduced arginase II (Arg II) expression compared to that in the controls. In mouse peritoneal macrophages and aortic endothelial cells (MAECs), PARP‐1 knockout resulted in lowered Arg II expression. Moreover, phosphorylation of endothelial NO synthase (eNOS) was preserved in the aortas and MAECs when PARP‐1 was inhibited. Reduced NO production in vitro due to PARP‐1 deficiency could be restored by treating the MAECs with oxidized low‐density lipoprotein treatment, but this effect could not be achieved with peritoneal macrophages, which was likely due to a reduction in the expression of induced NOS expression. Our findings indicate that PARP‐1 inhibition may attenuate atherogenesis by restoring NO production in endothelial cells and thus by reducing Arg II expression and consequently arginase the activity.
    December 27, 2016   doi: 10.1111/1440-1681.12685   open full text
  • Regulation of microvascular flow and metabolism: An overview.
    Michelle A Keske, Renee M Dwyer, Ryan D Russell, Sarah J Blackwood, Aascha A Brown, Donghua Hu, Dino Premilovac, Stephen M Richards, Stephen Rattigan.
    Clinical and Experimental Pharmacology and Physiology. December 27, 2016
    Skeletal muscle is an important site for insulin to regulate blood glucose levels. It is estimated that skeletal muscle is responsible for ~80% of insulin‐mediated glucose disposal in the post‐prandial period. The classical action of insulin to increase muscle glucose uptake involves insulin binding to insulin receptors on myocytes to stimulate glucose transporter 4 (GLUT 4) translocation to the cell surface membrane, enhancing glucose uptake. However, an additional role of insulin that is often under‐appreciated is its action to increase muscle perfusion thereby improving insulin and glucose delivery to myocytes. Either of these responses (myocyte and/or vascular) may be impaired in insulin resistance, and both impairments are apparent in type 2 diabetes, resulting in diminished glucose disposal by muscle. The aim of this review is to report on the growing body of literature suggesting that insulin‐mediated control of skeletal muscle perfusion is an important regulator of muscle glucose uptake and that impairment of microvascular insulin action has important physiological consequences early in the pathogenesis of insulin resistance. This work was discussed at the 2015 Australian Physiological Society Symposium “Physiological mechanisms controlling microvascular flow and muscle metabolism”.
    December 27, 2016   doi: 10.1111/1440-1681.12688   open full text
  • Troxerutin attenuates diet‐induced oxidative stress, impairment of mitochondrial biogenesis and respiratory chain complexes in mice heart.
    Geetha Rajagopalan, Sathiya Priya Chandrasekaran, Anuradha Carani Venkatraman.
    Clinical and Experimental Pharmacology and Physiology. December 27, 2016
    Mitochondrial abnormality is thought to play a key role in cardiac disease originating from the metabolic syndrome (MS). We evaluated the effect of troxerutin (TX), a semi‐synthetic derivative of the natural bioflavanoid rutin, on the respiratory chain complex activity, oxidative stress, mitochondrial biogenesis and dynamics in heart of high fat, high fructose diet (HFFD) ‐induced mouse model of MS. Adult male Mus musculus mice of body weight 25‐30 g were fed either control diet or HFFD for 60 days. Mice from each dietary regimen were divided into two groups on the 16th day and were treated or untreated with TX (150 mg/kg body weight [bw], per oral) for the next 45 days. At the end of experimental period, respiratory chain complex activity, uncoupling proteins (UCP)‐2 and ‐3, mtDNA content, mitochondrial biogenesis and dynamics, oxidative stress markers and reactive oxygen species (ROS) generation were analyzed. Reduced mtDNA abundance with alterations in the expression of genes related to mitochondrial biogenesis and fission and fusion processes were observed in HFFD‐fed mice. Disorganized and smaller mitochondria, reduction in complexes I, III and IV activities (by about 55%) and protein levels of UCP‐2 (52%) and UCP‐3 (46%) were noted in these mice. TX administration suppressed oxidative stress, improved the oxidative capacity and biogenesis and restored fission/fusion imbalance in the cardiac mitochondria of HFFD‐fed mice. TX protects the myocardium by modulating the putative molecules of mitochondrial biogenesis and dynamics and by its anti‐oxidant function in a mouse model of MS.
    December 27, 2016   doi: 10.1111/1440-1681.12671   open full text
  • Smart bomb AS1411 aptamer‐functionalized/PAMAM dendrimer nanocarriers for targeted drug delivery in the treatment of gastric cancer.
    Amir Barzegar Behrooz, Fatemeh Nabavizadeh, Jamal Adiban, Mehdi Shafiee Ardestani, Rouhollah Vahabpour, Mohammad Reza Aghasadeghi, Hamid Sohanaki.
    Clinical and Experimental Pharmacology and Physiology. December 27, 2016
    Chemotherapy, a conventional method assessed in recent oncology studies, poses numerous problems in the clinical environment. To overcome the problems inherent in chemotherapy, an intelligent drug delivery system has come to the forefront of cancer therapeutics. In this study, we designed a dendrimer‐based pharmaceutical system together with a single‐stranded AS1411 aptamer (APTAS1411) as a therapeutic strategy. The polyamidoamine (PAMAM)‐polyethylene glycol (PEG) complex was then conjugated with the AS1411 aptamer and confirmed by atomic‐force microscopy (AFM) and Fourier transform infrared spectroscopy (FTIR) .In this study, we show that the conjugated PAMAM‐PEG‐APTAS1411complex dramatically increased PAMAM‐PEG‐5‐FU uptake by MKN45 gastric cancer cells. We also demonstrated both the stability of the nanoparticle‐5‐FU‐APTAS1411 complex, by thin layer chromatography (TLC), and an increase in 5‐fluorouracil (5‐FU) accumulation in the vicinity of cancerous tumors. This smart drug delivery system is capable of effectively transferring 5‐FU to MKN45 gastric cancer cells in consistent and without toxic effects.
    December 27, 2016   doi: 10.1111/1440-1681.12670   open full text
  • The emerging role of calmodulin regulation of RyR2 in controlling heart rhythm, the progression of heart failure and the antiarrhythmic action of dantrolene.
    Kafa Walweel, Ye Win Oo, Derek R Laver.
    Clinical and Experimental Pharmacology and Physiology. December 27, 2016
    Cardiac output and rhythm depend on the release and the take‐up of calcium from the sarcoplasmic reticulum (SR). Excessive diastolic calcium leak from the SR due to dysfunctional calcium release channels (RyR2) contributes to the formation of delayed after‐depolarizations, which underlie the fatal arrhythmias that occur in heart failure and inherited syndromes. Calmodulin (CaM) is a calcium‐binding protein that regulates target proteins and acts as a calcium sensor. CaM is comprised of two calcium‐binding EF‐hand domains and a flexible linker. CaM is an accessory protein that partially inhibits RyR2 channel activity. CaM is critical for normal cardiac function, and altered CaM binding and efficacy may contribute to defects in SR calcium release. The present paper reviews CaM binding to RyR2 and how it regulates RyR2 channel activity. It then goes on to review how mutations in the CaM amino acid sequence give rise to inherited syndromes such as Catecholaminergic Polymorphic Ventricular Tachychardia (CPVT) and long QT syndrome (LQTS). In addition, the role of reduced CaM binding to RyR2 that results from RyR2 phosphorylation or from oxidation of either RyR2 or CaM contributes to the progression of heart failure is reviewed. Finally, this manuscript reviews recent evidence that CaM binding to RyR2 is required for the inhibitory action of a pharmaceutical agent (dantrolene) on RyR2. Dantrolene is a clinically used muscle relaxant that has recently been found to exert antiarrhythmic effects against SR Ca2+ overload arrhythmias.
    December 27, 2016   doi: 10.1111/1440-1681.12669   open full text
  • l‐arginine and l‐NMMA for assessing cerebral endothelial dysfunction in ischaemic cerebrovascular disease: A systematic review.
    William K Karlsson, Caspar G Sørensen, Christina Kruuse.
    Clinical and Experimental Pharmacology and Physiology. December 27, 2016
    Endothelial dysfunction (ED), in particular cerebral ED, may be an essential biomarker for ischaemic cerebrovascular disease. However, there is no consensus on methods to best estimate cerebral ED. In this systematic review, we evaluate the use of l‐arginine and NG‐monomethyl‐l‐arginine (l‐NMMA) for assessment of cerebral ED. A systematic search of PubMed, EMBASE and the Cochrane Library was done. We included studies investigating cerebrovascular response to l‐arginine or l‐NMMA in human subjects with vascular risk factors or ischaemic cerebrovascular disease. Seven studies (315 subjects) were eligible according to inclusion and exclusion criteria. Studies investigated the effect of age (n=2), type 2 diabetes mellitus (DM) (n=1), cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) (n=1), leukoaraiosis (n=1), and prior ischaemic stroke or transient ischaemic attack (TIA) (n=2) on cerebral ED. Most studies applied transcranial Doppler to quantify cerebral ED. Endothelium‐dependent vasodilatation (EDV) induced by l‐arginine was impaired in elderly and subjects with leukoaraiosis, but enhanced in CADASIL patients. Studies including subjects with prior ischaemic stroke or TIA reported both enhanced and impaired EDV to l‐arginine. Responses to l‐NMMA deviated between subjects with type 2 DM and the elderly. We found only few studies investigating cerebral endothelial responses to l‐arginine and l‐NMMA in subjects with vascular risk factors or ischaemic cerebrovascular disease. Inconsistencies in results were most likely due to variations in methods and included subject populations. In order to use cerebral ED as a prognostic marker, further studies are required to evaluate the association to cerebrovascular disease.
    December 27, 2016   doi: 10.1111/1440-1681.12679   open full text
  • Core skeletal muscle ryanodine receptor calcium release complex.
    Angela F Dulhunty, Lan Wei‐LaPierre, Marco G Casarotto, Nicole A Beard.
    Clinical and Experimental Pharmacology and Physiology. December 27, 2016
    The core skeletal muscle ryanodine receptor (RyR1) calcium release complex extends through three compartments of the muscle fibre, linking the extracellular environment through the cytoplasmic junctional gap to the lumen of the internal sarcoplasmic reticulum (SR) calcium store. The protein complex is essential for skeletal excitation‐contraction (EC)‐coupling and skeletal muscle function. Its importance is highlighted by perinatal death if any one of the EC‐coupling components are missing and by myopathies associated with mutation of any of the proteins. The proteins essential for EC‐coupling include the DHPR α1S subunit in the transverse tubule membrane, the DHPR β1a subunit in the cytosol and the RyR1 ion channel in the SR membrane. The other core proteins are triadin and junctin and calsequestrin, associated mainly with SR. These SR proteins are not essential for survival but exert structural and functional influences that modify the gain of EC‐coupling and maintain normal muscle function. This review summarises our current knowledge of the individual protein/protein interactions within the core complex and their overall contribution to EC‐coupling. We highlight significant areas that provide a continuing challenge for the field. Additional important components of the Ca2+ release complex, such as FKBP12, calmodulin, S100A1 and Stac3 are identified and reviewed elsewhere.
    December 27, 2016   doi: 10.1111/1440-1681.12676   open full text
  • Protective effect of alpha‐lipoic acid, aerobic or resistance exercise from colitis in second hand smoke exposed young rats.
    Dilek Özbeyli, Ayşe Cansu Berberoglu, Anıl Özen, Oktay Erkan, Yunus Başar, Tunahan Şen, Dilek Akakın, Meral Yüksel, Özgür Kasımay Çakır.
    Clinical and Experimental Pharmacology and Physiology. December 27, 2016
    The role of second hand smoke (SHS) exposure on ulcerative colitis is not known. Our aim was to examine the effects of α‐lipoic acid (ALA), chronic aerobic (AE) or resistance exercise (RE) on SHS exposed rats with colitis. Sprague‐Dawley male rats (150‐200 g, n=54) were selected for colitis induction. Among the colitis groups, one group was exposed to SHS (6 d/wk, 4 cigarettes/d) and the other was not. The SHS group was divided into subgroups as follows: sedentary; AE (swimming; 3 d/wk); and RE (climbing with weight; 3 d/wk). After 5 weeks, colitis was induced by intrarectal acetic acid. All groups had subgroups that were given subcutaneously ALA (50 mg/kg per day) or vehicle for 3 days. Following decapitation, colon tissues were sampled to examine malondialdehyde (MDA) and glutathione (GSH) levels, myeloperoxidase (MPO) activity, luminol and lucigenin chemiluminenscence, macroscopic scoring and histologic examination. ANOVA and Student's t‐test were used for statistical analysis. The increased macroscopic and microscopic scores, MPO, MDA, luminol and lucigenin measurements in colitis and SHS‐colitis groups were decreased via ALA (P<.05‐.001). AE declined macroscopic and microscopic scores, MDA, lucigenin compared to colitis and SHS‐colitis groups (P<.01‐.001). RE reduced microscopic score, MPO, MDA, luminol, lucigenin (P<.05‐.001) that were increased with colitis. Decreased GSH levels (P<.01) in the SHS‐colitis group approached to control levels when given ALA. According to our results SHS and colitis induction increased inflammatory damage. SHS did not worsen it more than colitis. Our results suggest that ALA, AE or RE might be protective for SHS exposed ulcerative colitis conditions.
    December 27, 2016   doi: 10.1111/1440-1681.12682   open full text
  • A 12‐week, randomized, parallel‐group, proof‐of‐concept study of tulobuterol patch and salmeterol inhaler as add‐on therapy in adult‐onset mild‐to‐moderate asthma.
    Hideki Inoue, Akio Niimi, Hisako Matsumoto, Isao Ito, Tsuyoshi Oguma, Kojiro Otsuka, Tomoshi Takeda, Hitoshi Nakaji, Tomoko Tajiri, Toshiyuki Iwata, Tadao Nagasaki, Michiaki Mishima.
    Clinical and Experimental Pharmacology and Physiology. December 27, 2016
    Patch formulation of tulobuterol has been used in asthma treatment as a long‐acting β2‐agonist (LABA) through sustained skin absorption. Its treatment efficacy, especially in small airways, remains poorly understood. The study aim was to investigate LABA add‐on effects of tulobuterol patch (TP) and salmeterol inhaler (SA) on pulmonary function, asthma control and health status. Patients who had adult‐onset under‐control asthma, despite taking inhaled corticosteroids, were enrolled in a randomized, open‐label, parallel‐group, proof‐of‐concept study of 12‐week add‐on treatment with TP (n=16) or SA (n=17). Spirometry, impulse oscillometry (IOS), exhaled nitric oxide levels, and clinical questionnaires of asthma control, health status (St. George's Respiratory Questionnaire: SGRQ), and symptoms were evaluated every 4 weeks. Add‐on treatment of SA significantly improved the spirometric indices of small airway obstruction (forced expiratory flow between 25% and 75% of FVC: FEF25‐75, and maximum expiratory flow at 25% of FVC: MEF25) and IOS indices of whole respiratory resistance (resistance at 5 Hz) as compared to TP. In intra‐group comparisons, add‐on treatment of TP improved the scores of the asthma control test and the total SGRQ, as well as the symptom and impact components of the SGRQ. SA add‐on treatment improved FEV1 and IOS parameters of resistance at 20 Hz and reactance at 5 Hz. Neither of the treatments improved exhaled nitric oxide levels. In conclusion, add‐on treatment of TP improved asthma control and health status, whereas SA improved pulmonary function measures associated with large and small airway involvement among patients with adult‐onset mild‐to‐moderate asthma.
    December 27, 2016   doi: 10.1111/1440-1681.12683   open full text
  • Ion channels, long QT syndrome and arrhythmogenesis in ageing.
    Kamalan Jeevaratnam, Karan R Chadda, Samantha C Salvage, Haseeb Valli, Shiraz Ahmad, Andrew A. Grace, Christopher L‐H Huang.
    Clinical and Experimental Pharmacology and Physiology. December 26, 2016
    Ageing is associated with increased prevalences of both atrial and ventricular arrhythmias, reflecting disruption of the normal sequence of ion channel activation and inactivation generating the propagated cardiac action potential. Experimental models with specific ion channel genetic modifications have helped clarify the interacting functional roles of ion channels and how their dysregulation contributes to arrhythmogenic processes at the cellular and systems level. They have also investigated interactions between these ion channel abnormalities and age‐related processes in producing arrhythmic tendency. Previous reviews have explored the relationships between age and loss‐of‐function Nav1.5 mutations in producing arrhythmogenicity. The present review now explores complementary relationships arising from gain‐of‐function Nav1.5 mutations associated with long QT3 (LQTS3). LQTS3 patients show increased risks of life‐threatening ventricular arrhythmias particularly after 40 years of age consistent with such interactions between the ion channel abnormailities and ageing. In turn clinical evidence suggests that ageing is accompanied by structural, particularly fibrotic, as well as electrophysiological change. These abnormalities may result from biochemical changes producing low‐grade inflammation resulting from increased production of reactive oxygen species and superoxide. Experimental studies offer further insights into the underlying mechanisms underlying these phenotypes. Thus, studies in genetically modified murine models for LQTS implicated action potential recovery processes in arrhythmogenesis resulting from functional ion channel abnormalities. In addition, ageing WT murine models demonstrated both ion channel alterations and fibrotic changes with ageing. Murine models then suggested evidence for interactions between ageing and ion channel mutations and provided insights into potential arrhythmic mechanisms inviting future exploration. This article is protected by copyright. All rights reserved.
    December 26, 2016   doi: 10.1111/1440-1681.12721   open full text
  • Inhibition of CD8+ T cells and elimination of myeloid cells by CD4+ Foxp3− T regulatory type 1 cells in acute respiratory distress syndrome.
    Guang‐Gang Li, Ying‐Hua Cao, Yue Run, Ru‐Xiang Xu, Zhen‐Dong Zheng.
    Clinical and Experimental Pharmacology and Physiology. November 21, 2016
    Acute lung injury and acute respiratory distress syndrome (ARDS) are caused by rapid‐onset bilateral pulmonary inflammation. We therefore investigated the potential role of interleukin (IL)‐10+CD4+ Tr1 cells, a regulatory T cell subset with previously identified immunosuppressive functions, in ARDS patients. We first showed that circulating Tr1 cells were upregulated in active and resolved ARDS patients compared to healthy controls and pneumonia patient controls. A significant fraction of these Tr1 cells expressed granzyme B and perforin, while most Tr1 cells did not express interferon gamma (IFN‐γ), IL‐4, IL‐17 or FOXP3, suggesting that the effector functions of these Tr1 cells were primarily mediated by IL‐10, granzyme B, and perforin. Indeed, Tr1 cells effectively suppressed CD8+ T cell IFN‐γ production and induced lysis of monocytes and dendritic cells in vitro. The elimination of myeloid antigen‐presenting cells depended on granzyme B production. We also discovered that Tr1 cells could be identified in the bronchoalveolar lavage fluid collected from ARDS patients. All these results suggested that Tr1 cells possessed the capacity to downregulate inflammation in ARDS. In support of this, we found that ARDS patients who resolved the inflammation and survived the syndrome contained significantly higher levels of Tr1 cells than ARDS patients who succumbed to the syndrome. Overall, this report added a novel piece of evidence that ARDS could be intervened by regulatory T cell‐mediated suppressive mechanisms.
    November 21, 2016   doi: 10.1111/1440-1681.12656   open full text
  • Unfolded protein response is activated in the ipsilateral thalamus following focal cerebral infarction in hypertensive rats.
    Jian Zhang, Hongbing Chen, Weixian Huang, Chunyan Zhou, Jingjing Li, Shihui Xing, Li Chen, Chuo Li, Chao Dang, Gang Liu, Zhong Pei, Jinsheng Zeng.
    Clinical and Experimental Pharmacology and Physiology. November 21, 2016
    Focal cerebral cortical infarction causes secondary neurodegeneration in the remote regions, such as the ventroposterior nucleus of the thalamus. Retrograde degeneration of thalamocortical fibers is considered as the principle mechanism, but the exact molecular events remain to be elucidated. This study aimed to investigate whether unfolded protein response (UPR) is activated in thalamic neurons following distal middle cerebral artery occlusion (MCAO) in stroke‐prone renovascular hypertensive rats. Immunostaining and immunoblotting were performed to evaluate the expression of Grp78 and its downstream effectors in the thalamus at 3, 7 and 14 days after MCAO. Secondary thalamic degeneration was assessed with Nissl staining and NeuN immunostaining. Neuronal death was not apparent at 3 days post‐ischaemia but was evident in the thalamus at 7 and 14 days after MCAO. Grp78 level was reduced in the ipsilateral thalamus at 3 and 7 days after MCAO. In parallel, phosphorylated eIF2α and ATF4 levels were elevated, indicating the activation of UPR. In contrast, ATF6α and CHOP levels were not changed. These results suggest that UPR is activated before neuronal death in the ipsilateral thalamus after MCAO and may represent a key early event in the secondary thalamic degeneration.
    November 21, 2016   doi: 10.1111/1440-1681.12657   open full text
  • Impact of maternal cigarette smoke exposure on brain and kidney health outcomes in female offspring.
    Yik L Chan, Sonia Saad, Ibrahim Al‐Odat, Amgad A Zaky, Brian Oliver, Carol Pollock, Weihong Li, Nicole M Jones, Hui Chen.
    Clinical and Experimental Pharmacology and Physiology. November 21, 2016
    Increased oxidative stress in the brain can lead to increased sympathetic tone that may further induce kidney dysfunction. Previously we have shown that maternal cigarette smoke exposure (SE) leads to significantly increased oxidative stress and inflammation in both brain and kidney, as well as reduced brain and kidney mitochondrial activity. This is closely associated with significant kidney underdevelopment and abnormal function in adulthood in the male offspring. This study aimed to investigate the impact of maternal SE on brain and kidney health in the female offspring. In this study, the mouse dams were exposed to two cigarettes, twice daily for 6 weeks prior to gestation, during pregnancy and lactation. Brains and kidneys from the female offspring were collected at 20 days (P20) and 13 weeks (W13) and were subject to further analysis. We found that mRNA expression of brain inflammatory markers interleukin‐1 receptor and Toll‐like receptor 4 were significantly increased in the SE offspring at both P20 and W13. Their brain mitochondrial activity markers were however increased at W13 with increased antioxidant activity. Kidney development and function in the female SE offspring were not different from the control offspring. We concluded that although brain inflammatory markers were upregulated in the SE female offspring, they were protected from some of the indicators of brain oxidative stress, such as endogenous antioxidant and mitochondrial dysfunction, as well as abnormal kidney development and function in adulthood.
    November 21, 2016   doi: 10.1111/1440-1681.12659   open full text
  • Beneficial effect of aspirin against interferon‐α‐2b—induced depressive behavior in Sprague Dawley rats.
    Shailendra Bhatt, Kilambi Pundarikakshudu, Paresh Patel, Nirav Patel, Ashish Panchal, Gaurang Shah, Sunita Goswami.
    Clinical and Experimental Pharmacology and Physiology. November 21, 2016
    Accumulating data advocates that inflammatory mediators may contribute to depression in experimental models as well as in humans. Nonetheless, whether anti‐inflammatory treatments can prevent depression still remains controversial. To substantiate our hypothesis, we used an interferon‐α‐2b model of depression using Sprague Dawley rats. Interferon‐α‐2b is a cytokine which activates immune response and also produces depression. The animals were treated for 21 days with aspirin (10 mg/kg, per oral (p.o.)) dexamethasone (1 mg/kg p.o.) and amitriptyline (10 mg/kg p.o.). Amitriptyline was used as reference standard, and given concurrently with aspirin and dexamethasone to examine any synergy. Interferon‐α‐2b (6000 IU/kg, intraperitoneal (i.p.)) was administered in all the above groups daily, except normal control. Tests performed included sucrose preference test, behavioural tests like forced swim test, elevated plus maze, light dark box and locomotor activity along with biochemical estimations like serum cortisol and brain neurotransmitters. The rats in the group treated with Interferon‐α‐2b produced depressive behaviour in rats. We found that animals treated with aspirin decreased immobility time in forced swim test, increased sucrose preference, decreased serum cortisol and increased brain serotonin levels signifying antidepressant action. In contrast, there was no effect in groups treated with dexamethasone. Our results suggest that aspirin can serve as a potential antidepressant both individually and as adjuvant agent in the treatment of depression. Inhibition of the cyclo‐oxygenase‐2 levels and prostaglandins concentration or any other potential physiological and biochemical mechanisms may be involved in antidepressant effect.
    November 21, 2016   doi: 10.1111/1440-1681.12660   open full text
  • The effect of continuous positive airway pressure on heart rate variability during the night in patients with chronic heart failure and central sleep apnoea.
    Kiril V Terziyski, Aneliya I Draganova, Zdravko Z Taralov, Ilcho S Ilchev, Stefan S Kostianev.
    Clinical and Experimental Pharmacology and Physiology. November 21, 2016
    Continuous positive airway pressure (CPAP) improves autonomic activity in patients with chronic heart failure (CHF) and central sleep apnoea (CSA), but its effect on heart rate variability (HRV) during therapy has not been reported. We hypothesized that CPAP may decrease HRV, despite its beneficial effects on sympathetic overactivation, due to the expected stabilization of breathing. Sixty‐seven CHF patients underwent polysomnography (PSG). Ten of them presented with CSA (age 66.1±8.5 years, apnoea‐hypopnea index [AHI]=57.6±23.3, central AHI [cAHI]=41.6±24.6 [mean±SD]) and were subjected to a second PSG with manual CPAP titration. Beat‐to‐beat heart intervals for a 6‐hour period of sleep were extracted from each recording and HRV was analysed. CPAP significantly reduced AHI (AHI=23.1±18.3 P=.004). Standard deviation of normal‐normal interbeat interval (SDNN) (61.5±29.0 vs 49.5±19.3 ms, P=.021), root mean square of successive differences (RMSSD) (21.8±9.2 vs 16.4±7.1 ms, P=.042), total power (lnTP=7.8±1.1 vs 7.4±0.8 ms2, P=.037), low frequency power (lnLF=5.5±1.5 vs 5.0±1.4 ms2, P=.003) and high frequency power (lnHF=4.6±1.0 vs 4.0±1.0 ms2, P=.024) were decreased. There was a strong correlation between the decrease in AHI and the decrease in lnHF (Spearman's ρ=.782). CPAP leads to a decrease in spectral and time domain parameters of HRV during therapy in CHF patients with CSA. These changes are best explained by the effect which CPAP‐influenced breathing pattern and lowered AHI exert on HRV.
    November 21, 2016   doi: 10.1111/1440-1681.12662   open full text
  • Juvenile growth reduces the influence of epithelial sodium channels on myogenic tone in skeletal muscle arterioles.
    Lori S Kang, Shyama Masilamani, Matthew A Boegehold.
    Clinical and Experimental Pharmacology and Physiology. November 21, 2016
    Previous studies have documented that rapid juvenile growth is accompanied by functional changes in the arteriolar endothelium, but much less is known about functional changes in arteriolar smooth muscle over this period. In this study, we investigate the possible contribution of epithelial sodium channels (ENaC) to the myogenic behaviour of arterioles at two stages of juvenile growth. The effects of the ENaC inhibitor benzamil on different levels of myogenic tone were studied in isolated gracilis muscle arterioles from rats aged 21‐28 days (“weanlings”) and 42‐49 days (“juveniles”). ENaC subunit expression in the arteriolar wall was also determined, and the interaction between ENaC and nitric oxide (NO) in regulating vascular tone was explored by combined use of benzamil and NG‐monomethyl‐l‐arginine (l‐NMMA). At physiological pressures, both steady‐state myogenic tone and the dynamic adjustments in this tone triggered by acute pressure changes were less in juvenile arterioles than in weanling arterioles. α, β and γ ENaC protein was present in arterioles at both ages, but benzamil only had an effect on myogenic tone in weanling arterioles. In these vessels, benzamil increased, rather than decreased, myogenic tone, and this effect was prevented by l‐NMMA or endothelial removal. These findings suggest that although ENaC is present in gracilis muscle arterioles of both weanling and juvenile rats, it is not obligatory for the genesis of myogenic activity in these vessels at either age. However, ENaC activity can significantly modulate the level of myogenic tone through stimulation of endothelial NO release at an early stage of growth.
    November 21, 2016   doi: 10.1111/1440-1681.12664   open full text
  • Bacopa monnieri (Brahmi) improved novel object recognition task and increased cerebral vesicular glutamate transporter type 3 in sub‐chronic phencyclidine rat model of schizophrenia.
    Pritsana Piyabhan, Supaporn Wannasiri, Jarinyaporn Naowaboot.
    Clinical and Experimental Pharmacology and Physiology. November 21, 2016
    Reduced vesicular glutamate transporter 1 (VGLUT1) and 2 (VGLUT2) indicate glutamatergic hypofunction leading to cognitive impairment in schizophrenia. However, VGLUT3 involvement in cognitive dysfunction has not been reported in schizophrenia. Brahmi (Bacopa monnieri) might be a new treatment and prevention for cognitive deficits in schizophrenia by acting on cerebral VGLUT3 density. We aimed to study cognitive enhancement‐ and neuroprotective‐effects of Brahmi on novel object recognition and cerebral VGLUT3 immunodensity in sub‐chronic (2 mg/kg, Bid, ip) phencyclidine (PCP) rat model of schizophrenia. Rats were assigned to three groups for cognitive enhancement effect study: Group 1, Control; Group 2, PCP administration; Group 3, PCP+Brahmi. A neuroprotective‐effect study was also carried out. Rats were again assigned to three groups: Group 1, Control; Group 2, PCP administration; Group 3, Brahmi+PCP. Discrimination ratio (DR) representing cognitive ability was obtained from a novel object recognition task. VGLUT3 immunodensity was measured in the prefrontal cortex, striatum and cornu ammonis fields 1–3 (CA1–3) using immunohistochemistry. We found reduced DR in the PCP group, which occurred alongside VGLUT3 reduction in all brain areas. PCP+Brahmi showed higher DR score with increased VGLUT3 immunodensity in the prefrontal cortex and striatum. Brahmi+PCP group showed a higher DR score with increased VGLUT3 immunodensity in the prefrontal cortex, striatum and CA1–3. We concluded that reduced cerebral VGLUT3 was involved in cognitive deficit in PCP‐administrated rats. Receiving Brahmi after PCP restored cognitive deficit by increasing VGLUT3 in the prefrontal cortex and striatum. Receiving Brahmi before PCP prevented cognitive impairment by elevating VGLUT3 in prefrontal cortex, striatum and CA1–3. Therefore, Brahmi could be a new frontier of restoration and prevention of cognitive deficit in schizophrenia.
    November 21, 2016   doi: 10.1111/1440-1681.12658   open full text
  • Brattleboro rats have impaired apical membrane water permeability regulation in the outer medullary collecting duct principal cells.
    Galina S Baturina, Liubov E Katkova, Sotirios G Zarogiannis, Evgeniy I Solenov.
    Clinical and Experimental Pharmacology and Physiology. November 21, 2016
    Vasopressin (AVP) regulates the body salt‐water balance. Brattleboro rats carry an AVP gene mutation resulting in a recessive form of central diabetes insipidus, being ideal for AVP deficiency studies. Herein, we studied the water permeability of the apical and basolateral sides of outer medullary collecting duct (OMCD) principal cells in response to dDAVP (a V2 receptor agonist) administration in Wistar and Brattleboro rats. Biophysical measurements of the water permeability (Pf) of isolated OMCD principal cells were performed with the calcein quenching method with/without dDAVP (10−8 mol/L). mRNA transcripts and protein levels of AQP2, AQP3 and AQP4 were assessed by RT‐PCR and western blot respectively. dDAVP increased the apical and basolateral Pf of OMCD principal cells in Wistar rats, while in Brattleboro rats this effect was present basolaterally. Long‐term dDAVP administration in both strains resulted in a significant increase in mRNA expression of all assessed AQP's while only the protein levels of AQP2 and AQP3 were significantly increased. Short‐term (20 minutes) dDAVP treatment of isolated OMCD fragments resulted in significantly increased plasma membrane expression of AQP2 in Wistar rats and of AQP2 and AQP3 in Brattleboro rats. In summary, dDAVP induces different expression of AQP2, AQP3 and AQP4 in Wistar and Brattleboro rats during short‐ and long‐term treatment. In Wistar rats dDAVP mainly increased AQP2 expression while in Brattleboro rats it increased functional water permeability mainly by AQP3 expression.
    November 21, 2016   doi: 10.1111/1440-1681.12666   open full text
  • Maternal protein restriction induced‐hypertension is associated to oxidative disruption at transcriptional and functional levels in the medulla oblongata.
    José L Brito Alves, Jéssica M D Oliveira, Diorginis J S Ferreira, Monique A de V Barros, Viviane O Nogueira, Débora S Alves, Hubert Vidal, Carol G Leandro, Cláudia J Lagranha, Luciano Pirola, João H Costa‐Silva.
    Clinical and Experimental Pharmacology and Physiology. November 21, 2016
    Maternal protein restriction during pregnancy and lactation predisposes the adult offspring to sympathetic overactivity and arterial hypertension. Although the underlying mechanisms are poorly understood, dysregulation of the oxidative balance has been proposed as a putative trigger of neural‐induced hypertension. The aim of the study was to evaluate the association between the oxidative status at transcriptional and functional levels in the medulla oblongata and maternal protein restriction induced‐hypertension. Wistar rat dams were fed a control (normal protein; 17% protein) or a low protein ((Lp); 8% protein) diet during pregnancy and lactation, and male offspring was studied at 90 days of age. Direct measurements of baseline arterial blood pressure (ABP) and heart rate (HR) were recorded in awakened offspring. In addition, quantitative RT‐PCR was used to assess the mRNA expression of superoxide dismutase 1 (SOD1) and 2 (SOD2), catalase (CAT), glutathione peroxidase (GPx), Glutamatergic receptors (Grin1, Gria1 and Grm1) and GABA(A)‐receptor‐associated protein like 1 (Gabarapl1). Malondialdehyde (MDA) levels, CAT and SOD activities were examined in ventral and dorsal medulla. Lp rats exhibited higher ABP. The mRNA expression levels of SOD2, GPx and Gabarapl1 were down regulated in medullary tissue of Lp rats (P<.05, t test). In addition, we observed that higher MDA levels were associated to decreased SOD (approximately 45%) and CAT (approximately 50%) activities in ventral medulla. Taken together, our data suggest that maternal protein restriction induced‐hypertension is associated with medullary oxidative dysfunction at transcriptional level and with impaired antioxidant capacity in the ventral medulla.
    November 21, 2016   doi: 10.1111/1440-1681.12667   open full text
  • Novel cardioprotection strategies for the aged heart: evidence from pre‐clinical studies.
    Alice E Kane, Susan E Howlett.
    Clinical and Experimental Pharmacology and Physiology. November 21, 2016
    The incidence of cardiovascular disease is rising as the population ages. This has led to an increase in the need to perform cardiac surgery in older patients. However, aged hearts are particularly susceptible to reperfusion injury following periods of myocardial ischaemia that occur during cardiac surgery. Indeed, older adults experience myocardial dysfunction and reduced survival post‐surgery compared to younger people and certain groups, including older women and frail older adults, are at particular risk. This highlights the need to design cardioprotective strategies specifically for the ageing heart. Cardioprotection during surgery is often accomplished by perfusing the heart with chemical arresting agents, known as cardioplegic solutions. New protective strategies have been developed and tested in animal models, where cardioplegic solutions have been modified by changing their temperature, chemical components and/or the frequency of delivery. In addition, drugs designed to activate cardioprotective mechanisms or to inhibit mechanisms involved in injury have been added to improve the efficacy of these solutions. However, most experimental studies have developed and optimized cardioplegic solutions in hearts from younger male animals. This review discusses pre‐clinical models used to optimize cardioplegic solutions, with an emphasis on the few studies that have used hearts from older animals. Pharmacologic agents that have been shown to enhance the benefits of cardioplegia in younger hearts and could, in theory, protect vulnerable older hearts are also considered. We emphasize the need to conduct studies in frail older animals of both sexes to facilitate translation of laboratory‐based observations to the clinic.
    November 21, 2016   doi: 10.1111/1440-1681.12668   open full text
  • Epigallocatechin‐3‐gallate augments therapeutic effects of mesenchymal stem cells in skin wound healing.
    Min Li, Jingxing Xu, Tongxin Shi, Haiyang Yu, Jianping Bi, Guanzhi Chen.
    Clinical and Experimental Pharmacology and Physiology. October 20, 2016
    In non‐healing wounds, mesenchymal stem cell (MSC)‐based therapies have the potential to activate a series of coordinated cellular processes, including angiogenesis, inflammation, cell migration, proliferation and epidermal terminal differentiation. As pro‐inflammatory reactions play indispensable roles in initiating wound repair, sustained and prolonged inflammation exhibit detrimental effects on skin wound closure. We investigated the feasibility of using an antioxidant agent epigallocatechin‐3‐gallate (EGCG), along with MSCs, to improve wound repair through their immunomodulatory actions. In a rat model of wound healing, a single dose of EGCG at 10 mg/kg increased the efficiency of MSC‐induced skin wound closure. Twenty days after the wound induction, MSC treatment significantly enhanced the epidermal thickness, which was further increased by EGCG administration. Consistently, the highest extent of growth factors upregulation for neovascularization induction was seen in the animals treated by both MSCs and EGCG, associated with a potent anti‐scarring effect throughout the healing process. Finally, expression levels of pro‐inflammatory cytokines, such as tumor necrosis factor‐α (TNF‐α), interleukin‐1β (IL‐1β) and IL‐6, in the wound area were reduced by MSCs, and this reduction was further potentiated by EGCG co‐administration. EGCG, together with MSCs, can promote skin wound healing likely through their combinational effects in modulating chronic inflammation.
    October 20, 2016   doi: 10.1111/1440-1681.12652   open full text
  • Moderate traumatic brain injury is linked to acute behaviour deficits and long term mitochondrial alterations.
    Hui Chen, Yik Lung Chan, Long The Nguyen, Yilin Mao, Alicia de Rosa, Ing Tsyr Beh, Candice Chee, Brian Oliver, George Herok, Sonia Saad, Catherine Gorrie.
    Clinical and Experimental Pharmacology and Physiology. October 20, 2016
    Traumatic brain injury (TBI) remains one of the leading causes of death and disability worldwide. Mild TBI may lead to neuropsychiatric sequelae, including memory loss and motor impairment. Mitochondrial dysfunction and oxidative stress have a contributory role in several neurological disorders; however, their association with mitophagy in mild TBI is unclear. TBI was induced in female Sprague Dawley (SD) rats using a New York University Impactor (10 g, impactor head 2.5 mm diameter, weight drop 50 mm) and compared to sham surgery controls. The novel object recognition and error ladder tests were performed at 24 hours and for 6 weeks post injury, and the brains were examined histologically to confirm the extent of injury. Mitochondria manganese superoxide dismutase (MnSOD) and the oxidative phosphorylation (OXPHOS) complexes I‐V (CI‐CV), as well as mitophagy markers, dynamin related protein 1 (DRP‐1), LC3A/B and PTEN‐induced putative kinase 1 (PINK‐1), were measured in the penumbra by western blot. At 24 hours sham rats performed as expected on a novel object recognition test while TBI rats showed cognitive deficits at the early time points. TBI rats also showed more early motor deficits on a horizontal ladder, compared with the sham rats. MnSOD, OXPHOS CI, CIII and CV protein levels were significantly lower in the TBI group at 24 hours. DRP‐1, LC3A/B I and II, and PINK‐1 were increased at 6 weeks suggesting abnormal mitophagy. Moderate TBI caused immediate cognitive and mild motor functional deficits in the rats that did not persist. Reduced antioxidative capacity and possibly compromised mitochondrial function may affect the long term functional recovery.
    October 20, 2016   doi: 10.1111/1440-1681.12650   open full text
  • Vasorelaxant effects of 2‐nitro‐1‐phenyl‐1‐propanol in rat aorta.
    Teresinha Silva Brito, Francisco José Batista‐Lima, Kalinne Kelly Lima Gadelha, Patrícia Andrea Fonseca‐Magalhães, Saad Lahlou, Pedro Jorge Caldas Magalhães.
    Clinical and Experimental Pharmacology and Physiology. October 20, 2016
    2‐Nitro‐1‐phenyl‐1‐propanol (NPP) is a nitro alcohol that is known as an intermediate in the synthesis of sympathomimetic agents, such as norephedrine. The present study investigated the vasoactive effects of NPP on rat aorta. In endothelium‐intact aortic rings, NPP fully relaxed contractions that were induced by phenylephrine, KCl, and U‐46619. The relaxant effects of NPP on phenylephrine‐elicited contractions remained unaffected by NG‐nitro‐l‐arginine methyl ester (l‐NAME), indomethacin, propranolol, tetraethylammonium, 4‐aminopyridine, and glibenclamide. Conversely, pretreatment with 1H‐[1,2,4]oxadiazolo[4,3‐a]quinoxalin‐1‐one (ODQ), cis‐N‐(2‐phenylcyclopentyl)‐azacyclotridec‐1‐en‐2‐amine hydrochloride (MDL‐12,330A), and N‐[2‐(P‐bromocinnamylamino)ethyl]‐5‐isoquinolinesulfonamide dihydrochloride (H‐89) reduced the ability of NPP to relax contractions that were elicited by phenylephrine. NPP inhibited the vasoconstrictor response that was induced by Ca2+ in aortic rings that were stimulated by pharmacomechanical or electromechanical coupling with phenylephrine and 60 mmol/L KCl, respectively, and after the depletion of intracellular Ca2+ stores. Such effects of NPP were significantly reversed by pretreatment with the guanylyl cyclase inhibitor ODQ and weakly influenced by the adenylyl cyclase inhibitor MDL‐12,330A. In Ca2+‐free medium, NPP inhibited transient contractions that were induced by phenylephrine but not caffeine. In homogenates of aortic rings, NPP increased cyclic guanosine 3′,5′‐monophosphate (cGMP) and cyclic adenosine 3′‐5′‐monophosphate levels, but this effect was statistically significant only for cGMP. In conclusion, in contrast to the vasoconstrictor amine norephedrine, NPP is a vasodilator in rat aorta, and its relaxant effects are likely attributable to cGMP production.
    October 20, 2016   doi: 10.1111/1440-1681.12625   open full text
  • Evaluations of lipid peroxidation and inflammation in short‐term glycerol‐induced acute kidney injury in rats.
    Akina Nara, Daisuke Yajima, Sayaka Nagasawa, Hiroko Abe, Yumi Hoshioka, Hirotaro Iwase.
    Clinical and Experimental Pharmacology and Physiology. October 20, 2016
    Rhabdomyolysis is characterised by acute kidney injury (AKI) resulting from skeletal muscle injury. Lipid peroxidation‐mediated oxidant injury and pro‐inflammatory cytokine‐mediated inflammatory response play critical roles in the pathogenesis of rhabdomyolysis‐induced AKI. The present study aimed to investigate the short‐term effects of both lipid peroxidation and inflammatory responses on rhabdomyolysis‐induced AKI in a rat model of glycerol‐induced rhabdomyolysis. Rhabdomyolysis was induced by the intramuscular injection of 50% glycerol in saline (10 mL/kg) into the hind limbs of rats. Rats were killed 1 or 3 hours after glycerol injection. Time‐dependent increases in serum biochemical parameters, including blood urea nitrogen, creatinine, lactate dehydrogenase and creatine phosphokinase levels, were observed 1 hour after glycerol injection. In kidneys, glycerol injection resulted in histopathological changes such as renal tubular injury and renal tubular myoglobin deposition. Levels of Nε‐(hexanoyl)lysine‐modified, 4‐hydroxy‐2‐nonenal‐modified, and nitrotyrosine‐modified proteins in rat kidneys were unaltered at 1 hour after glycerol injection, but increased significantly at 3 hours. Increases in renal nitric oxide production and the expression levels of inducible nitric oxide synthase, interleukin‐6 and tumour necrosis factor‐α in the renal parenchyma were observed at 1 hour after glycerol injection and plateaued at 3 hours. Our findings suggest that the pro‐inflammatory cytokine‐mediated inflammatory response may cause rhabdomyolysis‐induced AKI very shortly after glycerol injection, and lipid peroxidation‐mediated oxidant injury may promote the development of these pathophysiological processes.
    October 20, 2016   doi: 10.1111/1440-1681.12633   open full text
  • Inhibition of macrophage migration inhibitory factor (MIF) tautomerase activity suppresses microglia‐mediated inflammatory responses.
    Yu Zhang, Ruinan Gu, Jia Jia, Tingjun Hou, Long Tai Zheng, Xuechu Zhen.
    Clinical and Experimental Pharmacology and Physiology. October 20, 2016
    Macrophage migration inhibitory factor (MIF), a pleiotropic pro‐inflammatory cytokine, is a key regulator in both innate and acquired immunity systems. MIF has become a promising drug target for inflammatory diseases. Apart from its cytokine activities, MIF is known to act as a d‐dopachrome tautomerase. Our previous work has identified that 3‐[(biphenyl‐4‐ylcarbonyl)carbamothioyl]amino benzoic acid (Z‐590) exhibited a potent inhibitory activity against MIF. In this study, we investigate the effect of Z‐590 on lipopolysaccharide (LPS)‐activated microglial cell activation. Our results demonstrate that Z‐590 significantly decreases the production of nitric oxide (NO), tumour necrosis factor‐alpha (TNF‐α), interleukin (IL)‐6, IL‐1β, cyclooxygenase (COX‐2), inducible nitric oxide synthase (iNOS) as well as reactive oxygen species (ROS) involved in inhibiting MAKPs signalling pathway in LPS‐stimulated microglia cells. Furthermore, Z‐590 reduced cytotoxicity of activated microglia toward HT‐22 hippocampal cells in a microglia‐conditioned medium system. Taken together, these results indicate that MIF inhibitor Z‐590 elicits a potent inhibitor for microglia‐mediated neuroinflammation.
    October 20, 2016   doi: 10.1111/1440-1681.12647   open full text
  • Hydrogen peroxide scavenger, catalase, alleviates ion transport dysfunction in murine colitis.
    Kim E Barrett, Declan F McCole.
    Clinical and Experimental Pharmacology and Physiology. October 20, 2016
    Reactive oxygen species (ROS) such as hydrogen peroxide (H2O2) contribute to epithelial damage and ion transport dysfunction (key events in inflammatory diarrhoea) in inflammatory bowel disease (IBD). The aim of this study was to identify if H2O2 mediates suppression of colonic ion transport function in the murine dextran sulfate sodium (DSS) colitis model by using the H2O2 degrading enzyme, catalase. Colitis was induced by administering DSS (4%) in drinking water for 5 days followed by 3 days on normal H2O. Mice were administered either pegylated catalase or saline at day −1, 0 and +1 of DSS treatment. Ion transport responses to the Ca2+‐dependent agonist, carbachol (CCh), or the cAMP‐dependent agonist, forskolin, were measured across distal colonic mucosa mounted in Ussing chambers. Parameters of DSS‐induced inflammation (loss in body weight, decreased colon length, altered stool consistency), were only partially alleviated by catalase while histology was only minimally improved. However, catalase significantly reversed the DSS‐induced reduction in baseline ion transport as well as colonic Isc responses to CCh. However, ion transport responses to forskolin were not significantly restored. Catalase also reduced activation of ERK MAP kinase in the setting of colitis, and increased expression of the Na+‐K+‐2Cl− cotransporter, NKCC1, consistent with restoration of ion transport function. Ex vivo treatment of inflamed colonic mucosae with catalase also partially restored ion transport function. Therefore, catalase partially prevents, and rescues, the loss of ion transport properties in DSS colitis even in the setting of unresolved tissue inflammation. These findings indicate a prominent role for ROS in ion transport dysfunction in colitis and may suggest novel strategies for the treatment of inflammatory diarrhoea.
    October 20, 2016   doi: 10.1111/1440-1681.12646   open full text
  • Emodin ameliorates acute lung injury induced by severe acute pancreatitis through the up‐regulated expressions of AQP1 and AQP5 in lung.
    Junfeng Xu, Bo Huang, Yu Wang, Caiyu Tong, Peng Xie, Rong Fan, Zhenming Gao.
    Clinical and Experimental Pharmacology and Physiology. October 20, 2016
    The present study investigates the ameliorating effects of emodin on acute lung injury (ALI) induced by severe acute pancreatitis (SAP). An ALI rat model was constructed by sodium ursodeoxycholate and they were divided into four groups: SHAM, ALI, emodin and dexamethasone (DEX) (n=24 per group). Blood samples and lung tissues were collected 6, 12 and 24 hours after the induction of SAP‐associated ALI. Lung wet/dry ratio, blood gases, serum amylase and tumor necrosis factor‐α (TNF‐α) were measured at each time point. The expressions of AQP1 and AQP5 in lung tissue were detected by immunohistochemical staining, western blotting and real‐time PCR. As the results show, there were no statistical differences in the levels of serum amylase, lung wet/dry ratio, blood gases indexes, serum TNF‐α and pathological changes between emodin and DEX groups. However, significant differences were observed when compared with the ALI group. AQP1 and AQP5 expressions were significantly increased and lung oedemas were alleviated with the treatment of emodin and DEX. The expressions of AQP1 and AQP5 were significantly decreased in SAP‐associated ALI rats. Emodin up‐regulated the expression of AQP1 and AQP5, it could reduce pulmonary oedema and ameliorate SAP‐induced ALI. Regulations on AQP1 and AQP5 expression had a great value in clinical application.
    October 20, 2016   doi: 10.1111/1440-1681.12627   open full text
  • The CD147/MMP‐2 signaling pathway may regulate early stage cardiac remodelling in spontaneously hypertensive rats.
    Bowei Li, Wanxing Zhou, Xiaorong Yang, Yuliang Zhou, Yongjing Tan, Congcong Yuan, Yulan Song, Xiao Chen, Wei Zhang.
    Clinical and Experimental Pharmacology and Physiology. October 20, 2016
    Previous studies have reported that decreased matrix metalloproteinase‐2 (MMP‐2) is associated with early stage (age 8–16 weeks) ventricular remodelling in spontaneously hypertensive rats (SHR). We hypothesized that inhibited CD147/MMP‐2 signalling might down‐regulate MMP‐2 expression and augment remodelling in spontaneously hypertensive rats. Twenty‐nine male SHR (8 weeks) were randomly assigned to SHR, CD147, and CD147+DOX groups. The control group included eight age‐matched WKY rats. CD147 and CD147+DOX groups received recombinant human CD147 (600 ng/kg in 1.5 mL saline, weekly). The SHR and WKY groups received the vehicle. The CD147+DOX group also received doxycycline, an inhibitor of MMPs (daily, 30 mg/kg in 1.5 mL saline, iG). On day 56 echocardiography and left ventricular mass index (LVWI) measurements were collected and histological sections were stained for cell and collagen content. Myocardium MMP‐2, TIMP‐1, CD147, and collagens types I and III were estimated by western blot. CD147 and the ratio of MMP‐2/TIMP‐1 were lower in SHR than WKY rats (P<.05). Myocyte hypertrophy, partial fibre breaks, plasmolysis, necrosis and collagen content (collagen volume fraction [CVF], I and III) in SHR were above control levels (P<.05). CD147 rats showed CD147, MMP‐2 and MMP‐2/TIMP‐1 were increased (P<.05), CVF, LVWI, and collagen I and III were decreased (P<.05) and myocyte morphology was improved. CD147 levels did not differ between CD147+DOX and CD147 groups, CVF, collagens type I and III and partial fiber breaks were more abundant in CD147+DOX (P<.05). In summary, an inhibited CD147/MMP‐2 pathway was associated with early stage cardiac remodelling, and CD147 supplementation may attenuate this response.
    October 20, 2016   doi: 10.1111/1440-1681.12626   open full text
  • Effects of JTV‐519 on stretch‐induced manifestations of mechanoelectric feedback.
    Irene Canto, Luis Such‐Miquel, Laia Brines, Carlos Soler, Manuel Zarzoso, Conrado Calvo, Germán Parra, Álvaro Tormos, Antonio Alberola, José Millet, Luis Such, Francisco J. Chorro.
    Clinical and Experimental Pharmacology and Physiology. October 20, 2016
    JTV‐519 is a 1,4‐benzothiazepine derivative with multichannel effects that inhibits Ca2+ release from the sarcoplasmic reticulum and stabilizes the closed state of the ryanodine receptor, preventing myocardial damage and the induction of arrhythmias during Ca2+ overload. Mechanical stretch increases cellular Na+ inflow, activates the reverse mode of the Na+/Ca2+ exchanger, and modifies Ca2+ handling and myocardial electrophysiology, favoring arrhythmogenesis. This study aims to determine whether JTV‐519 modifies the stretch‐induced manifestations of mechanoelectric feedback. The ventricular fibrillation (VF) modifications induced by acute stretch were studied in Langendorff‐perfused rabbit hearts using epicardial multiple electrodes under control conditions (n=9) or during JTV‐519 perfusion: 0.1 μmol/L (n=9) and 1 μmol/L (n=9). Spectral and mapping techniques were used to establish the baseline, stretch and post‐stretch VF characteristics. JTV‐519 slowed baseline VF and decreased activation complexity. These effects were dose‐dependent (baseline VF dominant frequency: control=13.9±2.2 Hz; JTV 0.1 μmol/L=11.1±1.1 Hz, P<.01; JTV 1 μmol/L=6.6±1.1 Hz, P<.0001). The stretch‐induced acceleration of VF (control=38.8%) was significantly reduced by JTV‐519 0.1 μmol/L (19.8%) and abolished by JTV 1 μmol/L (−1.5%). During stretch, the VF activation complexity index was reduced in both JTV‐519 series (control=1.60±0.15; JTV 0.1 μmol/L=1.13±0.3, P<.0001; JTV 1 μmol/L=0.57±0.21, P<.0001), and was independently related to VF dominant frequency (R=.82; P<.0001). The fifth percentile of the VF activation intervals, conduction velocity and wavelength entered the multiple linear regression model using dominant frequency as the dependent variable (R=−.84; P<.0001). In conclusion, JTV‐519 slowed and simplified the baseline VF activation patterns and abolished the stretch‐induced manifestations of mechanoelectric feedback.
    October 20, 2016   doi: 10.1111/1440-1681.12630   open full text
  • Hypertension in rat offspring subjected to perinatal protein malnutrition is not related to the baroreflex dysfunction.
    K M Paulino‐Silva, J H Costa‐Silva.
    Clinical and Experimental Pharmacology and Physiology. October 20, 2016
    This study reports on the effects of maternal protein malnutrition on baroreflex (BR) control of the heart rate and sympathetic nerve activity in the hypertensive male offspring of Wistar rat dams. Wistar rat dams were fed a normal protein (NP) control (17% protein) or a low protein (LP; 8% protein) diet during pregnancy and lactation, and their male offspring were studied when 90 days old. In these animals we evaluated spontaneous and induced BR control, the variability of the cardiovascular system and analyzed a direct recording of lumbar sympathetic nervous activity. The 90 day‐old LP conscious rats had increased arterial pressure compared to NP, with enhanced low frequency oscillations of the systolic pressure, but no changes in the spontaneous and induced BR control of heart rate. In relation to nerve recordings, we observed similar values in terms of mean, frequency and amplitude between the groups. In addition, we noted that spontaneous and induced BR control of lumbar sympathetic activity in the LP group was similar to the control group. The data indicate that hypertension in the adult rat offspring subjected to perinatal protein malnutrition is not related to baroreflex dysfunction.
    October 20, 2016   doi: 10.1111/1440-1681.12628   open full text
  • Exacerbated cardiac fibrosis induced by β‐adrenergic activation in old mice due to decreased AMPK activity.
    Jingjing Wang, Yao Song, Hao Li, Qiang Shen, Jing Shen, Xiangbo An, Jimin Wu, Jianshu Zhang, Yunong Wu, Han Xiao, Youyi Zhang.
    Clinical and Experimental Pharmacology and Physiology. October 20, 2016
    Senescent hearts exhibit defective responses to β‐adrenergic receptor (β‐AR) over‐activation upon stress, leading to more severe pathological cardiac remodelling. However, the underlying mechanisms remain unclear. Here, we investigated the role of adenosine monophosphate‐activated protein kinase (AMPK) in protecting against ageing‐associated cardiac remodelling in mice upon β‐AR over‐activation. 10‐week‐old (young) and 18‐month‐old (old) mice were subcutaneously injected with the β‐AR agonist isoproterenol (ISO; 5 mg/kg). More extensive cardiac fibrosis was found in old mice upon ISO exposure than in young mice. Meanwhile, ISO treatment decreased AMPK activity and increased β‐arrestin 1, but not β‐arrestin 2, expression, and the effects of ISO on AMPK and β‐arrestin 1 were greater in old mice than in young mice. Similarly, young AMPKα2‐knockout (KO) mice showed more extensive cardiac fibrosis upon ISO exposure than that was observed in age‐matched wild‐type (WT) littermates. The extent of cardiac fibrosis in WT old mice was similar to that in young KO mice. Additionally, AMPK activities were decreased and β‐arrestin 1 expression increased in KO mice. In contrast, the AMPK activator metformin decreased β‐arrestin 1 expression and attenuated cardiac fibrosis in both young and old mice upon ISO exposure. In conclusion, more severe cardiac fibrosis is induced by ISO in old mice than in young mice. A decrease in AMPK activity, which further increases β‐arrestin 1 expression, is the central mechanism underlying the ageing‐related cardiac fibrosis induced by ISO. The AMPK activator metformin is a promising therapeutic agent for treating ageing‐related cardiac remodelling upon β‐AR over‐activation.
    October 20, 2016   doi: 10.1111/1440-1681.12622   open full text
  • Role of mechanistic target of rapamycin (mTOR) in renal function and ischaemia–reperfusion induced kidney injury.
    Reem Alshaman, Luan Truong, Adebayo Oyekan.
    Clinical and Experimental Pharmacology and Physiology. October 20, 2016
    Despite the presence of many studies on the role of mechanistic target of rapamycin (mTOR) in cardiorenal tissues, the definitive role of mTOR in the pathogenesis of renal injury subsequent to ischaemia–reperfusion (IR) remains unclear. The aims of the current study were to characterize the role of mTOR in normal kidney function and to investigate the role of mTOR activation in IR‐induced kidney injury. In euvolemic anaesthetized rats, treatment with the mTOR inhibitor rapamycin increased blood pressure (121 ± 2 to 144 ± 3 mmHg; P<.05), decreased glomerular filtration rate (GFR; 1.6 ± 0.3 to 0.5 ± 0.2 mL/min; P<.05) and increased urinary sodium excretion (UNaV; 14 ± 1 to 109 ± 25 mmol/L per hour; P<.05). In rats subjected to IR, autophagy induction, p‐mTOR expression and serum creatinine increased (1.9 ± 0.2 to 3 ± 0.3 mg/dL; P<.05); treatment with rapamycin blunted p‐mTOR expression but further increased autophagy induction and serum creatinine (3 ± 0.3 to 5 ± 0.6 mg/dL; P<.05). In contrast, clenbuterol, an mTOR activator, blunted the effect of rapamycin on serum creatinine (4 ± 0.6 vs 2.3 ± 0.3 mg/dL; P<.05), autophagy induction and p‐mTOR expression. IR also increased 24 hour protein excretion (9 ± 3 to 17 ± 2 mg/day; P<.05) and kidney injury molecule‐1 (KIM‐1) expression, and rapamycin treatment further increased KIM‐1 expression. Clenbuterol exacerbated protein excretion (13 ± 2 to 26 ± 4 mg/day; P<.05) and antagonized the effect of rapamycin on KIM‐1 expression. Histopathological data demonstrated kidney injury in IR rats that was worsened by rapamycin treatment but attenuated by clenbuterol treatment. Thus, mTOR signalling is crucial for normal kidney function and protecting the kidney against IR injury through autophagy suppression.
    October 20, 2016   doi: 10.1111/1440-1681.12648   open full text
  • Chronic mercury exposure impairs the sympathovagal control of the rat heart.
    MR Simões, BF Azevedo, J Fiorim, DD Freire, EP Covre, DV Vassallo, L Santos.
    Clinical and Experimental Pharmacology and Physiology. October 20, 2016
    Mercury is known to cause harmful neural effects affecting the cardiovascular system. Here, we evaluated the chronic effects of low‐dose mercury exposure on the autonomic control of the cardiovascular system. Wistar rats were treated for 30 days with HgCl2 (1st dose 4.6 μg/kg followed by 0.07 μg/kg per day, intramuscular) or saline. The femoral artery and vein were then cannulated for evaluation of autonomic control of the hemodynamic function, which was evaluated in awake rats. The following tests were performed: baroreflex sensitivity, Von Bezold‐Jarisch reflex, heart rate variability (HRV) and pharmacological blockade with methylatropine and atenolol to test the autonomic tone of the heart. Exposure to HgCl2 for 30 days slightly increased the mean arterial pressure and heart rate (HR). There was a significant reduction in the baroreflex gain of animals exposed to HgCl2. Moreover, haemodynamic responses to the activation of the Von Bezold‐Jarisch reflex were also reduced. The changes in the spectral analysis of HRV suggested a shift in the sympathovagal balance toward a sympathetic predominance after mercury exposure, which was confirmed by autonomic pharmacological blockade in the HgCl2 group. This group also exhibited reduced intrinsic HR after the double block suggesting that the pacemaker activity of the sinus node was also affected. These findings suggested that the autonomic modulation of the heart was significantly altered by chronic mercury exposure, thus reinforcing that even at low concentrations such exposure might be associated with increased cardiovascular risk.
    October 20, 2016   doi: 10.1111/1440-1681.12624   open full text
  • Systemic inflammatory response syndrome following burns is mediated by brain natriuretic peptide/natriuretic peptide A receptor‐induced shock factor 1 signaling pathway.
    Yang‐Cheng Xu, Cheng‐Qun Luo, Xiong Li.
    Clinical and Experimental Pharmacology and Physiology. September 01, 2016
    The aim of this study was to determine whether systemic inflammatory response syndrome (SIRS) in burn patients is mediated by the brain natriuretic peptide (BNP)/natriuretic peptide A receptor (NPRA)‐induced heat shock factor 1 (HSF‐1) signalling pathway. Mononuclear cells (MNCs) that were isolated from patients with burn injuries and SIRS mouse models and a RAW264.7 cell line were treated with normal serum or serum obtained from animals with burn injuries. In parallel, small hairpin RNAs (shRNAs) against BNP or NPRA were transfected in both cell types. Western blotting (WB) and enzyme‐linked immunosorbent assay (ELISA) were used to detect protein expression and inflammatory factor levels, respectively. We found that interleukin (IL)‐12, tumour necrosis factor (TNF)‐α, C‐reactive protein (CRP), and BNP levels were increased and IL‐10 levels were decreased in the plasma and MNCs in vivo in the animal model of SIRS. Additionally, NPRA was upregulated, whereas HSF‐1 was downregulated in monocytes in vivo. Treatment of RAW264.7 cells with burn serum or BNP induced IL‐12, TNF‐α, and CRP secretion as well as HSF‐1 expression. Finally, silencing BNP with shRNA interrupted the effect of burn serum on RAW264.7 cells, and silencing NPRA blocked burn serum‐ and BNP‐mediated changes in RAW264.7 cells. These results suggest that the interaction of NPRA with BNP secreted from circulatory MNCs as well as mononuclear macrophages leads to inflammation via HSF‐1 during SIRS development following serious burn injury.
    September 01, 2016   doi: 10.1111/1440-1681.12620   open full text
  • Sex‐linked differences in the course of chronic kidney disease and congestive heart failure: a study in 5/6 nephrectomized Ren‐2 transgenic hypertensive rats with volume overload induced using aorto‐caval fistula.
    Luděk Červenka, Petra Škaroupková, Elzbieta Kompanowska‐Jezierska, Janusz Sadowski.
    Clinical and Experimental Pharmacology and Physiology. September 01, 2016
    The role of hypertension and the renin‐angiotensin system (RAS) in sex‐related differences in the course of chronic kidney disease (CKD) and congestive heart failure (CHF) remain unclear, especially when the two diseases are combined. In male and female Ren‐2 transgenic rats (TGR), a model of hypertension with activation of endogenous RAS, CKD was induced by 5/6 renal mass reduction (5/6 NX) and CHF was elicited by volume overload achieved by creation of an aorto‐caval fistula (ACF). The primary aim of the study was to examine long‐term CKD‐ and CHF‐related mortality, especially in animals with CKD and CHF combined, with particular interest in the potential sex‐related differences. The follow‐up period was 23 weeks after the first intervention (5/6 NX). We found, first, that TGR did not exhibit sexual dimorphism in the course of 5/6 NX‐induced CKD. Second, in contrast, TGR exhibited important sex‐related differences in the course of ACF‐induced CHF‐related mortality: intact female TGR showed higher survival rate than male TGR. This situation is reversed in the course of combined 5/6 NX‐induced CKD and ACF‐induced CHF‐related mortality: intact female TGR exhibited poorer survival than male TGR. Third, the survival rate in animals with combined 5/6 NX‐induced CKD and ACF‐induced CHF was significantly worsened as compared with rat groups that were exposed to ‘single organ disease’. Collectively, our present results clearly show that CKD aggravates long‐term mortality of animals with CHF. In addition, TGR exhibit remarkable sexual dimorphism with respect to CKD‐ and CHF‐related mortality, especially in animals with combined CKD and CHF.
    September 01, 2016   doi: 10.1111/1440-1681.12619   open full text
  • Renal cellular hypoxia in adenine‐induced chronic kidney disease.
    Debra Fong, Md Mahbub Ullah, Jaswini G. Lal, Amany Abdelkader, Connie P.C. Ow, Lucinda M. Hilliard, Sharon D. Ricardo, Darren J. Kelly, Roger G. Evans.
    Clinical and Experimental Pharmacology and Physiology. September 01, 2016
    We determined whether adenine‐induced chronic kidney disease (CKD) in rats is associated with renal tissue hypoxia. Adenine (100 mg) or its vehicle was administered to male Sprague‐Dawley rats, daily by oral gavage, over a 15‐day period. Renal function was assessed before, and 7 and 14 days after, adenine treatment commenced, by collection of a 24‐hour urine sample and a blood sample from the tail vein. On day 15, arterial pressure was measured in conscious rats via the tail artery. Renal tissue hypoxia was then assessed by pimonidazole adduct immunohistochemistry and fibrosis was assessed by staining tissue with picrosirius red and Masson's trichrome. CKD was evident within 7 days of commencing adenine treatment, as demonstrated by increased urinary albumin to creatinine ratio (30 ± 12‐fold). By day 14 of adenine treatment plasma creatinine concentration was more than 7‐fold greater, and plasma urea more than 5‐fold greater, than their baseline levels. On day 15, adenine‐treated rats had slightly elevated mean arterial pressure (8 mmHg), anaemia and renomegaly. Kidneys of adenine‐treated rats were characterised by the presence of tubular casts, dilated tubules, expansion of the interstitial space, accumulation of collagen, and tubulointerstitial hypoxia. Pimonidazole staining (hypoxia) co‐localised with fibrosis and was present in both patent and occluded tubules. We conclude that renal tissue hypoxia develops rapidly in adenine‐induced CKD. This model, therefore, should prove useful for examination of the temporal and spatial relationships between tubulointerstitial hypoxia and the development of CKD, and thus the testing of the ‘chronic hypoxia hypothesis’.
    September 01, 2016   doi: 10.1111/1440-1681.12621   open full text
  • Fixed dose of long‐acting erythropoietic stimulating agents at higher frequency improves appetite, reduces inflammation and corrects anaemia in patients on haemodialysis.
    Wen‐Sheng Liu, Da‐Chen Chu, Hsiang‐Lin Chan, Szu‐Yuan Li, Chih‐Kuang Liu, Chih‐Yu Yang, Yu‐Wei Chen, Pui‐Ching Lee, Yen‐Ting Lai, Chih‐Ching Lin.
    Clinical and Experimental Pharmacology and Physiology. September 01, 2016
    Anaemia is an important issue in patients undergoing haemodialysis. We aimed to identify a better dosing schedule of a fixed monthly dose of continuous erythropoietin receptor activator (CERA) in patients with chronic kidney disease (CKD) on haemodialysis. The CERA dosing schedule included 100 μg once monthly for 2 months, 50 μg twice monthly for 2 months and then 100 μg once monthly for two months. The effectiveness was determined by comparing haematocrit, nutritional status (serum protein and albumin) and inflammatory markers (tumour necrosis factor (TNF)‐α, interleukin (IL)‐1, IL‐6 and Hepcidin) at the beginning of the study with those at the end of the study. Forty‐seven out of 67 patients completed the trial. At the end, haematocrit was significantly higher (34.51 vs 33.22%, P=.004), levels of inflammatory markers were significantly lower (TNF‐α (30.71 vs 35.67 ng/mL, P=.007), IL‐6 (5.12 vs 7.95 ng/mL, P=.033), hepcidin (60.39 vs 74.39 ng/mL, P=.002)), blood glucose levels were significantly lower (112.40 vs 139.02 mg/dL, P=.003) and albumin was significantly higher (4.11 vs 3.98, P=.001). Patients with a better than average response had a lower initial number of red blood cells (3.3 vs 3.6 × 106/mm3, P=.025) and a lower IL‐1 (3.8 vs 12.9 ng/mL, P=.01). They also had significantly lower blood glucose levels at the end. (91.3 vs 124.0 mg/dL, P=.03). We demonstrate that a fixed monthly dose of CERA at a twice monthly dosing schedule improves nutrition, reduces the inflammation and corrects anaemia in patients on haemodialysis. This finding may provide a new strategy for treating CKD‐related anaemia.
    September 01, 2016   doi: 10.1111/1440-1681.12618   open full text
  • Protein tyrosine phosphatase PTPRB regulates Src phosphorylation and tumour progression in NSCLC.
    Yinliang Qi, Yuanchang Dai, Shuyu Gui.
    Clinical and Experimental Pharmacology and Physiology. September 01, 2016
    Protein tyrosine‐phosphatases (PTPs) play important roles in various biological processes. Deregulation in PTP function has been implicated in carcinogenesis and tumour progression in many cancer types. However, the role of protein tyrosine phosphatase receptor type B (PTPRB) in non‐small‐cell lung cancer (NSCLC) tumorigenesis has not been investigated. Lentiviral vector expressing PTPRB cDNA or shRNA was infected into A549 and H1299 cell lines, followed by cell proliferation, colony formation, soft agar and invasion assays. A549 xenograft mouse model was used to evaluate in vivo function of PTPRB. Quantitative polymerase chain reaction (PCR) was used to measure PTPRB expression in NSCLC patient samples. Kaplan Meier analysis was performed to assess association between PTPRB expression and patient overall survival (OS). Multivariate analysis was performed to evaluate prognostic significance of PTPRB. Overexpression of PTPRB reduced cell proliferation rate, colony formation efficiency, soft agar growth and cell invasion in A549 and H1299 cells, as well as tumour growth rate in A549 xenograft. Knockdown of PTPRB increased Src phosphorylation and cell invasion, which was reversed by Src inhibitor PP2. Additionally, PTPRB was down‐regulated in NSCLC patient and was associated with patient OS. PTPRB regulates Src phosphorylation and tumorigenesis in NSCLC. PTPRB may serve as an independent prognostic biomarker for NSCLC patients.
    September 01, 2016   doi: 10.1111/1440-1681.12610   open full text
  • MicroRNA‐217 suppresses homocysteine‐induced proliferation and migration of vascular smooth muscle cells via N‐methyl‐D‐aspartic acid receptor inhibition.
    Hongyan Duan, Yongqiang Li, Lijie Yan, Haitao Yang, Jintao Wu, Peng Qian, Bing Li, Shanling Wang.
    Clinical and Experimental Pharmacology and Physiology. September 01, 2016
    Hyperhomocysteine has become a critical risk for atherosclerosis and can stimulate proliferation and migration of vascular smooth muscle cells (VSMCs). N‐methyl‐D‐aspartic acid receptor (NMDAR) is a receptor of homocysteine and mediates the effects of homocysteine on VSMCs. Bioinformatics analysis has shown NMDAR is a potential target of microRNA‐217 (miR‐217), which exerts multiple functions in cancer tumorigenesis and carotid plaque progression. In this study, we sought to investigate the role of miR‐217 in VSMCs phenotype transition under homocysteine exposure and elucidate its effect on atherosclerotic plaque formation. After treating with several doses of homocysteine (0–8 × 10−4 mol/L) for 24 hours, the expression of miR‐217 in HA‐VSMCs and rat aortic VSMCs was not altered. Intriguingly, the expression of NMDAR mRNA and protein was reduced by homocysteine in a dose‐dependent manner. Transfection of miR‐217 mimic significantly inhibited the proliferation and migration of VSMCs with homocysteine treatment, while transfection of miR‐217 inhibitor promoted VSMCs migration. Moreover, miR‐217 mimic down‐regulated while miR‐217 inhibitor up‐regulated NMDAR protein expression but not NMDAR mRNA expression. Through luciferase reporter assay, we showed that miR‐217 could directly bind to the 3′‐UTR of NMDAR. MiR‐217 mimic transfection also released the inhibition of cAMP‐response element‐binding protein (CREB)‐PGC‐1α signalling induced by homocysteine. Additionally, restoration of PGC‐1α expression via AdPGC‐1α infection markedly suppressed VSMCs proliferation through the degradation of NADPH oxidase (NOX1) and reduction of reactive oxygen species (ROS). Collectively, our study identified the role of miR‐217 in regulating VSMCs proliferation and migration, which might serve as a target for atherosclerosis therapy.
    September 01, 2016   doi: 10.1111/1440-1681.12611   open full text
  • Iron‐chelating agent, deferasirox, inhibits neutrophil activation and extracellular trap formation.
    Mari Kono, Katsuyasu Saigo, Shiori Yamamoto, Kohei Shirai, Shuta Iwamoto, Tomoko Uematsu, Takayuki Takahashi, Shion Imoto, Makoto Hashimoto, Yosuke Minami, Atsushi Wada, Mariko Takenokuchi, Seiji Kawano.
    Clinical and Experimental Pharmacology and Physiology. September 01, 2016
    Iron‐chelating agents, which are frequently prescribed to transfusion‐dependent patients, have various useful biological effects in addition to chelation. Reactive oxygen species (ROS) produced by neutrophils can cause pulmonary endothelial cell damage, which can lead to acute lung injury (ALI). We previously reported that deferasirox (DFS), an iron‐chelating agent, inhibits phorbol myristate acetate (PMA) or formyl‐methionyl‐leucyl‐phenylalanine (fMLP)‐induced ROS production in neutrophils, in vitro. Here, we investigate whether DFS inhibits vacuolization in neutrophils and neutrophil extracellular trap (NET) formation. Human neutrophils were incubated with DFS and stimulated with PMA or fMLP. Human neutrophils were separated from heparinized peripheral blood using density gradient centrifugation, and subsequently incubated with DFS. After 10 minutes, neutrophils were stimulated by PMA or fMLP. Vacuole formation was observed by electron microscopy. For observing NET formations using microscopes, immunohistological analyses using citrullinated histone H3 and myeloperoxidase antibodies, and SYTOX Green (an impermeable DNA detection dye) staining, were conducted. NET formation was measured as the quantity of double‐stranded DNA (dsDNA), using the AccuBlue Broad Range dsDNA Quantitation Kit. DFS (50 μmol/L) inhibited vacuole formation in the cytoplasm and NET formation. Additionally, 5–100 μmol/L concentration of DFS inhibited the release of dsDNA in a dose‐independent manner. We demonstrate that DFS inhibits not only ROS production but also vacuolization and NET formation in neutrophils. These results suggest the possibility of protective effects of DFS against NET‐related adverse effects, including ALI and thrombosis.
    September 01, 2016   doi: 10.1111/1440-1681.12612   open full text
  • A novel translocator protein 18 kDa ligand, ZBD‐2, exerts neuroprotective effects against acute spinal cord injury.
    Qiang Cheng, Guo‐jing Sun, Shui‐bing Liu, Qi Yang, Xiao‐ming Li, Xu‐bo Li, Gang Liu, Jian‐ning Zhao, Ming‐gao Zhao.
    Clinical and Experimental Pharmacology and Physiology. September 01, 2016
    Traumatic spinal cord injury (SCI) happens accidently and often leads to motor dysfunction due to a series of biochemical and pathological events and damage, either temporarily or permanently. Translocator protein 18 (TSPO) has been found to be involved in the synthesis of endogenous neurosteroids which have multiple effects on neurons, but the internal mechanisms are not clear. N‐benzyl‐N‐ethyl‐2‐(7,8‐oxo‐2‐phenyl‐9H‐purin‐9‐yl) acetamide (ZBD‐2), a newly reported ligand of TSPO, shows some neuroprotective effect against focal cerebral ischemia in vivo and NMDA‐induced neurotoxicity in vitro. The present study aims to examine the role of ZBD‐2 in SCI mice and elucidate the underlying molecular mechanisms. The SCI model was established by crushing spinal cord. ZBD‐2 (10 mg/kg) significantly enhanced the hindlimb locomotor functions after SCI and decreased the tissue damage and conserved the white matter of the spinal cord. High‐dose ZBD‐2 alleviated the oxidative stress induced by SCI and regulated the imbalance between NR2B‐containing NMDA and GABA receptors by increasing the levels of GAD67 in the spinal cord of SCI mice. Additionally, ZBD‐2 (10 mg/kg) increased phosphorylated Akt (p‐Akt) and decreased the ratio of Bax/Bcl‐2. These results demonstrate that ZBD‐2 performs neuroprotection against SCI through regulating the synaptic transmission and the PI3K/AKT signaling pathway.
    September 01, 2016   doi: 10.1111/1440-1681.12606   open full text
  • VCP746, a novel A1 adenosine receptor biased agonist, reduces hypertrophy in a rat neonatal cardiac myocyte model.
    Chung H Chuo, Shane M Devine, Peter J Scammells, Henry Krum, Arthur Christopoulos, Lauren T May, Paul J White, Bing H Wang.
    Clinical and Experimental Pharmacology and Physiology. September 01, 2016
    VCP746 is a novel A1 adenosine receptor (A1AR) biased agonist previously shown to be cytoprotective with no effect on heart rate. The aim of this study was to investigate the potential anti‐hypertrophic effect of VCP746 in neonatal rat cardiac myocytes (NCM). NCM hypertrophy was stimulated with interleukin (IL)‐1β (10 ng/mL), tumour necrosis factor (TNF)‐α (10 ng/mL) or Ang II (100 nmol/L) and was assessed by 3H‐leucine incorporation assay. VCP746 significantly inhibited IL‐1β‐, TNF‐α‐ and Ang II‐stimulated NCM hypertrophy as determined by 3H‐leucine incorporation. The anti‐hypertrophic effect of VCP746 was also more potent than that of the prototypical A1AR agonist, N6‐cyclopentyladenosine (CPA). Further investigation with the 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT) cell viability assay showed that neither CPA nor VCP746 had any effect on cell viability, confirming that the reduction in 3H‐leucine incorporation mediated by CPA and VCP746 was not due to a reduction in cell viability. IL‐1β, TNF‐α and Ang II were also shown to increase the mRNA expression of hypertrophy biomarkers, ANP, β‐MHC and α‐SKA in NCM. Treatment with VCP746 at concentrations as low as 1 nmol/L suppressed mRNA expression of ANP, β‐MHC and α‐SKA stimulated by IL‐1β, TNF‐α or Ang II, demonstrating the broad mechanistic basis of the potent anti‐hypertrophic effect of VCP746. This study has shown that the novel A1AR agonist, VCP746, is able to attenuate cardiac myocyte hypertrophy. As such, VCP746 is potentially useful as a pharmacological agent in attenuating cardiac remodelling, especially in the post‐myocardial infarction setting, given its previously established cytoprotective properties.
    September 01, 2016   doi: 10.1111/1440-1681.12616   open full text
  • The role of apelin in central cardiovascular regulation in rats with post‐infarct heart failure maintained on a normal fat or high fat diet.
    Katarzyna Czarzasta, Agnieszka Cudnoch‐Jedrzejewska, Ewa Szczepanska‐Sadowska, Lukasz Fus, Liana Puchalska, Agata Gondek, Jakub Dobruch, Ryszard Gomolka, Robert Wrzesien, Tymoteusz Zera, Barbara Gornicka, Marek Kuch.
    Clinical and Experimental Pharmacology and Physiology. September 01, 2016
    Based on the available literature, it can be assumed that in cases of post‐infarct heart failure (HF) and obesity, a significant change in the central regulation of the cardiovascular system takes place with, among others, the involvement of the apelinergic system. The main objective of the present study was to clarify the role of apelin‐13 in the central regulation of the cardiovascular system in Sprague Dawley rats with HF or sham operated (SO) and fed on a normal fat (NFD) or a high fat diet (HFD). The study was divided into two parts: Part I, hemodynamic studies; and Part II, biochemical and molecular studies. The animals were subjected to the following research procedures. Part I and II: feeding NFD or HFD; experimental induction of HF or SO; Part I: intracerebroventricular (ICV) infusion of the examined substances, monitoring of mean arterial blood pressure (MABP) and heart rate (HR); Part II: venous blood and tissue samples collected. ICV infusion of apelin‐13 caused significantly higher changes in ΔMABP in the SO NFD group. No changes were noted in ΔHR in any of the studied groups. Apelin and apelin receptor (APJ) mRNA expression in the brain and adipose tissues was higher in the HF rats. HFD causes significant increase in expression of apelin and APJ mRNA in the left ventricle. In conclusion, HF and HFD appear to play an important role in modifying the activity of the central apelinergic system and significant changes in mRNA expression of apelin and APJ receptor.
    September 01, 2016   doi: 10.1111/1440-1681.12617   open full text
  • Human mesenchymal stem cells attenuate pulmonary hypertension induced by prenatal lipopolysaccharide treatment in rats.
    Hsiu‐Chu Chou, Willie Lin, Chung‐Ming Chen.
    Clinical and Experimental Pharmacology and Physiology. September 01, 2016
    Intra‐amniotic injection of lipopolysaccharide (LPS) induces pulmonary hypertension in newborn rats. This study was designed to test whether human mesenchymal stem cells (MSCs) reduce pulmonary hypertension and alleviate cardiac hypertrophy in prenatal LPS‐treated rats. Pregnant Sprague‐Dawley rats were injected intraperitoneally with LPS (0.5 mg/kg per day) or untreated on gestational days 20 and 21. Human MSCs (3×105 cells and 1×106 cells) in 0.03 mL of normal saline (NS) were transplanted intratracheally on postnatal day 5. Four study groups were considered: normal, LPS+NS, LPS+MSCs (3×105 cells), and LPS+MSCs (1×106 cells). On postnatal day 14, lung and heart tissues were collected for measuring the arterial medial wall thickness (MWT) and β‐myosin heavy chain (β‐MHC) level as markers of pulmonary hypertension and cardiac hypertrophy, respectively. The LPS+NS group exhibited a significantly higher right ventricle (RV)/[left ventricle (LV)+ interventricular septum (IVS)] thickness ratio and MWT, a greater cardiomyocyte width, a greater number of cardiomyocyte nuclei per squared millimeter, and higher β‐MHC expression than those observed in the normal group. Human MSC transplantation (3×105 cells and 1×106 cells) in LPS‐treated rats reduced MWT and the RV/(LV+IVS) thickness ratio to normal levels. This improvement in right ventricular hypertrophy was accompanied by a decrease in toll‐like receptor 4 (TLR4), nuclear factor‐κB, and tumor necrosis factor‐α expression in the heart. Intratracheal human MSCs transplantation can attenuate pulmonary hypertension and right ventricular hypertrophy in prenatal LPS‐treated rats; this attenuation may be associated with suppression of TLR4 expression via paracrine pathways.
    September 01, 2016   doi: 10.1111/1440-1681.12604   open full text
  • Histone deacetylase inhibition, but not a mineralocorticoid receptor antagonist spironolactone, attenuates atypical transcription by an activating mutant MR (MRS810L).
    Seol‐Hee Kang, Hae‐Ahm Lee, Eunjo Lee, Mina Kim, Inkyeom Kim.
    Clinical and Experimental Pharmacology and Physiology. September 01, 2016
    A mutation in the mineralocorticoid receptor (MRS810L) leads to early‐onset hypertension, which is markedly exacerbated during pregnancy. The mutation causes progesterone and even the MR antagonist spironolactone to become potent agonists. Thus, it is hard to control hypertension in patients harbouring this mutation. We hypothesized that histone deacetylase inhibition (HDACi), but not the MR antagonist spironolactone, attenuates atypical transcriptional activity of activating mutant MR (MRS810L). We established HEK293T cells overexpressing wild‐type MR (MRWT) or MRS810L and determined their transcriptional activities by luciferase assay. Expression of MR target genes was measured by quantitative real‐time PCR (qRT‐PCR). Treatment with aldosterone increased the expression of MR target genes as well as the transcriptional activities in HEK293T cells transfected either with MRWT or MRS810L. Treatment with either spironolactone or progesterone also increased the expression of MR target genes as well as transcriptional activity, but only in HEK293T cells transfected with MRS810L. Spironolactone abolished the promoter activity stimulated by aldosterone in HEK293T cells transfected with MRWT. Treatment with HDAC inhibitors attenuated the transcriptional activity as well as the expression of MR target genes induced by aldosterone, spironolactone, or progesterone whether HEK293T cells were transfected with either MRWT or MRS810L. These results indicate that HDACi, but not an MR antagonist spironolactone, attenuates atypical transcriptional activity of an activating mutant MR (MRS810L).
    September 01, 2016   doi: 10.1111/1440-1681.12614   open full text
  • Glutamate protects against Ca2+ paradox‐induced injury and inhibits calpain activity in isolated rat hearts.
    Jian‐Ying Zhang, Ling‐Heng Kong, Dong Lai, Zhen‐Xiao Jin, Xiao‐Ming Gu, Jing‐Jun Zhou.
    Clinical and Experimental Pharmacology and Physiology. September 01, 2016
    This study determined the effects of glutamate on the Ca2+ paradoxical heart, which is a model for Ca2+ overload‐induced injury during myocardial ischaemia and reperfusion, and evaluated its effect on a known mediator of injury, calpain. An isolated rat heart was retrogradely perfused in a Langendorff apparatus. Ca2+ paradox was elicited via perfusion with a Ca2+‐free Krebs‐Henseleit (KH) solution for 3 minutes followed by Ca2+‐containing normal KH solution for 30 minutes. The Ca2+ paradoxical heart exhibited almost no viable tissue on triphenyltetrazolium chloride staining and markedly increased LDH release, caspase‐3 activity, cytosolic cytochrome c content, and apoptotic index. These hearts also displayed significantly increased LVEDP and a disappearance of LVDP. Glutamate (5 and 20 mmol/L) significantly alleviated Ca2+ paradox‐induced injury. In contrast, 20 mmol/L mannitol had no effect on Ca2+ paradox. Ca2+ paradox significantly increased the extent of the translocation of μ‐calpain to the sarcolemmal membrane and the proteolysis of α‐fodrin, which suggests calpain activation. Glutamate also blocked these effects. A non‐selective inhibitor of glutamate transporters, dl‐TBOA (10 μmol/L), had no effect on control hearts, but it reversed glutamate‐induced cardioprotection and reduction in calpain activity. Glutamate treatment significantly increased intracellular glutamate content in the Ca2+ paradoxical heart, which was also blocked by dl‐TBOA. We conclude that glutamate protects the heart against Ca2+ overload‐induced injury via glutamate transporters, and the inhibition of calpain activity is involved in this process.
    September 01, 2016   doi: 10.1111/1440-1681.12605   open full text
  • Daphnetin inhibits TNF‐α and VEGF‐induced angiogenesis through inhibition of the IKKs/IκBα/NF‐κB, Src/FAK/ERK1/2 and Akt signalling pathways.
    Abhishek Kumar, Priyashree Sunita, Shivesh Jha, Shakti Prasad Pattanayak.
    Clinical and Experimental Pharmacology and Physiology. September 01, 2016
    Coumarins, identified as plant secondary metabolites possess diverse biological activities including anti‐angiogenic properties. Daphnetin (DAP), a plant derived dihydroxylated derivative of coumarin has shown significant pharmacological properties such as anticancer, anti‐arthritic and anti‐inflammatory. The present study was performed to investigate the anti‐angiogenic potential of DAP, focusing on the mechanism of action. The in vivo anti‐angiogenic potential of DAP was evaluated by vascular endothelial growth factor (VEGF)‐induced rat aortic ring (RAR) assay and chick chorioallantoic membrane (CAM) assay. For in vitro evaluation, wounding migration, transwell invasion, tube formation and apoptosis assays were performed on VEGF (8 ng/mL)‐induced human umbilical vein endothelial cells (HUVECs). The cellular mechanism of DAP was examined on TNFα (10 ng/mL) and VEGF‐induced HUVECs by extracting the mRNA and protein levels using RT‐qPCR and western blotting. Our data demonstrated that DAP inhibited the in vivo angiogenesis in the RAR and CAM assay. DAP also inhibited the different steps of angiogenesis, such as migration, invasion, and tube formation in HUVECs. DAP inhibited nuclear factor‐κB signalling together including TNF‐α induced IκBα degradation; phosphorylation of IκB kinase (IKKα/β) and translocation of the NF‐κB‐p65 protein. Furthermore, western blotting revealed that DAP significantly down‐regulated the VEGF‐induced signalling such as c‐Src, FAK, ERK1/2 and the related phosphorylation of protein kinase B (Akt) and VEGFR2 expressions. DAP reduced the elevated mRNA expression of iNOS, MMP2 and also, induced apoptosis in VEGF‐stimulated HUVECs by the caspase‐3 dependent pathway. Taken together, this study reveals that DAP may have novel prospective as a new multi‐targeted medication for the anti‐angiogenesis and cancer therapy.
    September 01, 2016   doi: 10.1111/1440-1681.12608   open full text
  • Overexpression of copper/zinc superoxide dismutase in the median preoptic nucleus improves cardiac function after myocardial infarction in the rat.
    John P Collister, Cristina Hartnett, Tim Mayerhofer, David Nahey, Christopher Stauthammer, Maxie Krüger, Anthony Tobias, M. Gerard O'Sullivan, Josh Parker, Jun Tian, Adam J Case, Matthew C Zimmerman.
    Clinical and Experimental Pharmacology and Physiology. September 01, 2016
    Previous reports indicate that overexpression of copper/zinc superoxide dismutase (CuZnSOD), an intracellular superoxide (O2•−) scavenging enzyme, in the brain subfornical organ improves cardiac function in a mouse model of heart failure (HF). A downstream hypothalamic site, the MnPO, may act as a relay centre for O2•− to serve as a mediator in the pathophysiology of HF. To test the hypothesis that elevated O2•− in the MnPO contributes to the pathophysiology of HF and decreased cardiac function, we injected adenovirus encoding CuZnSOD (AdCuZnSOD, n=7) or control empty adenovirus vector (AdEmpty, n=7) into the MnPO of normal rats. Subsequently, rats were subjected to coronary artery ligation to create a myocardial infarct (MI) of the left ventricle. Cardiac function was monitored via echocardiography. Upon completion, rat brains were examined for CuZnSOD expression in MnPO via immunofluorescence and histopathological analyses of cardiac infarct size were conducted. Baseline (EF) ejection fractions (%) of AdCuZnSOD and AdEmpty rats were 73 ± 1 and 71 ± 1, respectively. Two weeks after MI, EF was significantly decreased in both groups of rats (AdCuZnSOD: 51 ± 3, AdEmpty: 46 ± 1). In contrast, by 4 weeks post MI, EF had improved to 64 ± 2 in AdCuZnSOD rats, yet was only 52 ± 1 in AdEmpty rats, and this was accompanied by lower plasma noradrenaline levels in AdCuZnSOD rats (0.49 ± 0.19 ng/mL) compared to AdEmpty rats (1.20 ± 0.32 ng/mL). In conclusion, despite decreases in EF early after MI, overexpression of CuZnSOD in the MnPO was related to an improvement in left ventricular function and concomitant decreased plasma noradrenaline levels 4 weeks post MI.
    September 01, 2016   doi: 10.1111/1440-1681.12607   open full text
  • Renal simplicity denervation reduces blood pressure and renal injuries in an obesity‐induced hypertension dog model.
    Zhihui Zhang, Kan Yang, Lixiong Zeng, Xiaoyan Wang, Fenglin Jiang, Shan Tu, Qishun Liang, Zhijie Shen.
    Clinical and Experimental Pharmacology and Physiology. August 25, 2016
    This study aimed to investigate the effects of renal sympathetic denervation (RDN) on blood pressure, renal function, and renal tissue pathological changes in obesity‐induced hypertensive dogs. Thirty‐two beagle dogs (10‐12 months) were randomized to the control (n=10) and model groups (n=22). High‐fat diet (HFD) was used to establish the obesity‐induced hypertensive model. After 3 months of HFD, 20 animals with successfully induced hypertension were randomized to the RDN (n=10) and sham groups (n=10). Renal artery angiography, body weight, blood pressure, heart rate, and blood and urine biochemistry were determined 1, 3, and 6 months after surgery. Models were sacrificed 6 months after surgery. Pathological changes in the renal artery and renal tissue were assessed. The HFD group had significantly (P<0.05) increased body weight, heart rate, and blood pressure, and higher levels of urine albumin, serum norepinephrine, and angiotensin II compared with controls. After RDN, blood pressure was decreased compared with baseline and the sham group (P<0.05). In the RDN group, examination of the renal artery and renal tissue showed intact intima of renal artery in the surgical area, renal sympathetic nerve degeneration, necrosis, and dissolution, and widened space between nerve fibers. Hypertension‐induced renal pathological changes were mild to moderate in the RDN group, but severe in the sham group. The control group had normal glomerular structure. In conclusion, RDN can effectively lower blood pressure in obesity‐induced hypertensive dogs, as well as hypertension‐induced renal pathological changes. This article is protected by copyright. All rights reserved.
    August 25, 2016   doi: 10.1111/1440-1681.12661   open full text
  • Hyperoxia‐mediated LC3B activation contributes to the impaired transdifferentiation of type II alveolar epithelial cells (AECIIs) to type I cells (AECIs).
    Liang Zhang, Shuang Zhao, Lijie Yuan, Hongmin Wu, Hong Jiang, Gang Luo.
    Clinical and Experimental Pharmacology and Physiology. July 26, 2016
    Life‐saving mechanical ventilation can also cause lung injury through the overproduction of reactive oxygen species (ROS), leading to bronchopulmonary dysplasia (BPD)‐like symptoms in preterm infants. It is reported that the autophagic protein microtubule‐associated protein‐1 light chain (LC)‐3B can confer protection against hyperoxia‐induced DNA damage in lung alveolar epithelium. However, its role in the transdifferentiation of type II alveolar epithelial cells (AECIIs) to type I cells (AECIs) is unclear and requires further investigation. In this study, newborn Sprague‐Dawley rats were exposed to 90% oxygen for up to 14 days to mimic BPD in human infants, with neonatal pups exposed to room air (21% oxygen) as controls. Primary rat AECIIs were cultured under hyperoxic conditions for up to 24 hours to further investigate the underlying mechanisms. This study found that hyperoxia promoted a significant and time‐dependent increase of AECII marker surfactant protein (SP)‐C in the lung. The increase of AECI marker T1α was repressed by hyperoxia during lung development. These results indicated an impaired AECII transdifferentiation. Pulmonary ROS concentration and expression of autophagic protein LC‐3B were increased gradually in response to hyperoxia exposure. Furthermore, AECIIs produced more ROS when cultured under hyperoxic conditions in vitro. Both the LC3B expression and the conversion from LC3BI to LC3BII were enhanced in hyperoxic AECs. Interestingly, inhibition of LC3B either by ROS inhibitor N‐acetyl‐l‐cysteine (NAC) or adenovirus‐mediated LC3B shRNA could partly restore AECII transdifferentiation under hyperoxia condition. In summary, the current study reveals a novel role of activated LC3B induced by hyperoxia in AECII transdifferentiation.
    July 26, 2016   doi: 10.1111/1440-1681.12592   open full text
  • Pharmacological rescue of hERG currents carried out by G604S and wide type hERG co‐expression.
    Jianhua Huo, Aifeng Zhang, Xueyan Guo, Hua Qiang, Ping Liu, Ling Bai, Aiqun Ma.
    Clinical and Experimental Pharmacology and Physiology. July 26, 2016
    Mutations in human ether‐a‐go‐go‐related gene (hERG) can lead to type 2 long‐QT syndrome (LQT2). The authors previously identified the hERG mutation G604S results in a loss of function and obviously decreased current amplitude and impaired channel protein trafficking when co‐expressed with WT‐hERG. The present study further investigates the biological and electrophysiological consequences of pharmacologic chaperones in HEK293 cells expressing G604S‐hERG or co‐expressing G604S‐hERG and WT‐hERG. It was found that a low temperature (27°C), thapsigargin, NS1643 and E‐4031 fail to rescue the G604S mutation. Interestingly, only E‐4031 treatment resulted in a significant increase in hERG currents in cells co‐expressing G604S‐hERG and WT‐hERG, correspondingly more mature protein band at 155 kDa by Western blotting and an increased membrane staining by confocal microscopy. In addition, E‐4031 treatment shifted the steady‐state half maximal activation voltage (V1/2) of the inactivation curve by +8 mV in cells co‐expressing G604S‐hERG and WT‐hERG. The present experimental results suggest that a G604S mutation is resistant to pharmacological rescue. E‐4031 treatment resulted in a significant increase in hERG currents by promoting the hERG channel processing and trafficking in cells co‐expressing G604S‐hERG and WT‐hERG.
    July 26, 2016   doi: 10.1111/1440-1681.12593   open full text
  • Orchidectomy attenuates high‐salt diet‐induced increases in blood pressure, renovascular resistance, and hind limb vascular dysfunction: role of testosterone.
    Ahmed K Oloyo, Olusoga A Sofola, Momoh A Yakubu.
    Clinical and Experimental Pharmacology and Physiology. July 26, 2016
    Sex hormone‐dependent vascular reactivity is an underlying factor contributing to sex differences in salt‐dependent hypertension. This study evaluated the role of androgens (testosterone) in high salt‐induced increase in blood pressure (BP) and altered vascular reactivity in renal blood flow and perfused hind limb preparation. Weanling male rats (8 weeks old, 180–200 g) were bilaterally orchidectomised or sham operated with or without testosterone replacement (Sustanon 250, 10 mg/kg intramuscularly once in 3 weeks) and placed on a normal (0.3%) or high (4.0%) NaCl diet for 6 weeks. The high‐salt diet (HSD) increased arterial BP, renal vascular resistance (RVR) and positive fluid balance (FB). These changes were accompanied by decreased plasma nitric oxide levels. The increased BP, RVR and FB observed in the rats fed a HSD were reversed by orchidectomy while testosterone replacement prevented the reversal. Phenylephrine (PE)‐induced increased vascular resistance in the perfused hind limb vascular bed was enhanced by HSD, the enhanced vascular resistance was prevented by orchidectomy and testosterone replacement reversed orchidectomy effect. Vasorelaxation responses to acetylcholine (ACh) and sodium nitroprusside (SNP) were impaired in HSD groups, orchidectomy attenuated the impairment, while testosterone replacement prevented the orchidectomy attenuation. These data suggested that eNOS‐dependent and independently‐mediated pathways were equally affected by HSD in vascular function impairment and this effect is testosterone‐dependent in male Sprague‐Dawley rats.
    July 26, 2016   doi: 10.1111/1440-1681.12595   open full text
  • The role of vagal pathway and NK1 and NK2 receptors in cardiovascular and respiratory effects of neurokinin A.
    Katarzyna Kaczyńska, Monika Jampolska, Małgorzata Szereda‐Przestaszewska.
    Clinical and Experimental Pharmacology and Physiology. July 26, 2016
    Neurokinin A (NKA) is a peptide neurotransmitter that participates in the regulation of breathing and the cardiovascular system. The purpose of the current study was to determine the cardiorespiratory pattern exerted by the systemic injection of NKA, to look at the contribution of neurokinin NK1 and NK2 receptors, and to establish the engagement of the vagal pathway in mediation of these responses. The effects of intravenous injections of NKA (50 μg/kg) were studied in anaesthetized, spontaneously breathing rats in the following experimental schemes: in neurally intact rats; and vagotomized at either midcervical or supranodosal level. Intravenous injections of NKA in the intact rats evoked sudden and short‐lived increase in the respiratory rate concomitant with drop in tidal volume, followed by a prolonged depression, coupled with continuous augmentation of the tidal volume. Respiratory alterations were accompanied by transient tachycardia and prolonged hypotension. Midcervical vagotomy eliminated respiratory rate response and augmentation of tidal volume. Section of supranodosal vagi abrogated all respiratory reactions. NK2 receptor blockade abolished respiratory changes without affecting cardiovascular effects, whereas NK1 receptor blockade significantly reduced hypotension and increase in heart rate with no impact on the respiratory system. These results indicate that NKA induced changes in the breathing resulting from an excitation of the NK2 receptors on the vagal endings. A fall in blood pressure triggered by NKA occurs outside of the vagus nerve and is probably mediated via its direct action on vascular smooth muscles supplied with NK1 receptors.
    July 26, 2016   doi: 10.1111/1440-1681.12594   open full text
  • Effects of transthyretin on thyroxine and β‐amyloid removal from cerebrospinal fluid in mice.
    Ruoli Chen, Carl P Chen, Jane E Preston.
    Clinical and Experimental Pharmacology and Physiology. July 26, 2016
    Transthyretin (TTR) is a binding protein for the thyroid hormone thyroxine (T4), retinol and β‐amyloid peptide. TTR aids the transfer of T4 from the blood to the cerebrospinal fluid (CSF), but also prevents T4 loss from the blood‐CSF barrier. It is, however, unclear whether TTR affects the clearance of β‐amyloid from the CSF. This study aimed to investigate roles of TTR in β‐amyloid and T4 efflux from the CSF. Eight‐week‐old 129sv male mice were anaesthetized and their lateral ventricles were cannulated. Mice were infused with artificial CSF containing 125I‐T4/3H‐mannitol, or 125I‐Aβ40/3H‐inulin, in the presence or absence of TTR. Mice were decapitated at 2, 4, 8, 16, 24 minutes after injection. The whole brain was then removed and divided into different regions. The radioactivities in the brain were determined by liquid scintillation counting. At baseline, the net uptake of 125I‐T4 into the brain was significantly higher than that of 125I‐Aβ40, and the half time for efflux was shorter (125I‐T4, 5.16; 3H‐mannitol, 7.44; 125I‐Aβ40, 8.34; 3H‐inulin, 10.78 minutes). The presence of TTR increased the half time for efflux of 125I‐T4 efflux, and caused a noticeable increase in the uptake of 125I‐T4 and 125I‐Aβ40 in the choroid plexus, whilst uptakes of 3H‐mannitol and 3H‐inulin remained similar to control experiments. This study indicates that thyroxine and amyloid peptide effuse from the CSF using different transporters. TTR binds to thyroxine and amyloid peptide to prevent the loss of thyroxine from the brain and redistribute amyloid peptide to the choroid plexus.
    July 26, 2016   doi: 10.1111/1440-1681.12598   open full text
  • Alterations of voltage‐dependent K+ channels in the mesenteric artery during the early and chronic phases of diabetes.
    Da Hye Hong, Hongliang Li, Hye Won Kim, Han Sol Kim, Youn Kyoung Son, Se‐Ran Yang, Jeong‐Ran Park, Kwon‐Soo Ha, Eun‐Taek Han, Seok‐Ho Hong, Amy L. Firth, Sung Hun Na, Won Sun Park.
    Clinical and Experimental Pharmacology and Physiology. July 26, 2016
    This study investigated the alteration of voltage‐dependent K+ (Kv) channels in mesenteric arterial smooth muscle cells from control (Long‐Evans Tokushima Otsuka [LETO]) and diabetic (Otsuka Long‐Evans Tokushima Fatty [OLETF]) rats during the early and chronic phases of diabetes. We demonstrated alterations in the mesenteric Kv channels during the early and chronic phase of diabetes using the patch‐clamp technique, the arterial tone measurement system, and RT‐PCR in Long‐Evans Tokushima (LETO; for control) and Otsuka Long‐Evans Tokushima Fatty (OLETF; for diabetes) type 2 diabetic model rats. In the early phase of diabetes, the amplitude of mesenteric Kv currents induced by depolarizing pulses was greater in OLETF rats than in LETO rats. The contractile response of the mesenteric artery induced by the Kv inhibitor, 4‐aminopyridine (4‐AP), was also greater in OLETF rats. The expression of most Kv subtypes‐ including Kv1.1, Kv1.2, Kv1.4, Kv1.5, Kv1.6, Kv2.1, Kv3.2, Kv4.1, Kv4.3, Kv5.1, Kv6.2, Kv8.1, Kv9.3, and Kv10.1‐were increased in mesenteric arterial smooth muscle from OLETF rats compared with LETO rats. However, in the chronic phase of diabetes, the Kv current amplitude did not differ between LETO and OLETF rats. In addition, the 4‐AP‐induced contractile response of the mesenteric artery and the expression of Kv subtypes did not differ between the two groups. The increased Kv current amplitude and Kv channel‐related contractile response were attributable to the increase in Kv channel expression during the early phase of diabetes. The increased Kv current amplitude and Kv channel‐related contractile response were reversed during the chronic phase of diabetes.
    July 26, 2016   doi: 10.1111/1440-1681.12599   open full text
  • Role of heart rate in the relation between regional body fat and subendocardial viability ratio in women.
    Joaquin U. Gonzales, Omar Hadri.
    Clinical and Experimental Pharmacology and Physiology. July 26, 2016
    Subendocardial viability ratio (SEVR) is a measure of left ventricular function, specifically; it is an index of myocardial perfusion relative to left ventricular workload. Women have lower SEVR than men, partly due to a faster resting heart rate that reduces diastolic time (i.e., time for myocardial perfusion). It is unclear if body fat relates to SEVR, thus the purpose of this study was to examine the relation between body fat and SEVR in women. Twenty‐eight middle‐aged (31–45 years) and 31 older (60–80 years) women were examined. Radial artery applanation tonometry was used to calculate SEVR from a synthesized central aortic pressure wave. Dual‐energy X‐ray absorptiometry was used to assess body composition including fat in the trunk, legs, android and gynoid regions. Body fat was not related (P>.05) with SEVR in older women. In middle‐aged women, all measures of regional fat were correlated with heart rate (range, r=.49–.59, P≤.01) and SEVR (range, r=.43–.53, P≤.01). Android‐to‐gynoid ratio was identified as the strongest predictor (r2=−.26, P<.01) of SEVR among measures of regional fat. Middle‐aged women with lower android‐to‐gynoid fat ratio had higher SEVR (1.96±0.33 vs 1.66±0.20, P=.009) than women with higher fat ratio, even after adjusting for age, height, daily physical activity, and aortic mean pressure (P=.02). Adjusting for heart rate or diastolic time abolished the difference in SEVR between groups (1.80±0.09 vs 1.82±0.09, P=.56). These results suggest that middle‐aged women with a greater distribution of fat in the abdomen have poorer left ventricular function that is dependent on the negative influence of heart rate on diastolic time.
    July 26, 2016   doi: 10.1111/1440-1681.12597   open full text
  • Anorexic response to rapamycin does not appear to involve a central mechanism.
    Hale Z. Toklu, Erin B. Bruce, Yasemin Sakarya, Christy S. Carter, Drake Morgan, Michael K. Matheny, Nataliya Kirichenko, Philip J. Scarpace, Nihal Tümer.
    Clinical and Experimental Pharmacology and Physiology. July 26, 2016
    The authors have previously demonstrated that a low and intermittent peripheral dose of rapamycin (1 mg/kg three times/week) to rats inhibited mTORC1 signalling, but avoided the hyperlipidemia and diabetes‐like syndrome associated with higher doses of rapamycin. The dosing regimen reduced food intake, body weight, adiposity, serum leptin and triglycerides. mTORC1 signalling was inhibited in both liver and hypothalamus, suggesting some of the actions, in particular the decrease in food intake, may be the results of a central mechanism. To test this hypothesis, rapamycin (30 μg/day for 4 weeks) was infused into 23–25‐month‐old F344xBN rats by intracerebroventricular (icv) mini pumps. Our results demonstrated that central infusion did not alter food intake or body weight, although there was a tendency for a decrease in body weight towards the end of the study. mTORC1 signalling, evidenced by decreased phosphorylation of S6 protein at end of 4 weeks, was not activated in liver, hypothalamus or hindbrain. Fat and lean mass, sum of white adipose tissues, brown adipose tissue, serum glucose, insulin and leptin levels remained unchanged. Thus, these data suggest that the anorexic and body weight responses evident with peripheral rapamycin are not the result of direct central action. The tendency for decreased body weight towards the end of study, suggests that there is either a slow transport of centrally administered rapamycin into the periphery, or that there is delayed action of rapamycin at sites in the brain.
    July 26, 2016   doi: 10.1111/1440-1681.12601   open full text
  • CD19+CD20−CD27hi IL‐s10‐producing B cells are overrepresented in R‐CHOP‐treated DLBCL patients in complete remission.
    Huiying Qiu, Junguo Li, Zhenjun Feng, Joanna Yuan, Jie Lu, Xiaoxia Hu, Lei Gao, Shuqing Lv, Jianmin Yang, Lei Chen.
    Clinical and Experimental Pharmacology and Physiology. July 26, 2016
    Treatment of diffuse large B cell lymphoma (DLBCL) with rituximab, an anti‐CD20 monoclonal antibody, has resulted in significantly improved patient responses with longer event‐free intervals and higher overall survival rates. However, since rituximab depletes all CD20‐expressing cells, including noncancerous B cells, the effects of rituximab on the normal immunity of DLBCL patients under remission need to be examined. Here, we observed that DLBCL patients under remission contained significantly lower frequencies of total B cells, with a significantly overrepresented interleukin (IL)‐10‐producing B cell (B10) population in the peripheral blood. Further examination confirmed that a large fraction of B10 cells was CD20−CD27hi plasmablasts, possibly explaining the persistence of B10 cells after R‐CHOP treatment. We also observed that the percentage of B10 cells in DLBCL patients in remission gradually reduced during the first year of achieving complete remission, primarily due to the replenishment of non‐B10 B cells. Despite this, the percentage of B10 cells in DLBCL patients after 1 year of achieving complete remission was still higher than that in controls. CD4+ and CD8+ T cells cocultured with B10‐enriched B cells secreted significantly lower levels of proinflammatory cytokines IFN‐g and TNF‐a, compared to those incubated with B10‐depleted B cells. Together, our data observed a long‐lasting overrepresentation of B10 cells in DLBCL patients under remission. Whether this change could impact on the overall anti‐tumor immunity during remission requires further studies.
    July 26, 2016   doi: 10.1111/1440-1681.12603   open full text
  • Molecular mechanisms for tumour resistance to chemotherapy.
    Shu‐Ting Pan, Zhi‐Ling Li, Zhi‐Xu He, Jia‐Xuan Qiu, Shu‐Feng Zhou.
    Clinical and Experimental Pharmacology and Physiology. July 04, 2016
    Chemotherapy is one of the prevailing methods used to treat malignant tumours, but the outcome and prognosis of tumour patients are not optimistic. Cancer cells gradually generate resistance to almost all chemotherapeutic drugs via a variety of distinct mechanisms and pathways. Chemotherapeutic resistance, either intrinsic or acquired, is caused and sustained by reduced drug accumulation and increased drug export, alterations in drug targets and signalling transduction molecules, increased repair of drug‐induced DNA damage, and evasion of apoptosis. In order to better understand the mechanisms of chemoresistance, this review highlights our current knowledge of the role of altered drug metabolism and transport and deregulation of apoptosis and autophagy in the development of tumour chemoresistance. Reduced intracellular activation of prodrugs (e.g. thiotepa and tegafur) or enhanced drug inactivation by Phase I and II enzymes contributes to the development of chemoresistance. Both primary and acquired resistance can be caused by alterations in the transport of anticancer drugs which is mediated by a variety of drug transporters such as P‐glycoprotein (P‐gp), multidrug resistance associated proteins, and breast cancer resistance protein. Presently there is a line of evidence indicating that deregulation of programmed cell death including apoptosis and autophagy is also an important mechanism for tumour resistance to anticancer drugs. Reversal of chemoresistance is likely via pharmacological and biological approaches. Further studies are warranted to grasp the full picture of how each type of cancer cells develop resistance to anticancer drugs and to identify novel strategies to overcome it.
    July 04, 2016   doi: 10.1111/1440-1681.12581   open full text
  • Comparison of the analgesic effect of patient‐controlled oxycodone and fentanyl for pain management in patients undergoing colorectal surgery.
    Kyeo‐Woon Jung, Hyeon‐Wook Kang, Chan‐Hye Park, Byung‐Hyun Choi, Ji‐Yeon Bang, Soo‐Han Lee, Eun‐Kyung Lee, Byung‐Moon Choi, Gyu‐Jeong Noh.
    Clinical and Experimental Pharmacology and Physiology. July 04, 2016
    Oxycodone is a μ‐opioid receptor agonist and is generally indicated for the relief of moderate to severe pain. The aim of this study was to compare the analgesic efficacy of patient‐controlled oxycodone and fentanyl for postoperative pain in patients undergoing colorectal surgery. Patients scheduled to undergo elective colorectal surgery (n=82) were allocated to receive oxycodone (n=41, concentration of 1 mg/mL) or fentanyl (n=41, concentration of 15 μg/mL) for postoperative pain management. After the operation, pain using a numerical rating scale (NRS), delivery to demand ratio, infused dose of patient‐controlled analgesia (PCA), side effects, and sedation levels were evaluated. Median (25%–75%) cumulative PCA dose of oxycodone group at 48 hours (66.9, 58.4–83.7 mL) was significantly less than that of fentanyl group (80.0, 63.4–103.3 mL, P=.037). Six hours after surgery, the mean (SD) NRS scores of the oxycodone and fentanyl groups were 6.2 (2.4) and 6.8 (1.9), respectively (P=.216). The mean equianalgesic potency ratio of oxycodone to fentanyl was 55:1. The groups did not differ in postoperative nausea, vomiting, and level of sedation. Patient‐controlled oxycodone provides similar effects for pain relief compared to patient‐controlled fentanyl in spite of less cumulative PCA dose. Based on these results, oxycodone can be a useful alternative to fentanyl for PCA in patients after colorectal surgery.
    July 04, 2016   doi: 10.1111/1440-1681.12586   open full text
  • Characterisation of a mouse cerebral microvascular endothelial cell line (bEnd.3) after oxygen glucose deprivation and reoxygenation.
    Jacqueline M Ku, Mohammadali Taher, Kai Yee Chin, Megan Grace, Peter McIntyre, Alyson A Miller.
    Clinical and Experimental Pharmacology and Physiology. July 04, 2016
    Studies have utilised immortalised mouse cerebral endothelial cells (bEnd.3) exposed to oxygen glucose deprivation (OGD) to study blood–brain barrier (BBB) disruption after ischaemia. However, there is a paucity of literature describing the duration of OGD (and reoxygenation [RO]) required to best simulate BBB disruption in vivo. In this study we assessed BBB disruption in bEnd.3 cells after exposure to a range of OGD periods, and also after OGD + RO. Exposure of bEnd.3 monolayers to 4, 6, 16, or 24 hours of OGD resulted in a significant increase in permeability. The hyperpermeability after 16 or 24 hours was associated with decreased expression of tight junction proteins (occludin and claudin‐5). Furthermore, there was a decrease in cell viability and increased expression of the pro‐apoptotic protein, cleaved caspase‐3. Exposure of bEnd.3 monolayers to 1 hour OGD+ 23 hours RO exacerbated hyperpermeability relative to 1 hour OGD, which was associated with decreased expression levels of occludin and ZO‐1, but no change in cell viability or caspase‐3. 4 hours OGD + 23 hours RO exacerbated hyperpermeability, decreased expression levels of tight junction proteins, decreased cell viability, and increased caspase‐3 expression. Thus, bEnd.3 cells exhibit hyperpermeability, a loss of tight junction proteins, and undergo cell death, after exposure to prolonged periods of OGD. Moreover, they exhibit exacerbated hyperpermeability, a loss of tight junction proteins, and increased expression of caspase‐3 after OGD + RO. These findings will facilitate the use of this cell line in studies of BBB disruption and for the testing of therapeutics.
    July 04, 2016   doi: 10.1111/1440-1681.12587   open full text
  • Role of peptide YY in 5‐fluorouracil‐induced reduction of dietary intake.
    Hiroyasu Sakai, Yuki Kai, Kazuhide Takase, Ken Sato, Minami Kimura, Shoko Tabata, Miyabi Yaegashi, Fumiaki Sato, Tetsuro Yomoto, Minoru Narita.
    Clinical and Experimental Pharmacology and Physiology. July 04, 2016
    5‐fluorouracil (5‐FU) is part of the standard care for cancer treatment but is associated with high incidences of appetite loss and reduced food intake, which may contribute to chemotherapy‐induced cachexia (weakness and wasting of tissue). The role of gastrointestinal satiety hormones in chemotherapy‐induced appetite loss has not been intensively investigated. Peptide YY (PYY) and glucagon‐like peptide (GLP)‐1 are important signals of gastrointestinal satiety, so this study examined the roles of these gut hormones in 5‐FU‐induced reduction of dietary intake. Mice were given 5‐FU (50 mg/kg, intraperitoneal [i.p.]) every day for 4 consecutive days. Gene expression levels of proglucagon (Pro‐Gcg), a precursor of GLP‐1, and PYY in the colon were examined by real‐time RT‐PCR. Serum levels of GLP‐1 and PYY were measured by enzyme‐linked immunosorbent assay. Some mice were pretreated with the GLP‐1 receptor antagonist exendin9–39 (1 mg/kg) or the neuropeptide Y type 2 (NPY2) receptor antagonist BIIE0246 (2 mg/kg) via the i.p. route 30 minutes before 5‐FU administration. Mice receiving 5‐FU exhibited a significant reduction in food intake that was correlated with body weight loss. These mice also showed significantly enhanced expression levels of mRNAs encoding pro‐GLP‐1 and PYY in the transverse and distal colon as well as elevated serum concentrations of GLP‐1 and PYY compared to vehicle‐treated controls. The 5‐FU‐induced reduction in food intake was attenuated by BIIE0246 but not by exendin9–39. These data suggest that administration of a NPY2 receptor antagonist may be effective for attenuating the anorexia caused by 5‐FU chemotherapy.
    July 04, 2016   doi: 10.1111/1440-1681.12588   open full text
  • Cytoprotective effects of endothelin‐1 on mesenchymal stem cells: an in vitro study.
    Mona Pourjafar, Massoud Saidijam, Kamran Mansouri, Sara Malih, Tayebeh Ranjbar Nejad, Nooshin Shabab, Rezvan Najafi.
    Clinical and Experimental Pharmacology and Physiology. July 04, 2016
    Stem cell‐based therapies is a promising approach for regenerative therapy in various diseases. Some obstacles remain to be solved before clinical application of the cell therapy is realized, including increasing the survival of transplanted stem cells, reducing loss of transplanted cells, and maintaining adequate vascular supply. Recently, stem cell preconditioning with chemical and pharmacological agents has been shown to increase therapeutic efficacy. The present study investigated the effect of endothelin‐1 (ET‐1) on survival, angiogenesis, and migration of mesenchymal stem cells (MSCs), in vitro. MSCs were treated with various concentrations of ET‐1 and the expression of cyclooxygenase‐2 (COX‐2), hypoxia‐inducible factor‐1 (HIF‐1), C‐X‐C chemokine receptor type 4 (CXCR4), C‐C chemokine receptor type 2 (CCR2), vascular endothelial growth factor (VEGF), angiopoietin‐2 (Ang‐2), angiopoietin‐4 (Ang‐4) and matrix metalloproteinase‐2 (MMP‐2) were examined. Caspase 3 activity and prostaglandin E2 (PGE2) were determined by ELISA assay. MSCs migration and tube formation potential were assessed using scratch test and three dimensional vessel formation assay. ET‐1 enhanced the MSCs viability. In ET‐1‐ treated MSCs, expression of COX‐2, HIF‐1, CXCR4, CCR2, VEGF, Ang‐2, Ang‐4 and MMP‐2 were increased compared to control groups. Elevation of all these genes were reversed by celecoxib (50 μmol/L), a selective COX‐2 inhibitor. PGE2 generation, MSCs migration and tube formation were enhanced by ET‐1 conditioning, whereas caspase‐3 activity was reduced in these cells, compared to the control group. The results presented here reveal that preconditioning of MSCs with ET‐1 has strong cytoprotective effects through activation of survival signalling molecules and trophic factors.
    July 04, 2016   doi: 10.1111/1440-1681.12590   open full text
  • Deferoxamine alleviates liver fibrosis induced by CCl4 in rats.
    Aya Mohammed, Ekram N Abd Al Haleem, Wesam M El‐Bakly, Ebtehal El‐Demerdash.
    Clinical and Experimental Pharmacology and Physiology. July 04, 2016
    Several chronic liver diseases can lead to the occurrence of hepatic fibrosis through the accumulation of iron, which causes induction of oxidative stress and consequently activation of fibrogenesis. The present study was designed to investigate the potential antifibrotic and anti‐oxidant effects of deferoxamine (DFO), a well‐known iron chelator in an experimental rat model of liver injury using carbon tetrachloride (CCl4). First, the potential effective dose of DFO was screened against CCl4‐induced acute hepatotoxicity. Then, rats were co‐treated with DFO (300 mg/kg, i.p.) for 6 weeks starting from the third week of CCl4 induction of chronic hepatotoxicity. Liver function was assessed in addition to histopathological examination. Furthermore, oxidative stress and fibrosis markers were assessed. It was found that treatment of animals with DFO significantly counteracted the changes in liver function; histopathological lesions and hepatic iron deposition that were induced by CCl4. DFO also significantly counteracted the CCl4‐induced lipid peroxidation increase and reduction in antioxidant activities of superoxide dismutase and glutathione peroxidase enzymes. In addition, DFO ameliorated significantly liver fibrosis markers including hydroxyproline, collagen accumulation, and the expression of the hepatic stellate cells (HSCs) activation marker; alpha smooth muscle actin and transforming growth factor‐beta (TGF‐β). Together, these findings indicate that DFO possesses a potent antifibrotic effect due to its antioxidant properties that counteracted oxidative stress and lipid peroxidation and restored antioxidant enzymes activities as well as reducing HSCs activation and fibrogenesis.
    July 04, 2016   doi: 10.1111/1440-1681.12591   open full text
  • Retracted: ‘Ergothioneine represses inflammation and dysfunction in human endothelial cells exposed to oxidized low‐density lipoprotein’ by Lunna Saing, Yu‐Chi Wei and Chih‐Jen Tseng.

    Clinical and Experimental Pharmacology and Physiology. June 22, 2016
    The above article from Clinical and Experimental Pharmacology and Physiology, published as an Accepted Article online on 10 February 2015 in Wiley Online Library (, has been retracted by agreement between the authors, the Journal Editor‐in‐Chief, Jun‐Ping Liu, and John Wiley & Sons Australia, Ltd. The retraction has been agreed as the article was submitted without agreement of all co‐authors. Reference Saing L, Wei Y‐C, Tseng C‐J. Ergothioneine represses inflammation and dysfunction in human endothelial cells exposed to oxidized low‐density lipoprotein. Clin Exp Pharmacol Physiol 2015; DOI: 10.1111/1440‐1681.12374
    June 22, 2016   doi: 10.1111/1440-1681.12374   open full text
  • Chronic mercury exposure at different concentrations produces opposed vascular responses in rat aorta.
    BF Azevedo, MR Simões, J Fiorim, T Botelho, JK Angeli, JVA Vieira, MJ Alonso, M Salaices, L Santos, DV Vassallo.
    Clinical and Experimental Pharmacology and Physiology. June 22, 2016
    Mercury chloride exposure for 30 days decreases NO bioavailability and increases oxidative stress. However, the mechanisms underlying the effects of mercury on the cardiovascular system are not completely understood, and it is not known if they are dose‐dependent or if some concentrations have no harmful effects. Thus, we investigated the effects of chronic exposure to doses low (half) and high (2.5‐fold higher) than that needed to obtain 29 nmol/L of HgCl2 on the vascular function. Three‐month‐old male Wistar rats received intramuscular (i.m.) HgCl2 for 30 days and were divided in three groups: lower (Low Hg); higher (High Hg); and saline was used as the control. High Hg exposure increased the contractile response to phenylephrine (PHE) in aortic rings, but Low Hg reduced it. The hyporesponsiveness in the Low Hg rats was blunted by endothelial denudation and NOS inhibition with l‐NAME (100 μmol/L). The phosphorylated‐eNOS/eNOS protein ratio increased in the aortas of Low Hg rats. In the High Hg group, endothelial denudation increased the PHE‐induced contractions, while l‐NAME had no effects and indomethacin (10 μmol/L), losartan (10 μmol/L) and apocynin (30 μmol/L) reduced this response. In the High Hg group, protein levels of the NADPH oxidase subunit gp91phox and cyclooxygenase‐2 increased. Our results support previous suggestions that High Hg increases oxidative stress that might activate an inflammatory cascade and the renin‐angiotensin system. However, very low Hg concentrations below the level considered safe still reduced vascular reactivity, suggesting the need for special attention to continuous exposure as a putative cause of increased cardiovascular risk.
    June 22, 2016   doi: 10.1111/1440-1681.12578   open full text
  • The effect of enoxaparin on seroma and mesh‐tissue adhesion in a hernia model.
    Ziya T Ozkececi, Yucel Gonul, Afra Karavelioglu, Mehmet F Bozkurt, Emre Kacar, Ahmet Bal, Mustafa Ozsoy, Ozan Turamanlar, Bahadir Celep.
    Clinical and Experimental Pharmacology and Physiology. June 22, 2016
    The aim of this study was to investigate whether enoxaparin (ENX) administration would increase seroma risk and worsen mesh tissue recovery in an experimental rat hernia repair model. Fifty‐six adult male Wistar–Albino rats were included in the study. Rats were equally and randomly separated into seven groups: Group 1, Control, only subcutaneous dissection was performed; group 2, Sham, Hernia defect was primary sutured; Group 3, Prolene mesh; Group 4, Dual mesh; Group 5, ENX + Sham; Group 6, ENX + Prolene mesh; Group 7, ENX + Dual mesh. ENX was subcutaneously injected at a dose of 180 U/kg per day for 7 days. Rats were killed after the amount of subcutaneous seroma was determined by ultrasound on day 7 following the surgical procedure. Mesh‐tissue healing was evaluated using histopathological and immunohistochemical (CD31) staining methods. The mean seroma amount significantly increased in Groups 5–7 compared to Groups 2–4. CD31 immunostaining showed a reduction in neovascularization in Groups 6 and 7, compared to Groups 3 and 4. Neovascularization decreased and hemorrhage, necrosis and oedema findings remarkably increased in Groups 6 and 7, when compared to Groups 3 and 4. Fibroblastic activity and inflammation were more prominent in Groups 3 and 4. It should be kept in mind that ENX interferes with inflammation, which is desired in the early period of healing and leads to an increase in overall seroma amount with anti‐coagulant effects, which in turn may disrupt wound healing and mesh‐tissue adhesions, as was indicated in our study.
    June 22, 2016   doi: 10.1111/1440-1681.12582   open full text
  • Downregulation of toll‐like receptor 4 and IL‐6 following irradiation of the rat urinary bladder.
    D Giglio, C Wasén, J Mölne, D Suchy, J Swanpalmer, J Jabonero Valbuena, G Tobin, L Ny.
    Clinical and Experimental Pharmacology and Physiology. June 22, 2016
    The pathophysiology behind radiation cystitis is poorly understood. Here we investigated whether bladder irradiation affects the immune system of the rat urinary bladder. Female rats were sedated and exposed to one single radiation dose of 20 Gy or only sedated (controls) and killed 16 h to 14 days later. Rats were placed in a metabolic cage at 16 h, 3 days, 7 days and 14 days following bladder irradiation. The urinary bladders were harvested and analysed with qPCR, immunohistochemistry and/or Western blot for the expression of interferon (IFN)‐γ, interleukin (IL)‐1β, IL‐2, IL‐4, IL‐5, IL‐6, IL‐10, IL‐13, nitric oxide synthases (eNOS, iNOS and nNOS), tumour necrosis factor (TNF)‐α and toll‐like receptor 4 (TLR4). Urine was collected and analysed for IL‐6 and nitrite (reflecting nitric oxide activity) with ELISA and the Griess reaction, respectively. Irradiation increased bladder frequency and decreased voiding volumes 14 days following bladder irradiation. Bladder irradiation increased the expression of IL‐10 and collagen in the bladder, while TLR4 and IL‐6 expressions were decreased in the urothelium concomitantly with a decrease in mast cells in the submucosa and urine levels of IL‐6 and nitrite. The present findings show that bladder irradiation leads to urodynamic changes in the bladder and may suppress important immunoregulatory pathways in the urinary bladder.
    June 22, 2016   doi: 10.1111/1440-1681.12583   open full text
  • An investigation of the influence of patient‐related factors and comedications on lamotrigine clearance in patients with epilepsy.
    André Oliveira Baldoni, Priscila Freitas‐Lima, Flávia Isaura Santi Ferreira, Edson Zangiacomi Martinez, Regina Helena Costa Queiroz, Americo Ceiki Sakamoto, Veriano Alexandre, Emilio Perucca, Leonardo Regis Leira Pereira.
    Clinical and Experimental Pharmacology and Physiology. June 22, 2016
    Lamotrigine (LTG) is one of the most widely used antiepileptic drugs. Confusion still exists in the literature as to the relative influence of age, body weight, and concomitant drug therapy on LTG pharmacokinetics. So, the objective of this study is to evaluate the influence of patient‐related factors and comedication on LTG apparent oral clearance (CL/F). A therapeutic drug‐monitoring database was used to identify steady‐state plasma LTG concentrations in 210 patients. LTG CL/F values were calculated for each patient according to the equation CL/F (L/h per kg) = LTG daily dose (mg/kg)/Css (steady state concentration) (mg/L) × 24 h. A linear‐regression model was used to assess the influence of gender, dose, age, and body weight in LTG CL/F. The influence of comedication on LTG CL/F was investigated by applying the Bonferroni post‐test. The lowest LTG CL/F was found in patients comedicated with valproate (VPA) (mean, 0.0183 L/h per kg), followed by patients receiving VPA + enzyme inducers (0.0271 L/h per kg), patients on LTG monotherapy (0.0298 L/h per kg) and patients comedicated with enzyme inducers (0.056 L/h per kg) LTG CL/F correlated significantly with LTG dose (P < 0.01), but showed no significant relationship with gender, weight, and age. LTG CL/F is influenced by the type of antiepileptic comedication. The correlation with dose may be a spurious finding related to the fact that physicians, in adjusting dosage according to clinical response, are more likely to use larger doses in patients with high clearance values.
    June 22, 2016   doi: 10.1111/1440-1681.12584   open full text
  • Role of tumour necrosis factor‐a in the regulation of T‐type calcium channel current in HL‐1 cells.
    Fang Rao, Yu‐mei Xue, Wei Wei, Hui Yang, Fang‐zhou Liu, Shao‐xian Chen, Su‐juan Kuang, Jie‐ning Zhu, Shu‐lin Wu, Chun‐yu Deng.
    Clinical and Experimental Pharmacology and Physiology. June 22, 2016
    Increasing evidence indicates that inflammation contributes to the initiation and perpetuation of atrial fibrillation (AF). Although tumour necrosis factor (TNF)‐α levels are increased in patients with AF, the role of TNF‐α in the pathogenesis of AF remains unclear. Besides L‐type Ca2+ currents (ICa,L), T‐type Ca2+ currents (ICa,T) also plays an important role in the pathogenesis of AF. This study was designed to use the whole‐cell voltage‐clamp technique and biochemical assays to explore if TNF‐α is involved in the pathogenesis of AF through regulating ICa,T in atrial myocytes. It was found that compared with sinus rhythm (SR) controls, T‐type calcium channel (TCC) subunit mRNA levels were decreased, while TNF‐α expression levels were increased, in human atrial tissue from patients with AF. In murine atrial myocyte HL‐1 cells, after culturing for 24 h, 12.5, 25 and 50 ng/mL TNF‐α significantly reduced the protein expression levels of the TCC α1G subunit in a concentration‐dependent manner. The peak current was reduced by the application of 12.5 or 25 ng/mL TNF‐α in a concentration‐dependent manner (from −15.08 ± 1.11 pA/pF in controls to −11.89 ± 0.83 pA/pF and −8.54 ± 1.55 pA/pF in 12.5 or 25 ng/mL TNF‐α group respectively). TNF‐α application also inhibited voltage‐dependent inactivation of ICa,T, shifted the inactivation curve to the left. These results suggest that TNF‐α is involved in the pathogenesis of AF, probably via decreasing ICa,T current density in atrium‐derived myocytes through impaired channel function and down‐regulation of channel protein expression. This pathway thus represents a potential pathogenic mechanism in AF.
    June 22, 2016   doi: 10.1111/1440-1681.12585   open full text
  • The role of Toll‐like receptor 4 (TLR4) in cardiac ischaemic‐reperfusion injury, cardioprotection and preconditioning.
    S M Lee, M Hutchinson, D A Saint.
    Clinical and Experimental Pharmacology and Physiology. June 01, 2016
    Cardiac ischaemic reperfusion injury (IRI) remains the primary cause of mortality throughout the developed world. Molecular mechanisms underlying IRI are complex and are often interlinked with each other driving a synergistic response. Toll‐like receptor 4 (TLR4), an immunosurveillance receptor, is known to enhance tissue injury during IRI by enhancing the inflammatory response. The release of endogenous components during IRI bind onto TLR4 leading to the activation of multiple signalling kinases. Once this event occurs these proteins are defined as DAMPs (danger associated molecular patterns molecules) or alarmins. Examples include heat shock proteins, high mobility group box one (HMGB1) and extracellular matrix proteins, all of which are involved in IRI. However, literature in the last two decades suggests that transient stimulation of TLR4 may suppress IRI and thus improve cardiac recovery. Furthermore, it remains to be seen what role TLR4 plays during ischaemic‐preconditioning where acute bouts of ischaemia, preceding a harmful bout of ischaemic‐reperfusion, is cardioprotective. The other question which also needs to be considered is that if transient TLR4 signalling drives a preconditioning response then what are the ligands which drive this? Hence the second part of this review explores the possible TLR4 ligands which may promote cardioprotection against IRI. This article is protected by copyright. All rights reserved.
    June 01, 2016   doi: 10.1111/1440-1681.12602   open full text
  • β‐asarone and levodopa co‐administration increase striatal dopamine level in 6‐hydroxydopamine induced rats by modulating P‐glycoprotein and tight junction proteins at the blood‐brain barrier and promoting levodopa into the brain.
    Liping Huang, Minzhen Deng, Yuping He, Shiyao Lu, Ruanxin Ma, Yongqi Fang.
    Clinical and Experimental Pharmacology and Physiology. May 04, 2016
    Levodopa (L‐dopa) is widely considered as one of the most effective drug constituents in the treatment of Parkinson's disease (PD), but the blood‐brain barrier (BBB) permeability of L‐dopa is <5%, which causes low efficacy. Neuroprotective effects of β‐asarone on 6‐hydroxydopamine (6‐OHDA)‐induced PD rats were demonstrated by our previous studies. Co‐administration of β‐asarone and L‐dopa has not been explored until being investigated on PD rats in this study. PD rats were divided into four groups: untreated, L‐dopa‐treated, β‐asarone‐treated and co‐administered‐treated groups. All of the treatments were administered to the rats twice per day for 30 days. The L‐dopa, dopamine (DA), 3,4‐dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA), S100β and neuron‐specific enolase (NSE) levels were subsequently determined. The P‐glycoprotein (P‐gp), zonula occludens‐1 (ZO‐1), claudin‐5, occludin and actin expression was also assessed in cortex. Changes in BBB ultrastructure were observed using transmission electron microscopy. Our results showed that the co‐administered treatment increased levels of L‐dopa, DA, DOPAC and HVA in striatum, and S100β in plasma, but down‐regulated NSE, P‐gp, ZO‐1, occludin, actin and claudin‐5 in cortex. Crevices were observed between capillary endothelial cells at intercellular tight junction of the striatum in co‐administered‐treated group, while the endothelial cells in untreated group were tightly jointing each other. In addition, the correlations of L‐dopa or DA and P‐gp or tight junction proteins respectively were significantly negative in co‐administered‐ and β‐asarone‐treated groups. These findings suggest that co‐administered treatment may enhance the L‐dopa BBB permeability and attenuate brain injury, which may be beneficial to PD treatment.
    May 04, 2016   doi: 10.1111/1440-1681.12570   open full text
  • Silencing of hypoxia inducible factor‐1α by RNA interference inhibits growth of SK‐NEP‐1 Wilms tumour cells in vitro, and suppresses tumourigenesis and angiogenesis in vivo.
    Bo Shi, Ying Li, Xiuli Wang, Yi Yang, Dan Li, Xin Liu, Xianghong Yang.
    Clinical and Experimental Pharmacology and Physiology. May 04, 2016
    Wilms tumour is the most common tumour of the pediatric kidney. Elevation of hypoxia‐inducible factor 1α (HIF‐1α) has been detected in 93% to 100% of human Wilms tumour specimens, suggesting a potential value of HIF‐1α as a therapeutic target for Wilms tumour. In the present study, a stable HIF‐1α‐silenced Wilms tumour cell strain was established by introducing HIF‐1α short‐hairpin RNA (shRNA) into SK‐NEP‐1 cells. Silencing of HIF‐1α significantly reduced single‐cell growth capacity, suppressed proliferation and arrested cell cycle of SK‐NEP‐1 cells. In addition, reduction of HIF‐1α expression induced apoptosis in SK‐NEP‐1 cells, which was accompanied by increased levels of cleaved caspase‐3, cleaved poly (ADP‐ribose) polymerase (PARP) and Bax as well as downregulation of Bcl‐2 in the cells. Furthermore, when inoculated subcutaneously in nude mice, HIF‐1α‐silenced SK‐NEP‐1 cells displayed retarded tumour growth and impaired tumour angiogenesis. In summary, the findings of this study suggest that HIF‐1α plays a critical role in the development of Wilms tumour, and it may serve as a candidate target of gene therapy for Wilms tumour.
    May 04, 2016   doi: 10.1111/1440-1681.12575   open full text
  • miR‐410 suppresses the expression of interleukin‐6 as well as renal fibrosis in the pathogenesis of lupus nephritis.
    Dongmei Liu, Na Zhang, Jing Zhang, Haiyan Zhao, Xiaofei Wang.
    Clinical and Experimental Pharmacology and Physiology. May 04, 2016
    Lupus nephritis (LN) is a highly complex autoimmune disease caused by systemic lupus erythematosus (SLE). MicroRNAs (miRNAs) play a vital role in the pathogenesis of SLE. Previously, a total of 29 miRNAs were identified to be down‐regulated in SLE patients, in which miR‐410 was likely to be involved in the signalling transduction pathways in regulating the pathogenesis of SLE. The purpose of this study was to investigate the role of miR‐410 in LN and to find out whether miR‐410 regulates the expression of interleukin (IL)‐6 and fibrosis in LN. It was found that the expression level of miR‐410 in kidney tissue of MRL/lpr mice was decreased compared to that in BALB/C mice, whereas the level of IL‐6 was overexpressed in MRL/lpr mice. Luciferase assay showed that miR‐410 binds directly to the 3′ untranslated region (UTR) of IL‐6, with the results showing that overexpression of miR‐410 significantly decreased the expression level of IL‐6 in SV40MES13 cells. Moreover, overexpression of miR‐410 significantly reduced the expression levels of fibrosis factors such as transforming growth factor‐β1 (TGF‐β1) and collagen I/III in SV40MES13 cells; Inhibition of the expression of miR‐410 with miR‐410 inhibitor resulted in increased levels of IL‐6 as well as fibrosis factors. The results identify that miR‐410, as a novel and critical factor in the pathogenesis of LN, decreases IL‐6 expression by binding directly to the 3′UTR and suppresses fibrosis through down‐regulation of TGF‐β1 in SV40MES13 cells. Our study brings new insight into understanding the complex mechanisms involved in the pathogenesis of lupus disease.
    May 04, 2016   doi: 10.1111/1440-1681.12576   open full text
  • The initial fall in arterial pressure evoked by endotoxin is mediated by the ventrolateral periaqueductal gray.
    William R Millington, M Sertac Yilmaz, Carlos Feleder.
    Clinical and Experimental Pharmacology and Physiology. May 04, 2016
    This study tested the hypothesis that the initial fall in arterial pressure evoked by lipopolysaccharide (LPS) is mediated by the ventrolateral column of the midbrain periaqueductal gray region (vlPAG). To test this hypothesis, the local anaesthetic lidocaine (2%; 0.1 μL, 0.2 μL or 1.0 μL), the delta opioid receptor antagonist naltrindole (2 nmol) or saline was microinjected into the vlPAG of isoflurane‐anaesthetized rats bilaterally and LPS (1 mg/kg) or saline was administered intravenously 2 min later. Both lidocaine and naltrindole inhibited LPS‐evoked hypotension significantly but did not affect arterial pressure in saline‐treated control animals. Neither lidocaine nor naltrindole altered heart rate significantly in either LPS‐treated or control animals. Microinjection of lidocaine or naltrindole into the dorsolateral PAG was ineffective. These data indicate that the vlPAG plays an important role in the initiation of endotoxic hypotension and further show that delta opioid receptors in the vlPAG participate in the response.
    May 04, 2016   doi: 10.1111/1440-1681.12573   open full text
  • Inhibition of platelet aggregation by vanilloid‐like agents is not mediated by transient receptor potential vanilloid‐1 channels or cannabinoid receptors.
    Safa Almaghrabi, Dominic Geraghty, Kiran Ahuja, Murray Adams.
    Clinical and Experimental Pharmacology and Physiology. May 04, 2016
    Vanilloid‐like agents, including capsaicin, N‐arachidonoyl‐dopamine and N‐oleoyldopamine inhibit platelet aggregation, however little is known about the precise mechanism(s) of action. The authors have previously shown that blocking of the capsaicin receptor, transient receptor potential vanilloid‐1 (TRPV1), does not interfere with capsaicin action during adenosine diphosphate (ADP)‐induced aggregation. This research is extended to investigate the effect of these vanilloid‐like‐agents on platelet count, and to test whether the effect of these agents is mediated through TRPV1 and/or cannabinoid (CB1 and CB2) receptors in the presence of other agonists, including collagen and arachidonic acid. Incubation of platelets with each of the individual vanilloids, or with receptor antagonists of TRPV1 (SB452533), CB1 (AM251) and CB2 (AM630), for up to 2 h did not significantly affect the platelet count. Similarly, the effect of individual vanilloids on the inhibition of platelet aggregation was not significantly different in the presence of receptor agonists compared to control, irrespective of the agonist used, suggesting that the inhibitory effect of vanilloids on platelet aggregation is independent of TRPV1, CB1 and CB2 receptors. Further research on the antiplatelet activity of vanilloids should focus on mechanisms other than those associated with vanilloid receptors.
    May 04, 2016   doi: 10.1111/1440-1681.12569   open full text
  • An update on the pharmacokinetics and pharmacodynamics of alisertib, a selective Aurora kinase A inhibitor.
    Cameron T Durlacher, Zhi‐Ling Li, Xiao‐Wu Chen, Zhi‐Xu He, Shu‐Feng Zhou.
    Clinical and Experimental Pharmacology and Physiology. May 04, 2016
    Human Aurora kinases, including Aurora kinase A (AURKA), B (AURKB), and C (AURKC), play an essential role in mitotic events such as monitoring of the mitotic checkpoint, creation of bipolar mitotic spindle and alignment of centrosomes on it, also regulating centrosome separation, bio‐orientation of chromosomes and cytokinesis. AURKA and AURKB are key regulators of mitosis and centrosome via polymerizing microfilaments and controlling chromatid segregation. In particular, AURKA plays critical roles in the regulation of mitotic entry, centrosome function, bipolar spindle assembly, and chromosome segregation. AURKA has been found to be overexpressed in various solid and haematological cancers and has been linked with poor prognosis. Its important role in cancer initiation, growth, and metastasis has brought the focus to search for potent and selective AURKA inhibitors for cancer treatment. MLN8237, also known as alisertib, is one selective AURKA inhibitor that has shown remarkable anticancer effects in preclinical studies. Alisertib exhibits favourable pharmacokinetic properties. Alisertib has generally showed good partial response rates of 4–52% and good safety profiles in Phase I and II trials when it is solely administered as well as combined with cytotoxic chemotherapeutic drugs. Recently, the multicentre, randomized Phase III study of alisertib in patients with relapsed or refractory peripheral T‐cell lymphoma has been discontinued due to unsatisfactory efficacy. The low risk of side effects, accessibility, and effectiveness of alisertib makes it a new promising anticancer therapy and further mechanistic and clinical studies are warranted.
    May 04, 2016   doi: 10.1111/1440-1681.12571   open full text
  • Low‐dose metronomic chemotherapy with cisplatin enhanced immunity in a murine model of ectopic cervical cancer.
    Ching‐Chou Tsai, Jian‐Tai Qiu, Chih‐Wen Tseng, Yi‐Chiang Hsu.
    Clinical and Experimental Pharmacology and Physiology. November 06, 2015
    Previous researchers have claimed that metronomic low‐dose/dense chemotherapy can enhance the therapeutic effectiveness of cisplatin treatment in the control of cancer. Therefore, the aim of this study was to explore the effectiveness of metronomic drug delivery with regards to its effects on adaptive immunity in a murine model of ectopic cervical cancer. The effectiveness of long‐term low‐dose/dense cisplatin treatment in HPV E7‐expressing TC‐1 cells was evaluated via morphological observations. Tumour mass and survival curves were used to determine the antitumour effect against E7‐expressing tumours. After experimental mice had been treated with low‐dose/dense cisplatin therapy, flow cytometry was used to measure the expression of MHC class I surface antigens on cultured TC‐1 cells. Splenocytes expressing both IFN‐γ and CD8 responsible for E7 antigens and the Treg population were also quantified using flow cytometry. The results indicated that in vivo treatment with metronomic cisplatin suppressed the growth of cultured TC‐1 cells. We also observed an increase in the number of splenocytes expressing both IFN‐γ and CD8 responsible for E7 antigens and the Treg population. These results support previous reports that metronomic low‐dose/dense cisplatin chemotherapy is an effective treatment against ectopic cervical cancer with E7‐expression. This article is protected by copyright. All rights reserved.
    November 06, 2015   doi: 10.1111/1440-1681.12515   open full text
  • Ferulic acid improves lipid and glucose homeostasis in high‐fat diet‐induced obese mice.
    Jarinyaporn Naowaboot, Pritsana Piyabhan, Narongsuk Munkong, Wason Parklak, Patchareewan Pannangpetch.
    Clinical and Experimental Pharmacology and Physiology. November 06, 2015
    Ferulic acid (FA) is a plant phenolic acid having several pharmacological effects including antihyperglycemic activity. Thus, the objective of this study was to investigate the effect of FA on glucose and lipid metabolism in the high‐fat diet (HFD)‐induced obese mice. The ICR mice were fed an HFD (45 kcal% fat) for 16 weeks. At the 9th week of induction, the obese mice were orally administered with daily FA doses of 25 and 50 mg/kg for the next eight weeks. The results showed that FA significantly reduced the elevated blood glucose and serum leptin levels, lowered the insulin resistance, and increased the serum adiponectin level. Moreover, serum lipid level, and liver cholesterol and triglyceride accumulations were also reduced. The histological examination showed a clear evidence of a decrease in the lipid droplets in liver tissues and smaller size of fat cells in the adipose tissue in the obese mice treated with FA. Interestingly, FA reduced the expression of hepatic lipogenic genes such as sterol regulatory element‐binding protein 1c (SREBP1c), fatty acid synthase (FAS), and acetyl‐CoA carboxylase (ACC). It could also up‐regulate hepatic carnitine palmitoyltransferase 1a (CPT1a) gene and peroxisome proliferator‐activated receptor alpha (PPARα) proteins. The FA treatment was also found to suppress the protein expressions of hepatic gluconeogenic enzymes, phosphoenolpyruvate carboxylase (PEPCK) and glucose‐6‐phosphatase (G6Pase). In conclusion, our findings demonstrated that FA improved the glucose and lipid homeostasis in HFD‐induced obese mice probably via modulating the expression of lipogenic and gluconeogenic genes in liver tissues. This article is protected by copyright. All rights reserved.
    November 06, 2015   doi: 10.1111/1440-1681.12514   open full text
  • Effects of captopril, telmisartan, and bardoxolone methyl (CDDO‐Me) in ischemia reperfusion‐induced acute kidney injury in rats: an experimental comparative study.
    Cengiz Kocak, Emel Fatma Kocak, Raziye Akcilar, Zeynep Bayat, Bekir Aras, Mehmet Huseyin Metineren, Mehmet Yucel, Hasan Simsek.
    Clinical and Experimental Pharmacology and Physiology. October 30, 2015
    Renal ischemia‐reperfusion (IR) injury is one of the most common causes of acute kidney injury. We investigated the effects of captopril (CAP), telmisartan (TEL), and bardoxolone methyl (BM) in animals with renal IR injury. Adult male Wistar‐Albino rats were divided into six groups: control, vehicle, IR, IR with CAP, IR with TEL, and IR with BM. Before IR was induced, drugs were administered by oral gavage. After a 60‐min ischemia and a 120‐min reperfusion period, bilateral nephrectomies were performed. Serum urea, creatinine, neutrophil gelatinase‐associated lipocalin (NGAL) levels, tissue total oxidant status (TOS), total antioxidant status (TAS), total thiol (TT), asymmetric dimethylarginine (ADMA) levels, superoxide dismutase (SOD) activity, and glutathione peroxidase (GSH‐Px) activity were measured. Tissue mRNA expression levels of peroxisome proliferator‐activated receptor‐ɣ (PPAR‐ɣ), nuclear factor erythroid 2‐related factor 2 (Nrf2), and nuclear factor kappa‐light‐chain‐enhancer of activated B cells (NF‐κB) were analyzed. In addition, renal tissues were evaluated histopathologically and immunohistochemically. All tested drugs reduced renal damage, apoptosis, urea, creatinine, NGAL, TOS, nitric oxide (NO), and ADMA levels, NF‐κB, inducible nitric oxide synthase (iNOS), and endothelin‐1 (ET‐1) expressions (P < 0.001). All tested drugs increased SOD activity, GSH‐Px activity, TAS levels, TT levels, endothelial nitric oxide synthase (eNOS) expression, dimethylarginine dimethylaminohydrolases (DDAHs) expression, Nrf2 expression, and PPAR‐ɣ expression (P < 0.001, P < 0.003). Our results suggest that CAP, TEL, and BM pretreatment could reduce renal IR injury via anti‐inflammatory, antioxidant, and anti‐apoptotic effects. This article is protected by copyright. All rights reserved.
    October 30, 2015   doi: 10.1111/1440-1681.12511   open full text
  • Curcumin alleviates glucocorticoid‐induced osteoporosis by protecting osteoblasts from apoptosis in vivo and in vitro.
    Zhiguang Chen, Jinqi Xue, Tao Shen, Gen Ba, Dongdong Yu, Qin Fu.
    Clinical and Experimental Pharmacology and Physiology. October 30, 2015
    Curcumin, an active component of the rhizomes of Curcumin longa L., possess broad anti‐inflammation and anti‐cancer activities. Curcumin was previously reported to be capable of protecting ovariectomized rats against osteoporosis. However, the effect of curcumin on glucocorticoid‐induced osteoporosis (GIO) is not clear yet. The present study investigated the effects of curcumin on dexamethasone (Dex)‐induced osteoporosis in vivo and Dex‐induced osteoblast apoptosis in vivo and in vitro. The GIO rat model was induced by subcutaneous injection of Dex for 60 days and verified to be successful as evidence by the significantly decreased bone mineral density (BMD) using dual X‐ray absorptiometry. Subsequently, curcumin administration (100 mg/kg) for 60 days obviously increased BMD and bone‐alkaline phosphatase, decreased carboxy‐terminal collagen cross links, enhanced bone mechanical strength, and improved trabecular microstructure, thereby alleviating Dex‐induced osteoporosis. Mechanically, curcumin remarkably reserved Dex‐induced femoral osteoblast apoptosis in vivo. In cultured primary osteoblasts, pretreatment with curcumin concentration‐dependently decreased the number of Dex‐induced apoptotic osteoblasts by down‐regulating the ratio of Bax/Bcl‐2 as well as the levels of cleaved caspase‐3 and cleaved PARP. Moreover, curcumin pretreatment activated extracellular signal regulated kinase (ERK) signaling in Dex‐induced osteoblasts by up‐regulating the expression level of p‐ERK1/2. Taken together, our study demonstrated that curcumin could ameliorate GIO by protecting osteoblasts from apoptosis, which was possibly related to the activation of ERK pathway. The results suggest that curcumin may be a promising drug for prevention and treatment of GIO. This article is protected by copyright. All rights reserved.
    October 30, 2015   doi: 10.1111/1440-1681.12513   open full text
  • Sustained increase in platelet aggregation after the cessation of clopidogrel.
    Nina Djukanovic, Zoran Todorovic, Danijela Zamaklar‐Trifunovic, Dragana Protic, Boris Dzudovic, Miodrag Ostojic, Slobodan Obradovic.
    Clinical and Experimental Pharmacology and Physiology. October 30, 2015
    We have shown that abrupt cessation of one‐year clopidogrel treatment was not associated with thrombotic events in prospective, multicenter study that enrolled 200 patients subjected to coronary stent implantation and treated with aspirin + clopidogrel one year after the stent placement. The aim of this study was to investigate the causes of sustaining increase of platelet aggregability, considering that the values of platelet aggregation stimulated with ADP+PGE1 (ADPHS values) significantly increased 10‐90 days after the cessation of clopidogrel. Values of platelet aggregation induced by thrombin receptor activating peptide (TRAP values) and arachidonic acid (ASPI values) were divided into quartiles on the basis of ADPHS values 10 days after stopping clopidogrel (ADPHS10). There was a significant difference between TRAP values divided into quartiles according to ADPHS10, 10, 45 and 90 days after stopping clopidogrel (p < 0.001, all), and ASPI values across the same quartiles 10 and 45 days after the cessation of clopidogrel (p = 0.028 and 0.003). Results of our study indicate that patients with early pronounced rebound phenomenon to clopidogrel termination have a long‐term (at least 90 days) increased platelet aggregation to other agonists such as thrombin related activated protein and arachidonic acid, suggesting the complex mutual relationship of various factors ‐ agonists influencing the function of platelets. This article is protected by copyright. All rights reserved.
    October 30, 2015   doi: 10.1111/1440-1681.12512   open full text
  • Thromboelastography predicts risks of obstetric complication occurrence in (hypo)dysfibrinogenemia patients under non‐pregnant state.
    Jingyi Zhou, Yu Xin, Qiulan Ding, Linlin Jiang, Yaopeng Chen, Jing Dai, Yeling Lu, Xi Wu, Qian Liang, Hongli Wang, Xuefeng Wang.
    Clinical and Experimental Pharmacology and Physiology. October 28, 2015
    Congenital (hypo)dysfibrinogenemia patients may have obstetric complications during their pregnancies. In this study, we aimed to evaluate thromboelastography (TEG) as a potential tool for assessing the tendency of obstetric complications in those patients under non‐pregnant state. A total of 22 female subjects with congenital (hypo)dysfibrinogenemia were recruited. 9 subjects were with histories of obstetric complications and the other 13 subjects had at least one uneventful pregnancy without obstetric complications yet. The detailed clinical investigation and phenotype/genotype detection were carried out, and both kaolin activated TEG and functional fibrinogen TEG (FF‐TEG) were applied in all subjects. Significant difference were identified in all TEG parameters except for R and Angle between these two groups (p<0.05) by covariance analysis. Receiver operating characteristic (ROC) analysis of discrimination between these two groups of patients was performed for TEG parameters. Significantly high odds ratio (OR) of obstetric complications occurrence were demonstrated in K≥3.8 min, MA≤54.2 mm, CI≤‐3 (11.67, 95% CI 1.527‐89.121, p<0.05 in all), and MA‐CFF≤12.1 mm (20.00, 95% CI 1.967‐203.322, p=0.002). Moreover, MA‐CFF had better prognostic performance, with a corresponding area under ROC curve of 0.923 (range 0.815‐1.031, p=0.001). Our study suggests that (hypo)dysfibrinogenemia patients with values out of cut‐off values in TEG assays under non‐pregnant state may have a higher risk of obstetric complications occurrence when they are pregnant. This article is protected by copyright. All rights reserved.
    October 28, 2015   doi: 10.1111/1440-1681.12509   open full text
  • The acid‐sensing ion channels in trigeminal ganglion neurons innervating the orofacial region contribute to orofacial inflammatory pain.
    Hui Fu, Peng Fang, Hai‐Yun Zhou, Jun Zhou, Xiao‐Wei Yu, Ming Ni, Jie‐Yan Zheng, You Jin, Jian‐Guo Chen, Fang Wang, Zhuang‐Li Hu.
    Clinical and Experimental Pharmacology and Physiology. October 28, 2015
    Orofacial pain is a common clinical symptom that is accompanied by tooth pain, migraine, gingivitis, etc. Accumulating evidence suggests that acid‐sensing ion channels (ASICs), especially ASIC3, can profoundly affect the physiological properties of nociception in peripheral sensory neurons. The aim of this study was to examine the contribution of ASICs in trigeminal ganglion (TG) neurons to orofacial inflammatory pain. A western blot (WB), immunofluorescence assay of labelled trigeminal ganglion neurons, orofacial formalin test, cell preparation, and electrophysiological experiments were performed. This study demonstrated that ASIC1, ASIC2a, and ASIC3 are highly expressed in TG neurons innervating the orofacial region of rats. The amplitude of ASIC currents in these neurons increased 119.72% (for ASIC1‐like current) and 230.59% (for ASIC3‐like current) in the formalin‐induced orofacial inflammatory pain model. In addition, WB and immunofluorescence assay demonstrated a significantly augmented expression of ASICs in orofacial TG neurons during orofacial inflammation compared with the control group. The relative protein density of ASIC1, ASIC2a, and ASIC3 also increased 58.82±8.92%, 45.30±11.42%, and 55.32±14.71%, respectively, compared with the control group. Furthermore, pharmacological blockade of ASICs and genetic deletion of ASIC1 attenuated the inflammation response. These findings indicate that peripheral inflammation can induce the upregulation of ASICs in TG neurons, causing orofacial inflammatory pain. Additionally, the specific inhibitor of ASICs may have a significant analgesic effect on orofacial inflammatory pain. This article is protected by copyright. All rights reserved.
    October 28, 2015   doi: 10.1111/1440-1681.12510   open full text
  • Intracoronary electrocardiogram during alcohol septal ablation for hypertrophic obstructive cardiomyopathy predicts myocardial injury size.
    Jing Meng, Xiaolong Qu, Haiyun Huang, Shanwen Zhang, Weibo Zhao, Guoxiang He, Zhiyuan Song, Houyuan Hu.
    Clinical and Experimental Pharmacology and Physiology. October 16, 2015
    Alcohol septal ablation (ASA) has been widely used to treat patients with hypertrophic obstructive cardiomyopathy (HOCM). During the routine ASA procedure, it is difficult to detect the septal injury in real‐time. The aim of the present study was to assess myocardial injury during ASA by recording intracoronary electrocardiogram (IC‐ECG). From 2012 to 2015, 31 HOCM patients were treated with ASA, and IC‐ECG was recorded in 21 patients successfully before and after ethanol injection. The elevation of ST‐segment on IC‐ECG after ethanol injection was expressed as its ratio to the level before injection or the absolute increasing value. Blood samples were collected before and after ASA for measuring changes in cardiac biomarkers. The ratio value of ST‐segment elevation was positively correlated with both the amount of ethanol injected (r = 0.645, P = 0.001) and the myocardial injury size (creatine kinase‐MB area under the curve, AUC of CK‐MB) (r = 0.466, P = 0.017). The absolute increment of ST‐segment was also positively associated with both the amount of ethanol (r = 0.665, P = 0.001) and AUC of CK‐MB (0.685, P = 0.001). However, there was no statistical correlation between the reduction of left ventricular outflow tract gradient and ST‐segment elevation. Additionally no severe ASA procedure‐related complications were observed in our patients. In conclusion, myocardial injury induced by ethanol injection can be assessed immediately by ST‐segment elevation on IC‐ECG. Our study is the first to show that IC‐ECG is a useful method for predicting myocardial injury during ASA in real‐time. This article is protected by copyright. All rights reserved.
    October 16, 2015   doi: 10.1111/1440-1681.12502   open full text
  • The role of maternal nutrition, metabolic function and the placenta in developmental programming of renal dysfunction.
    V F I Richter, J F Briffa, K M Moritz, M E Wlodek, D H Hryciw.
    Clinical and Experimental Pharmacology and Physiology. October 16, 2015
    The intrauterine environment is critical for the development of the fetus. Barker and colleagues were the first to identify that adverse perturbations during fetal development are associated with an increased risk of developing diseases in adulthood, including cardiorenal disease. Specifically for the kidney, perturbations in utero can lead to nephron deficits and renal dysfunction by a number of mechanisms. Altered programming of nephron number is associated with an increased risk of developing kidney disease via glomerular hypertrophy and reduced vasodilative capacity of the renal blood vessels; both of which would contribute to hypertension in adulthood, with the males being more susceptible to disease outcomes. Additionally, alterations in the renin‐angiotensin system (RAS) such as an upregulation or downregulation of specific receptors, depending on the nature of the insult, have also been implicated in the development of renal dysfunction. Sex‐specific differences in the expression of the RAS during late gestation and in the early postnatal environment have also been identified. Extensive research has demonstrated that both uteroplacental insufficiency and maternal malnutrition alter renal development in utero. Equally, exposure to maternal diabetes and maternal obesity during development are also associated with an increased risk of developing renal disease, however the mechanism behind this association is poorly understood. Therefore, identifying the link between an adverse intrauterine environment and the programmed kidney disease risk in adulthood may facilitate the development of strategies to alleviate the epidemics of cardiorenal disease worldwide, in addition to understanding why males are more susceptible to adulthood cardiovascular diseases. This article is protected by copyright. All rights reserved.
    October 16, 2015   doi: 10.1111/1440-1681.12505   open full text
  • The improvement of exercise performance by physical training is related to increased hypothalamic neuronal activation.
    Henrique P. Santiago, Laura H. R. Leite, Paulo Marcelo A. Lima, Gisele V. Rodovalho, Raphael E. Szawka, Cândido C. Coimbra.
    Clinical and Experimental Pharmacology and Physiology. October 16, 2015
    The effects of physical training on hypothalamic activation after exercise and their relationship with heat dissipation were investigated. Following eight weeks of physical training, trained (TR, n=9) and untrained (UN, n=8) Wistar rats were submitted to a regimen of incremental running until fatigue while body and tail temperatures were recorded. After exercise, hypothalamic c‐Fos immunohistochemistry analysis was performed. The workload, body‐heating rate, heat storage and body temperature threshold for cutaneous vasodilation were calculated. Physical training increased the number of c‐Fos immunoreactive neurons in the paraventricular, medial preoptic and median preoptic nucleus by 112%, 90% and 65% (p < 0.01) after exercise, respectively. In these hypothalamic regions, increased neuronal activation was directly associated with the increased workload performed by TR animals (p < 0.01). Moreover, a reduction of 0.6 °C in the body temperature threshold for cutaneous vasodilation was shown by TR animals (p < 0.01). This reduction was possibly responsible for the lower body‐heating rate (0.019 ± 0.002 °C.min−1, TR vs 0.030 ± 0.005 °C, UN, p < 0.05) and the decreased ratio between heat storage and the workload performed by TR animals (18.18 ± 1.65 cal.Kgm−1, TR vs 31.38 ± 5.35 cal.Kgm−1, UN, p < 0.05). The data indicate that physical training enhances hypothalamic neuronal activation during exercise. This enhancement is the central adaptation relating to better physical performance, characterized by a lower ratio of heat stored to workload performed, due to improved heat dissipation. This article is protected by copyright. All rights reserved.
    October 16, 2015   doi: 10.1111/1440-1681.12507   open full text
  • Higher circulating resistin protein and PBMCs resistin mRNA levels are associated with increased prevalence of small dense LDL particles in coronary artery disease patients.
    Jelena Joksić, Miron Sopić, Vesna Spasojević‐Kalimanovska, Tamara Gojković, Aleksandra Zeljković, Jelena Vekić, Kristina Andjelkovic, Dimitra Kalimanovska‐Oštrić, Zorana Jelić‐Ivanović.
    Clinical and Experimental Pharmacology and Physiology. October 15, 2015
    Recent in vitro experiments indicated that human resistin increases the number of lipoprotein particles secreted by the human hepatocytes and also influences their quality, in terms of generating more proatherogenic lipid particles. The aim of this study was to investigate associations of plasma resistin and peripheral blood mononuclear cells (PBMCs) resistin mRNA levels with different prevalence of small, dense low‐density lipoprotein particles (sdLDL) in patients with indications for coronary angiography.This study included 65 patients requiring coronary angiography. There were 41 patients without significant stenosis and 24 patients with significant stenosis in at least one major coronary artery. Circulating resistin was measured by ELISA; PBMC resistin mRNA was determined by real‐time PCR. LDL and HDL subclasses were determined by gradient gel electrophoresis.Plasma resistin (P=0.031) and PBMCs resistin mRNA (P=0.004) were significantly higher in patients with proportion of sdLDL particles ≥50%, compared to the group with relative proportion of sdLDL particles <50%. Plasma resistin correlated positively with creatinine (r=0.456, P<0.001) and resistin mRNA (r=0.298, P=0.014) but negatively with BMI (r=‐0.254, P=0.034) and TC (r=‐0.286, P=0.021). Multiple linear regression analysis revealed LDL particle diameter as the only independent predictor of resistin mRNA (R2=0.258; adjR2=0.190).A significant association between resistin, both PBMCs mRNA and plasma protein, and the relative proportion of sdLDL particles in the circulation of CAD patients has been established, which implies that increased gene expression of resistin in PBMCs and higher resistin concentration in plasma are related to pro‐atherogenicLDL particle phenotype. This article is protected by copyright. All rights reserved.
    October 15, 2015   doi: 10.1111/1440-1681.12503   open full text
  • Depotentiation of intact rat cardiac muscle unmasks an Epac‐dependent increase in myofilament Ca2+ sensitivity.
    Sarbjot Kaur, Cherrie H.T. Kong, Mark B. Cannell, Marie‐Louise Ward.
    Clinical and Experimental Pharmacology and Physiology. October 15, 2015
    Recently, a family of guanine nucleotide exchange factors have been identified in many cell types as important effectors of cyclic adenosine 3′,5′‐monophospahte (cAMP) signalling that is independent of protein kinase A (PKA). In the heart, investigation of exchange protein directly activated by cAMP (Epac) has yielded conflicting results. Since cAMP is an important regulator of cardiac contractility, our aim was to examine whether Epac activation modulates excitation‐contraction coupling in ventricular preparations from rat hearts. We used 8‐(4‐chlorophenylthio)‐2′‐O‐methyladenosine‐3′, 5′‐cyclic monophosphate (cpTOME), an analogue of cAMP that activates Epac, but not PKA. In isolated myocytes, cpTOME increased Ca2+ spark frequency from ~ 7 to 32 per 100 μm3 s (n = 10), P = 0.05 with a reduction in the peak amplitude of the sparks. Simultaneous measurements of intracellular Ca2+ and isometric force in multicellular trabeculae (n = 7, 1.5 mM [Ca2+]o) revealed no effect of Epac activation on either the amplitude of Ca2+ transients (Control: 0.7 ± 0.1 versus cpTOME: 0.7 ± 0.1 340/380 fura‐2 ratio, P = 0.35) or on peak stress (Control: 24 ± 5 versus cpTOME: 23 ± 5 mN mm−2, P = 0.20). However, an effect of Epac in trabeculae was unmasked by lowering extracellular [Ca2+]o. In these depotentiated trabeculae, activation of the Epac pathway increased myofilament Ca2+ sensitivity, an effect that was blocked by addition of KN‐93, a Ca2+/calmodulin‐dependent protein kinase II (CaMK‐II) inhibitor. This study suggests that Epac activation may be a useful therapeutic target to increase the strength of contraction during low inotropic states. This article is protected by copyright. All rights reserved.
    October 15, 2015   doi: 10.1111/1440-1681.12504   open full text
  • Sex differences in the mechano‐energetic effects of genistein on stunned rat and guinea pig hearts.
    Germán A. Colareda, María I. Ragone, Alicia E. Consolini.
    Clinical and Experimental Pharmacology and Physiology. October 09, 2015
    Although the phytoestrogen genistein (Gen) is considered protective in cardiovascular diseases, its direct effects on stunned hearts after transient ischemia‐reperfusion (I/R) are unknown. Here we studied the effects of 20 μM Gen on the mechano‐calorimetric behavior during I/R of rat and guinea pig hearts to evaluate the energetics of Ca2+ homeostasis. Isolated beating hearts were perfused with control Krebs solution inside a calorimeter with or without perfusion of Gen before a transient period of I/R. Left ventricular pressure development (P) and total heat rate (Ht) were continuously measured. At 37 °C, Gen did not change post‐ischemic contractile recovery (PICR), but it increased the relaxation rate. However, PICR was reduced in hearts of male rats and guinea pigs at 30 °C. Total muscle economy (P/Ht) showed the same behavior as P at each temperature. Inhibition of phosphatases with orthovanadate during Gen perfusion prevented a decrease in PICR in male rat hearts, suggesting that this effect is due to tyrosine kinase inhibition. Reperfusing ischemic hearts with 10 mM caffeine‐36 mM Na+‐Krebs induced contracture dependent on the sarcoreticular Ca2+ content. Contracture relaxation depends on mitochondrial Ca2+ uptake and Gen reduced the relaxation rate. Moreover, Gen prevented the increase in Rhod‐2 fluorescence (free [Ca2+]m) of rat cardiomyocytes. In guinea pig hearts, Gen maintained ischemic preconditioning, but was reduced by 5‐hydroxydecanoate, suggesting the participation of mitochondrial ATP‐dependent K channels. Results suggest that Gen acts on several mechanisms that regulate myocardial calcium homeostasis and energetics during I/R, which differ in a temperature‐ and sex‐dependent manner. This article is protected by copyright. All rights reserved.
    October 09, 2015   doi: 10.1111/1440-1681.12500   open full text
  • Slowed Atrial and Atrioventricular Conduction and Depressed HRV in a Murine Model of Hypertrophic Cardiomyopathy.
    Wei‐Wen Lim, Mathias Baumert, Melissa Neo, Pawel Kuklik, Anand N Ganesan, Dennis H Lau, Tatiana Tsoutsman, Christopher Semsarian, Prashanthan Sanders, David A Saint.
    Clinical and Experimental Pharmacology and Physiology. October 07, 2015
    Hypertrophic cardiomyopathy (HCM) is a common heritable cardiac disorder with diverse clinical outcomes including sudden death, heart failure, and stroke. Depressed heart rate variability (HRV), a measure of cardiac autonomic regulation, has been shown to predict mortality in patients with cardiovascular disease. Cardiac autonomic remodelling in animal models of HCM are not well characterised. We studied the Gly203Ser cardiac troponin‐I transgenic (TG) male mice previously demonstrated to develop hallmarks of HCM by age 21 weeks. 33 mice aged 30 and 50 weeks underwent continuous electrocardiogram (ECG) recording for 30 minutes under anesthesia. TG mice demonstrated prolonged P‐wave duration (p<0.001) and PR intervals (p<0.001) compared to controls. Additionally, TG mice demonstrated depressed standard deviation of RR intervals (SDRR; p<0.01), coefficient of variation of RR intervals (CVRR; p<0.001) and standard deviation of heart rate (SDHR; p<0.001) compared to controls. Additionally, total power was significantly reduced in TG mice (p<0.05). No significant age‐related difference in either strain was observed in ECG or HRV parameters. Mice with HCM developed slowed atrial and atrioventricular conduction and depressed HRV. These changes were conserved with increasing age. This finding may be indicative of atrial and ventricular hypertrophy or dysfunction, and perhaps an indication of worse clinical outcome in heart failure progression in HCM patients. This article is protected by copyright. All rights reserved.
    October 07, 2015   doi: 10.1111/1440-1681.12498   open full text
  • Increased Ratio of Neutrophil Elastase to α1‐antitrypsin Is Closely Associated with Liver Inflammation in Patients with Nonalcoholic Steatohepatitis.
    Shufei Zang, Xiaojie Ma, Zhenjie Zhuang, Jing Liu, Dongxue Bian, Yunhao Xun, Qiuling Zhang, Falin Zhao, Wenjun Yang, Juan Liu, Yan Luo, Yinlan Liu, Bei Ye, Dewei Ye, Junping Shi.
    Clinical and Experimental Pharmacology and Physiology. October 07, 2015
    An imbalance between neutrophil elastase (NE) and its inhibitor α1‐antitrypsin (A1AT) is known to contribute to the development of obesity‐related inflammation. This study aimed to investigate the role of the NE‐A1AT system in the histological progression of non‐alcoholic fatty liver disease (NAFLD), and to evaluate the ability of it to predict nonalcoholic steatohepatitis (NASH). A total of 252 adults (NAFLD group, n = 202; Healthy group, n = 50) were recruited. Clinical biochemical characteristics, NE and A1AT concentrations were measured in all subjects. Among the NAFLD group, 86 patients had previously undergone liver biopsy and information on histological characteristics was consequently available. The area under receiver operating characteristic curve (AUC) was used to determine the predictive accuracy of the NE‐A1AT system for NASH. NAFLD patients had an elevated serum NE concentration and a reduced A1AT level with consequent NE/A1AT imbalance. NE increased in the early stage of steatosis, preceded the decline in A1AT, dating from the onset of NASH (NAS 3‐4), and subsequent NE/A1AT increased in the presence of NASH. Nonetheless, this increase began to resolve as the disease state progressed to advanced fibrosis. A1AT had a sensitivity (SEN) of 83.8% and a specificity (SP) of 83.3% with the optimal cut‐off of ‐1459.43, NE/A1AT had a SEN of 88.8% and a SP of 83.3% with cut‐off of 0.363 to predict NASH. An increased NE: A1AT ratio is closely associated with liver Inflammation in patients with NASH and could serve as a novel marker to predict NASH in humans. This article is protected by copyright. All rights reserved.
    October 07, 2015   doi: 10.1111/1440-1681.12499   open full text
  • Role of cyclooxygenae‐1 and ‐2 in endothelium‐dependent contraction of atherosclerotic mouse abdominal aortas.
    Shasha Li, Bin Liu, Wenhong Luo, Yingzhan Zhang, Hui Li, Dongyang Huang, Yingbi Zhou.
    Clinical and Experimental Pharmacology and Physiology. October 07, 2015
    Objective To determine the role of cyclooxygenase (COX)‐1 or ‐2 in endothelium‐dependent contraction under atherosclerotic conditions. Methods Atherosclerosis was induced in apoE knockout (apoE‐/‐) mice and those with COX‐1‐/‐ (apoE‐/‐‐COX‐1‐/‐) by feeding with high fat and cholesterol food. Aortas (abdominal or the whole section) were isolated for functional and/or biochemical analyses. Results As in non‐atherosclerotic conditions, the muscarinic receptor agonist acetylcholine (ACh) evoked an endothelium‐dependent, COX‐mediated contraction following NO synthase (NOS) inhibition in abdominal aortic rings from atherosclerotic apoE‐/‐ mice. Interestingly, COX‐1 inhibition not only abolished such a contraction in rings showing normal appearance, but also diminished that in rings with plaques. Accordingly, only a minor contraction (<30% that of apoE‐/‐ counterparts) was evoked by ACh (following NOS inhibition) in abdominal aortic rings of atherosclerotic apoE‐/‐‐COX‐1‐/‐ mice with plaques, and none was evoked in those showing normal appearance. Also, the contraction evoked by ACh in apoE‐/‐‐COX‐1‐/‐ abdominal aortic rings with plaques was abolished by non‐selective COX inhibition, thromboxane‐prostanoid (TP) receptor antagonism, or endothelial denudation. Moreover, we noted that ACh evoked a predominant production of the prostacyclin (PGI2, which mediates abdominal aortic contraction via TP receptors in mice) metabolite 6‐keto‐PGF1α, which was again sensitive to COX‐1 inhibition or COX‐1‐/‐. Conclusion In atherosclerotic mouse abdominal aortas, COX‐1 can still be the major isoform mediating endothelium‐dependent contraction, which probably results largely from PGI2 synthesis as in non‐atherosclerotic conditions. In contrast, COX‐2 may have only a minor role in such response limited to areas of plaques under the same pathological condition. This article is protected by copyright. All rights reserved.
    October 07, 2015   doi: 10.1111/1440-1681.12501   open full text
  • CCN2 and CCN5 exerts opposing effect on fibroblast proliferation and transdifferentiation induced by TGF‐β.
    Honghai Xu, Peng Li, Mengting Liu, Cong Liu, Zhengming Sun, Xiong Guo, Yuelin Zhang.
    Clinical and Experimental Pharmacology and Physiology. October 06, 2015
    Epidural fibrosis might occur after lumbar discectomy and contributes to failed back syndrome. Transforming growth factor (TGF)‐β has been reported to influence multiple organ fibrosis, in which connective tissue growth factor/cysteine‐rich 61/nephroblastoma overexpressed 2 (CCN2) and CCN5 are involved. However, the effect of CCN2 and CCN5 on TGF‐β induced fibrosis has not yet been elucidated. This study reports that CCN2 and CCN5 play opposing roles in cell proliferation and transdifferentiation of human skin fibroblasts or rabbit epidural scar‐derived fibroblasts exposed to TGF‐β. We observed that TGF‐β1 induced fibroblasts proliferation and differentiation in a dose‐dependent manner (from 0 μg/L to 20 μg/L). Meanwhile, CCN2 expression is up‐regulated while CCN5 expression is inhibited by TGF‐β1 exposure. Furthermore, it is demonstrated that CCN2 overexpression leads to promoted proliferation and elevated collagen and α‐smooth muscle actin (α‐SMA) expression, which are inhibited by CCN5 overexpression. Moreover, it is shown that the cysteine knot (CT) domain, present in CCN2 but absent in CCN5, plays an essential part in fibroblast proliferation and differentiation. Additionally, enhanced TGF‐β and CCN2 expression but decreased CCN5 expression is found in rabbit epidural scar‐derived fibroblasts. Overall, the results show the opposing effects of CCN2 and CCN5 on fibroblast proliferation and transdifferentiation induced by TGF‐β.
    October 06, 2015   doi: 10.1111/1440-1681.12470   open full text
  • The length‐dependent activation of contraction is equally impaired in impuberal male and female rats in monocrotaline‐induced right ventricular failure.
    Oleg Lookin, Alexander Balakin, Daniil Kuznetsov, Yuri Protsenko.
    Clinical and Experimental Pharmacology and Physiology. October 06, 2015
    The length‐dependent activation of contraction is attenuated in the failing myocardium of adult male rats. This pathological change is not seen in adult female rats, possibly because of a protective effect of sex hormones. The present study evaluated length‐dependent changes in isometric twitch, Ca2+ transient (CaT) and action potential (AP) in the right ventricular myocardium of impuberal healthy male and female rats (control) and in rats treated with a single injection of 50 mg/kg monocrotaline (MCT). Compared with sex‐matched control rats, MCT‐treated male and female rats exhibited increased right ventricular weight (134% and 142% of control, respectively), decreased left ventricular weight (72% and 79%), twitch attenuation (48.8 ± 2.7% and 57.5 ± 1.2%) and prolongation (125 ± 3% and 127 ± 2%), CaT attenuation (37.8 ± 0.4% and 39.1 ± 1.1%) and prolongation (114 ± 1% and 116 ± 1%) and AP prolongation at 90% repolarization (195 ± 2% and 203 ± 1%). The MCT‐treated male rats exhibited a 50% lower integral magnitude and an approximately 25% larger time‐to‐peak ‘bump’ compared with control male rats. These parameters in MCT‐treated female rats tended to show similar changes to those seen in the control female rats, with no significant difference between the two groups. In all groups, integral magnitude and time‐to‐peak ‘bump’ increased with length. In conclusion, the length‐dependent activation of contraction was equally blunted in the failing right ventricular myocardium of impuberal male and female rats. This was related to changes in CaT and AP, which were similar between male and female rats. Therefore, puberty is necessary for manifestation of the protective effects of sex hormones on this remodelling.
    October 06, 2015   doi: 10.1111/1440-1681.12471   open full text
  • Asiatic acid alleviates cardiovascular remodelling in rats with L‐NAME‐induced hypertension.
    Sarawoot Bunbupha, Parichat Prachaney, Upa Kukongviriyapan, Veerapol Kukongviriyapan, Jariya Umka Welbat, Poungrat Pakdeechote.
    Clinical and Experimental Pharmacology and Physiology. October 06, 2015
    A previous study demonstrated the antihypertensive effect of asiatic acid. The current study investigates the effect of asiatic acid on cardiovascular remodelling and possible mechanisms involved in Nω‐nitro‐L‐arginine methyl ester hydrochloride (L‐NAME)‐induced hypertensive rats. Male Sprague–Dawley rats were treated with L‐NAME (40 mg/kg per day) for 3 weeks in order to induce hypertension. Hypertensive rats were administered asiatic acid (20 mg/kg per day) or vehicle for a further 2 weeks. It was found that hypertensive rats showed high systolic blood pressure, left ventricular (LV) hypertrophy, increases in LV fibrosis, aortic wall thickness and aortic collagen deposition (P < 0.05). Moreover, decreased plasma nitrate and nitrite (NOx) and increased plasma tumor necrosis factor alpha (TNF‐α) were observed in hypertensive rats (P < 0.05). This was consistent with downregulation of endothelial nitric oxide synthase (eNOS) expression and upregulation of inducible nitric oxide synthase (iNOS) expression in heart and aortic tissues (P < 0.05). Levels of malondialdehyde (MDA) in plasma, aortic and heart tissues were significantly increased in hypertensive rats (P < 0.05). Asiatic acid markedly reduced blood pressure, alleviated cardiovascular remodelling, and restored plasma NOx and TNF‐α as well as eNOS/iNOS expression in heart and aortic tissues (P < 0.05). Additionally, there was a significant reduction of MDA levels in the tissues of treated hypertensive rats. In conclusion, this study demonstrates the therapeutic effects of asiatic acid on blood pressure and cardiovascular remodelling, which is possibly related to the restoration of eNOS/iNOS expression, and the resulting anti‐inflammatory and antioxidant activities.
    October 06, 2015   doi: 10.1111/1440-1681.12472   open full text
  • Angiotensin and mineralocorticoid receptor antagonism attenuates cardiac oxidative stress in angiotensin II‐infused rats.
    Jacqueline N Minas, Max A Thorwald, Debra Conte, Jose‐Pablo Vázquez‐Medina, Akira Nishiyama, Rudy M Ortiz.
    Clinical and Experimental Pharmacology and Physiology. October 06, 2015
    Angiotensin II (Ang II) and aldosterone contribute to hypertension, oxidative stress and cardiovascular damage, but the contributions of aldosterone during Ang II‐dependent hypertension are not well defined because of the difficulty to assess each independently. To test the hypothesis that during Ang II infusion, oxidative and nitrosative damage is mediated through both the mineralocorticoid receptor (MR) and angiotensin type 1 receptor (AT1), five groups of Sprague–Dawley rats were studied: (i) control; (ii) Ang II infused (80 ng/min × 28 days); (iii) Ang II + AT1 receptor blocker (ARB; 10 mg losartan/kg per day × 21 days); (iv) Ang II + mineralocorticoid receptor (MR) antagonist (Epl; 100 mg eplerenone/day × 21 days); and (v) Ang II + ARB + Epl (Combo; × 21 days). Both ARB and combination treatments completely alleviated the Ang II‐induced hypertension, whereas eplerenone treatment only prolonged the onset of the hypertension. Eplerenone treatment exacerbated the Ang II‐mediated increase in plasma and heart aldosterone 2.3‐ and 1.8‐fold, respectively, while ARB treatment reduced both. Chronic MR blockade was sufficient to ameliorate the AT1‐mediated increase in oxidative damage. All treatments normalized protein oxidation (nitrotyrosine) levels; however, only ARB and Combo treatments completely reduced lipid peroxidation (4‐hydroxynonenal) to control levels. Collectively, these data suggest that receptor signalling, and not the elevated arterial blood pressure, is the principal culprit in the oxidative stress‐associated cardiovascular damage in Ang II‐dependent hypertension.
    October 06, 2015   doi: 10.1111/1440-1681.12473   open full text
  • Immunomodulation by splenectomy or by FTY720 protects the heart against ischemia reperfusion injury.
    D Goltz, S Huss, E Ramadori, R Büttner, L Diehl, R Meyer.
    Clinical and Experimental Pharmacology and Physiology. October 06, 2015
    The pathogenesis of myocardial ischemia‐reperfusion injury (MI/R) involves an inflammatory response in the myocardium undergoing reperfusion. Modulation of this response by splenectomy constitutes an option to protect the heart from MI/R. To mimic the effect of splenectomy in a pharmacological approach, the sphingosine‐1‐phosphate agonist FTY720 was applied at the onset of reperfusion. In a closed chest model of MI/R, infarct size was assessed by triphenyltetrazolium chloride staining after 1 h of ischemia and 24 h of reperfusion, and by Masson trichrome staining 21 days after reperfusion in splenectomised mice, mice post‐conditioned with FTY720 IP (1 mg/kg), and controls. In addition, hemodynamic parameters were recorded after 24 h and 21 days by catheterization. Infarct size, and immune cell invasion of phagocytic monocytes investigated by FACS after 24 h of reperfusion were significantly reduced by both splenectomy, and FTY720 treatment. Evaluation after 21 days of reperfusion revealed that FTY720 treated animals had an improved hemodynamic outcome compared to placebo treated as well as splenectomised animals. FTY720 treatment reduced cell injury as effectively as splenectomy by lowering the number of phagocytic monocytes invading the myocardium and ameliorated hemodynamic outcome within the first 21 days.
    October 06, 2015   doi: 10.1111/1440-1681.12465   open full text
  • Multipotent bone marrow stromal cell therapy promotes endogenous cell proliferation following ischemic stroke.
    Gila Pirzad Jahromi, Alireza Shabanzadeh Pirsaraei, Seyed Shahabeddin Sadr, Golamreza Kaka, Mahvash Jafari, Sadegh Seidi, Jason Charish.
    Clinical and Experimental Pharmacology and Physiology. October 06, 2015
    Despite extensive research over the years, there still exists some debate as to what constitutes the optimal therapeutic strategy to promote recovery following stroke. Due to the complexity of injured brain pathophysiology, treatment approaches should ideally address numerous factors, ultimately aiming to promote tissue protection, axonal regrowth and functional recovery. This study extends the understanding of the effects of bone marrow stromal cell (BMSC) treatment following experimentally induced ischemic stroke in rats. Focal ischemic brain injury was experimentally induced in rats by placing a preformed clot into the middle cerebral artery. Animals were injected intravenously with BMSCs at 24 h after stroke and were killed 7 days post injury. When administered BMSCs following stroke, the neurological outcome was significantly improved relative to controls. There was an increase in the number of BMSCs labelled with BrdU present in the injured hemisphere of the brain compared to the non‐injured side. Furthermore, administration of BMSCs also led to increases in astrocytosis, vascularization and endogenous proliferation. These findings provide insight into the mechanisms of action of BMSC treatment and further argue for the therapeutic potential of BMSCs as an effective treatment following cerebral stroke.
    October 06, 2015   doi: 10.1111/1440-1681.12466   open full text
  • Fibroblast growth factor 23 is associated with the presence of coronary artery disease and the number of stenotic vessels.
    Xiang Hu, Xiaojing Ma, Xiaoping Pan, Yaping Hao, Yuqi Luo, Zhigang Lu, Yuqian Bao, Weiping Jia.
    Clinical and Experimental Pharmacology and Physiology. October 06, 2015
    Fibroblast growth factor 23 (FGF23) has been reported to be involved in cardiovascular disease. The aim of this study was to investigate the association between FGF23 and the presence of coronary artery disease (CAD), as well as the number of stenotic vessels. A total of 254 eligible participants (167 men and 87 postmenopausal women) were enrolled in this study. Coronary angiography was used for diagnosis of CAD. Serum intact FGF23 levels were determined by a two‐sided sandwich enzyme‐linked immunosorbent assay. The median serum FGF23 levels of the entire study population were 39.9 (33.1–47.5) pg/mL. Serum FGF23 levels were higher in subjects with one‐vessel disease than those without CAD (P < 0.05), which further increased significantly in the subjects with multi‐vessel disease (P < 0.05). Serum FGF23 levels increased with cumulative number of stenotic vessels (P for trend < 0.001). Multiple stepwise regression analysis revealed estimated glomerular filtration rate (standardized β = −0.298; P < 0.001) and body mass index (standardized β = 0.132; P = 0.049) were independent factors correlated with FGF23. Multivariate logistic regression analysis showed that FGF23 was positively and independently associated with the presence of CAD (odds ratio = 1.058, 95% confidence interval = 1.025–1.092; P = 0.001). Additionally, FGF23 was also correlated with multi‐vessel disease significantly (odds ratio = 1.034, 95% confidence interval = 1.007–1.062; P = 0.013). In conclusion, serum FGF23 levels exhibit positive and independent association with the presence of CAD and increase with the cumulative number of stenotic vessels.
    October 06, 2015   doi: 10.1111/1440-1681.12467   open full text
  • The expression of the imprinted gene pleckstrin homology‐like domain family A member 2 in placental tissues of preeclampsia and its effects on the proliferation, migration and invasion of trophoblast cells JEG‐3.
    Feng Jin, Chong Qiao, Nannan Luan, Tao Shang.
    Clinical and Experimental Pharmacology and Physiology. October 06, 2015
    Preeclampsia (PE) is one of the most common hypertensive disorders and is a leading cause of morbidity and mortality for pregnant women and perinatal babies. Additionally, pleckstrin homology‐like domain family A member 2 (PHLDA2) is associated with placental dysfunction. However, the effect of PHLDA2 on trophoblast cell proliferation, migration and invasion has not been investigated. In this study, 15 PE patients and 15 normal pregnant women were recruited and clinical characteristics were summarized. Pleckstrin homology‐like domain family A member 2 levels in placental tissues were examined using real‐time PCR and western blot. Overexpression plasmid and PHLDA2 siRNA was introduced into JEG‐3 cells, respectively. Cell proliferation was measured using MTT assay and flow cytometry. Cell migration and invasion capacities were assessed by wound healing and Transwell assays. It was found that PE patients collectively presented proteinuria, elevated systolic blood pressure (SBP) and diastolic blood pressure (DBP), and lower gestational ages and birth weights. Pleckstrin homology‐like domain family A member 2 levels in the preeclamptic placenta were significantly upregulated. Pleckstrin homology‐like domain family A member 2 overexpression significantly arrested cells in the G0/G1 phase, inhibited cell proliferation and suppressed the migration and invasion of JEG‐3 cells. Pleckstrin homology‐like domain family A member 2 knockdown significantly blocked the cells in the S phase of the cell cycle. Knockdown of PHLDA2 alleviated the inhibition on the migration and invasion of trophoblast cells JEG‐3. These findings illustrate that PHLDA2 may participate in PE pathogenesis and indicate its potential application in the early diagnosis of PE.
    October 06, 2015   doi: 10.1111/1440-1681.12468   open full text
  • Anti‐inflammatory Role of Obestatin in Autoimmune myocarditis.
    Ozge Pamukcu, Ali Baykan, Latife Cakir Bayram, Figen Narin, Nazmi Cetin, Nazmi Narin, Mustafa Argun, Abdullah Ozyurt, Kazim Uzum.
    Clinical and Experimental Pharmacology and Physiology. October 01, 2015
    Obestatin is a popular endogeneous peptide, known to have an autoimmune regulatory effect on energy metabolism and the gastrointestinal system. Studies regarding the anti‐inflammatory effects of obestatin are scarce. In this study, we aimed to show the anti‐inflammatory effect of obestatin in an experimental model of autoimmune myocarditis in rats. Experimental autoimmune myocarditis was induced in Lewis rats by immunization with subcutaneous administration of porcine cardiac myosin, twice at 7‐day intervals. Intraperitoneal pretreatment with obestatin (50μg/kg) was started before the induction of myocarditis and continued for 3 weeks. The severity of myocarditis was evidenced by clinical, echocardiographic and histological findings. In addition, by‐products of neutrophil activation, lipid peroxidation, inflammatory and anti‐inflammatory cytokines were measured in serum. Obestatin significantly ameliorated the clinical and histopathological severity of autoimmune myocarditis. Therapeutic effects of obestatin in myocarditis were associated with reduced lipid peroxidation, suppression of polymorphonuclear leukocyte (PMNL) infiltration and enhancement of glutathione synthesis, inhibition of serum inflammatory and activation of anti‐inflammatory cytokines. Histopathologically, the left ventricle was significantly dilated, and its wall thickened, along with widespread lymphocytic and histocytic infiltration. The myocardium was severely infiltrated with relatively large mononuclear cells. These histopathological changes were observed in lesser degrees in obestatin‐treated rats. This study demonstrated a novel anti‐inflammatory effect of obestatin in an experimental model of autoimmune myocarditis. Consequently, obestatin administration may represent a promising therapeutic approach for myocarditis and dilated cardiomyopathy in the future. This article is protected by copyright. All rights reserved.
    October 01, 2015   doi: 10.1111/1440-1681.12497   open full text
  • Low‐dose adjunctive cilostazol in patients with complex lesions undergoing percutaneous coronary intervention.
    Xin‐Tian Zheng, Kang‐Yin Chen, Tong Liu, Ling‐Xia Xu, Jing‐Jin Che, Seung‐Woon Rha, Guang‐Ping Li.
    Clinical and Experimental Pharmacology and Physiology. September 28, 2015
    Patients with complex coronary lesions undergoing percutaneous coronary intervention (PCI) have more major adverse cardiac events (MACE) than do those with simpler one. Therefore, intensive antiplatelet therapy might be needed in these patients. A total of 127 patients with complex lesion undergoing PCI in the Second Hospital of Tianjin Medical University from October 2012 to April 2014 were randomized to receive: dual (aspirin plus clopidogrel, DAPT, n=66) or triple antiplatelet therapy (aspirin plus clopidogrel plus cilostazol, TAPT, n=61). Patients in TAPT group received low‐dose cilostazol (100mg loading, followed with 50mg BID) for 3‐6 months. The primary endpoint was composite MACE. The complex coronary target lesions were defined as at least one of the followings: left main disease, severe 3‐vessel disease, chronic total occlusion lesions, true bifurcation lesion, ostial lesions, severe calcified lesions and highly thrombotic lesions. The two groups had similar baseline clinical and angiographic characteristics. One‐year clinical outcomes showed that TAPT group had significantly lower incidences of myocardial infarction (1.6% vs 13.6%, P=0.018) and MACE (1.6% vs 16.7%, P=0.004) than DAPT group. DAPT group had 2 cases of stent thrombosis, while TAPT group didn't. Furthermore, adjunctive low‐dose cilostazol didn't significantly increase the incidence of bleeding events (26.2% vs 19.7%, P=0.381) regardless of major (4.9% vs 4.5%, P=0.921) or minor (21.3% vs 15.2%, P=0.368) bleeding events. In conclusion, low‐dose adjunctive cilostazol seems superior to dual antiplatelet therapy in reducing recurrent ischemic events in patients with complex coronary lesions and two groups have similar incidence of bleeding events. This article is protected by copyright. All rights reserved.
    September 28, 2015   doi: 10.1111/1440-1681.12495   open full text
  • HDAC4 stabilizes SIRT1 via Sumoylation SIRT1 to delay cellular senescence.
    Xiaolin Han, Jing Niu, Yang Zhao, Qingsheng Kong, Tanjun Tong, Limin Han.
    Clinical and Experimental Pharmacology and Physiology. September 28, 2015
    The NAD‐dependent protein deacetylase Sir2 (silent information regulator 2) regulates cellular lifespan in several organisms. Histone deacetylase 4 (HDAC4) belongs to the class IIa group of histone deacetylases; this class of HDACs is composed of proteins that are important regulators of gene expression that control pleiotropic cellular functions. However, the role of HDAC4 in cellular senescence is still unknown. Here, we show that the expression patterns of HDAC4 and SIRT1 (Sirtuin 1, the mammalian homolog of Sir2) are positively correlated during cellular senescence. Moreover, the overexpression of HDAC4 delays senescence, whereas the knockdown of HDAC4 leads to premature senescence in human fibroblasts. Furthermore, we demonstrate that HDAC4 increases endogenous SIRT1 expression by enhancing its sumoylation modification levels, thereby stabilizing its protein levels. Therefore, our study provides a new molecular mechanism for the regulation of cellular senescence. This article is protected by copyright. All rights reserved.
    September 28, 2015   doi: 10.1111/1440-1681.12496   open full text
  • Increased Sphingosine 1‐phosphate mediates inflammation and fibrosis in tubular injury in diabetic nephropathy.
    Dania Yaghobian, Anthony S. Don, Sarina Yaghobian, Xinming Chen, Sonia Saad, Carol Pollock.
    Clinical and Experimental Pharmacology and Physiology. September 28, 2015
    Hyperglycemia induces all isoforms of transforming growth factor β (TGFβ), which in turn play key roles in inflammation and fibrosis that characterize diabetic nephropathy. Sphingosine 1‐phosphate (S1P) is a signaling sphingolipid, derived from sphingosine by the action of sphingosine kinase (SK). S1P mediates many biological processes, which mimic TGFβ signaling. To determine the role of SK1 and S1P in inducing fibrosis and inflammation, and the interaction with TGFβ‐1, 2 and 3 signalling in diabetic nephropathy, human proximal tubular cells (HK2 cells) were exposed to normal (5mM) or high (30mM) glucose or TGFβ‐1, ‐2, ‐3 +/‐ an SK inhibitor (SKI‐II) or SK1 siRNA. Control and diabetic wild type (WT) and SK1‐/‐ mice were studied. Fibrotic and inflammatory markers, and relevant downstream signaling pathways were assessed. SK1 mRNA and protein expression was increased in HK2 cells exposed to high glucose or TGFβ1,‐2,‐3. All TGFβ isoforms induced fibronectin, collagen IV and macrophage chemoattractant protein 1 (MCP1), which were reversed by both SKI‐II and SK1 siRNA. Exposure to S1P increased phospho‐p44/42 expression, AP‐1 binding and NFkB phosphorylation. WT diabetic mice exhibited increased renal cortical S1P, fibronectin, collagen IV and MCP1 mRNA and protein expression compared to SK1‐/‐ diabetic mice. In summary, this study demonstrates that inhibiting the formation of S1P reduces tubulointerstitial renal inflammation and fibrosis in diabetic nephropathy. This article is protected by copyright. All rights reserved.
    September 28, 2015   doi: 10.1111/1440-1681.12494   open full text
  • PI3K/Akt/mTOR Pathway Dual Inhibitor BEZ235 Suppresses the Stemness of Colon Cancer Stem Cells.
    Jiezhong Chen, Renfu Shao, Feng Li, Michael Monteiro, Jun‐Ping Liu, Zhi Ping Xu, Wenyi Gu.
    Clinical and Experimental Pharmacology and Physiology. September 24, 2015
    Colon cancer is one of the most common cancers worldwide with high mortality. A major issue in colon cancer treatment is drug‐resistance and metastasis that have been ascribed to the cancer stem cells. In this study, we isolated colon cancer stem cells through sphere culture and verified with the cancer stem cell markers CD133, CD44, and CD24. We demonstrated that the PI3K/Akt/mTOR signalling pathway was highly activated in the colon cancer stem cells and that inhibition of the PI3K/Akt/mTOR pathway by the inhibitor BEZ235 suppressed the colon cancer stem cell proliferation with reduced stemness indicated by CD133 and Lgr5 expressions. Treatment with insulin as a known activator of the PI3K/Akt pathway increased CD133 expression and decreased the effects of BEZ235 on colon cancer proliferation and survival. Our data collectively suggest that the PI3K/Akt/mTOR pathway underpins the stemness of colon cancer stem cells and BEZ235 is a good drug candidate potentially for treatment of colon cancer drug resistance and metastasis. This article is protected by copyright. All rights reserved.
    September 24, 2015   doi: 10.1111/1440-1681.12493   open full text
  • Antiarrhythmic activity in occlusion‐reperfusion model of 1‐(1H‐indol‐4‐yloxy)‐3‐{[2‐(2‐methoxyphenoxy)ethyl]amino} propan‐2‐ol and its enantiomers.
    Marek Bednarski, Monika Otto, Magdalena Dudek, Agata Siwek, Małgorzata Zygmunt, Joanna Knutelska, Leszek Nowiński, Grażyna Groszek, Jacek Sapa.
    Clinical and Experimental Pharmacology and Physiology. September 19, 2015
    Acute myocardial infarction (AMI) is a leading cause of mortality and morbidity worldwide, especially in developed countries. The most serious problem after myocardial infarction is reperfusion injury that is manifest as functional impairment, arrhythmia, and accelerated progression of cell death in certain critically injured myocytes. Subsequently the infarcted myocardium develops features of necrosis and reactive inflammation. To reduce lethal reperfusion injury in patient with AMI we use antioxidants, anti‐inflammatory agents, adenosine, opioids, metabolic modulators (glucose, insulin, and potassium, nicorandil and agents which reduce intracellular Ca2+ overload and inhibit Na+‐H+ exchange). In our study a novel compound (9) 1‐(1h‐indol‐4‐yloxy)‐3‐{[2‐(2‐methoxyphenoxy) ethyl]amino}propan‐2‐ol and its enantiomers were examined in arrhythmia associated with coronary artery occlusion and reperfusion rat model. For test compounds were also determined antioxidant properties using MDA and FRAP tests. In summary, the tested compounds, especially the S enantiomer has a strong antiarrhythmic activity in a model of occlusion and reperfusion of the left coronary artery which is probably related to their adrenolytic action. In contrast, to carvedilol none of the test compound reduced the lipid peroxidation but increase ferric reducing antioxidant power. In the antioxidant effect, there was no difference between the optical forms of compound 9. This article is protected by copyright. All rights reserved.
    September 19, 2015   doi: 10.1111/1440-1681.12491   open full text
  • Oxygen‐glucose Deprivation Preconditioning Protects Neurons against Oxygen‐glucose Deprivation/reperfusion Induced Injury via Bone Morphogenetic Protein‐7 Mediated ERK, p38 and Smad Signaling Pathways.
    Junhong Guan, Shaonan Du, Tao Lv, Shengtao Qu, Qiang Fu, Ye Yuan.
    Clinical and Experimental Pharmacology and Physiology. September 19, 2015
    Bone morphogenetic protein (BMP)‐7 mediated neuroprotective effect of cerebral ischemic preconditioning (IPC) has been studied in ischemic animal model, but the underlying cellular mechanisms have not been clearly clarified. In this study, primary cortical neurons and SH‐SY5Y cell line were used to investigate the role of BMP‐7 and its downstream signals in the neuroprotective effects of oxygen‐glucose deprivation preconditioning (OGDPC). Immunocytochemistry was used to detect the expression of neurofilament in neurons. MTT and lactate dehydrogenase activity assay were used to measure the cytotoxicity. Western blot was used to detect the protein expression of BMP‐7 and downstream signals. BMP inhibitor, mitogen‐activated protein kinase inhibitors, Smad inhibitor and siRNA of Smad 1 were used to investigate the role of corresponding signaling pathways in the OGDPC. Results showed that OGDPC‐induced overexpression of BMP‐7 in primary cortical neurons and SH‐SY5Y cells. Both of endogenous and exogenous BMP‐7 could replicate the neuroprotective effects seen in OGDPC pretreatment. In addition, extracellular regulated protein kinases, p38 and Smad signaling pathway were found to be involved in the neuroprotective effects mediated by OGDPC via BMP‐7. This study primarily reveals the cellular mechanisms of the neuroprotection mediated by OGDPC, and provides evidence for better understanding of this intrinsic factor against ischemia. This article is protected by copyright. All rights reserved.
    September 19, 2015   doi: 10.1111/1440-1681.12492   open full text
  • Effect of common polymorphisms of the farnesoid X receptor and bile acid transporters on the pharmacokinetics of ursodeoxycholic acid.
    Miao Hu, Benny SP Fok, Siu‐Kwan Wo, Vincent HL Lee, Zhong Zuo, Brian Tomlinson.
    Clinical and Experimental Pharmacology and Physiology. September 18, 2015
    Ursodeoxycholic acid (UDCA), a natural, dihydroxy bile acid, promotes gallstone dissolution and has been attributed with several other beneficial effects. The farnesoid X receptor (FXR) may influence the pharmacokinetics of UDCA by modulating the expression of bile acid transporters. This exploratory study examined whether common functional polymorphisms in FXR and in bile acid transporter genes affect the pharmacokinetics of exogenous UDCA. Polymorphisms in genes for transporters involved in bile acid transport, solute carrier organic anion 1B1 (SLCO1B1) 388A>G and 521T>C, solute carrier 10A1 (SLC10A1) 800 C>T and ATP‐binding cassette B11 (ABCB11) 1331T>C, and the FXR ‐1G>T polymorphism were genotyped in 26 male Chinese subjects who ingested single oral 500‐mg doses of UDCA. Plasma concentrations of UDCA and its major conjugate metabolite glycoursodeoxycholic acid (GUDCA) were determined. The mean systemic exposure of UDCA was higher in the 5 subjects with one copy of the FXR ‐1G>T variant allele than in those homozygous for the wild‐type allele (n=21) (AUC0‐24h: 38.5±28.2 vs. 20.9±8.0 μg·h/ml, P = 0.021), but this difference appeared mainly due to one outlier with the ‐1GT genotype and elevated baseline and post‐treatment UDCA concentrations. After excluding the outlier, body weight was the only factor associated with plasma concentrations of UDCA and there were no significant associations with the other polymorphisms examined. None of the polymorphisms affected the pharmacokinetics of GUDCA. This study showed that the common polymorphisms in bile acid transporters had no significant effect on the pharmacokinetics of exogenous UDCA but an effect of the FXR polymorphism cannot be excluded. This article is protected by copyright. All rights reserved.
    September 18, 2015   doi: 10.1111/1440-1681.12490   open full text
  • CaMKIIδ and cardiomyocyte Ca2+ signaling – new perspectives on splice variant targeting.
    James R. Bell, Antonia J.A. Raaijmakers, Johannes V. Janssens, Lea M.D. Delbridge.
    Clinical and Experimental Pharmacology and Physiology. September 12, 2015
    Control of cardiomyocyte cytosolic Ca2+ levels is crucial in determining inotropic status and ischemia/reperfusion stress response. Responsive to fluctuations in cellular Ca2+, Ca2+/calmodulin‐dependent protein kinase II (CaMKII) is a serine/threonine kinase integral to the processes regulating cardiomyocyte Ca2+ channels/transporters. CaMKII is primarily expressed either in the δB or δC splice variant forms, which may mediate differential influences on cardiomyocyte function and pathological response mechanisms. Increases in myocyte Ca2+ levels promote the binding of a Ca2+/calmodulin complex to CaMKII, to activate the kinase. Activity is also maintained through a series of post‐translational modifications within a critical region of the regulatory domain of the protein. Our recent data indicate that the post‐translational modification status of CaMKIIδB/δC variants may have an important influence on reperfusion outcomes. We provided the first evidence that the specific type of CaMKII post‐translational modification has a role in determining target selectivity of downstream Ca2+ transporters. We were also able to demonstrate that the phosphorylated form of CaMKII closely co‐localises with CaMKIIδB in the nuclear/myofilament fraction, contrasting with a co‐enrichment of oxidized CaMKII in the membrane fraction with CaMKIIδC. We have also been able to conclude that a hyper‐phosphorylation of CaMKII (Thr287) in reperfused hearts represents a hyper‐activation of the CaMKIIδB, which exerts anti‐arrhythmic actions through an enhanced capacity to selectively increase sarcoplasmic reticulum Ca2+ uptake and maintain cytosolic Ca2+ levels. This suggests that suppression of global CaMKIIδ may not be an efficacious approach to developing optimal pharmacological interventions for the vulnerable heart. This article is protected by copyright. All rights reserved.
    September 12, 2015   doi: 10.1111/1440-1681.12489   open full text
  • Physicochemical Properties of Radiographic Contrast Media, Potential Nephrotoxicity and Prophylaxis.
    Barry Hogstrom, Nobuhiro Ikei.
    Clinical and Experimental Pharmacology and Physiology. September 10, 2015
    Contrast Induced Nephropathy remains a controversial topic. The clinical relevance of changes in laboratory parameters has been challenged; some authors have even suggested that CIN simply reflects natural fluctuations. Other areas of controversy include the pathophysiology of CIN, effectiveness of prophylactic approaches and differences in nephrotoxicity between individual CM. The aim of this review is to summarize our understanding of laboratory findings and explore their relationship to contrast media toxicity. This article is protected by copyright. All rights reserved.
    September 10, 2015   doi: 10.1111/1440-1681.12487   open full text
  • Icaritin inhibits the invasion and epithelial‐to‐mesenchymal transition of glioblastoma cells by targeting EMMPRIN via PTEN/AKt/HIF‐1α signaling.
    Bo Xu, Chuanwu Jiang, Hongxing Han, Hong Liu, Ming Tang, Longxi Liu, Wenyan Ji, Xuechao Lu, Xiuli Yang, Yunxu Zhang, Yongji Liu.
    Clinical and Experimental Pharmacology and Physiology. September 10, 2015
    Icaritin, a hydrolytic product of icariin from the Epimedium genus, exerts anti‐tumor effects on a variety of tumor cell types, mainly by inhibiting cell proliferation and inducing apoptosis. However, little is known about the role of icaritin in cancer invasion and epithelial‐to‐mesenchymal transition (EMT). In the present study, the glioblastoma (GBM) cell line U87MG was used as a model to investigate the effects of icaritin on the invasion and EMT of cancer cells. Our results showed that icaritin significantly inhibited the invasion and EMT of GBM cells by targeting extracellular matrix metalloproteinase (EMMPRIN). Furthermore, our findings strongly indicated that the modulatory effect of icaritin on EMMPRIN was mediated via the PTEN/Akt/HIF‐1α signaling pathway. Our data provide the first experimental evidence of the inhibitory effect of icaritin on cancer cell invasion and EMT, thus highlighting the potential of icaritin to be employed as a promising anti‐cancer agent in the treatment of GBM. This article is protected by copyright. All rights reserved.
    September 10, 2015   doi: 10.1111/1440-1681.12488   open full text
  • Effects of minocycline on parameters of cardiovascular recovery after cardioplegic arrest in a rabbit Langendorff heart model.
    Aida Salameh, Michelle Halling, Thomas Seidel, Stefan Dhein.
    Clinical and Experimental Pharmacology and Physiology. September 02, 2015
    Pharmacological cardiac organ protection during cardiopulmonary bypass presents an opportunity for improvement. A number of different strategies have been established to minimize ischemia/reperfusion‐induced damage to the heart. Among these, cardioplegia with histidine‐tryptophan‐ketoglutarate solution and hypothermia are the most frequently used regimen. The antibiotic minocycline has been used in this context for neuroprotection. The aim of our study was to evaluate whether the application of minocycline prior to cardioplegia exerts a protective effect on cardiac muscle. For this purpose, we investigated 6 rabbit hearts with minocycline treatment (1μmol/l) and 6 without in a Langendorff model of 90min cold cardioplegic arrest using Custodiol® followed by a 30min recovery phase. Histological analysis of cardiac muscle revealed that markers of apoptosis, oxidative and nitrosative stress were significantly lower in the minocycline group, whereas ATP‐ and MDA (malondialdehyde)‐levels and O2‐consumption were not affected by minocycline. Functionally, recovery of dP/dt(max) and dP/dt(min) was significantly faster in the minocycline group than in control. We conclude that adding minocycline to the cardioplegic solution may improve left ventricular recovery after cardioplegic arrest involving reduced pro‐apoptotic effects. This article is protected by copyright. All rights reserved.
    September 02, 2015   doi: 10.1111/1440-1681.12485   open full text
  • Poly(ADP‐ribose)Polymerase 1 Inhibition Protects Against Age‐Dependent Endothelial Dysfunction.
    Guang‐hao Zhang, Min Chao, Long‐hua Hui, Dong Ling Xu, We‐li Cai, Jie Zheng, Min Gao, Ming‐xiang Zhang, Juan Wang, Qing‐hua Lu.
    Clinical and Experimental Pharmacology and Physiology. August 31, 2015
    Age‐related endothelial dysfunction is closely associated with the local production of reactive oxygen species (ROS) within and in the vicinity of the vascular endothelium. Oxidant‐induced DNA damage can activate the nuclear enzyme poly(ADP‐ribose) polymerase 1 (PARP‐1), leading to endothelial dysfunction in various pathophysiological conditions. In the present study, we aimed to investigate the role of PARP‐1 in age‐dependent changes in endothelial cell function and its underlying mechanism. Wild‐type (WT) and PARP‐1‐/‐ mice were divided into young (2 months) and old (12 months) groups. Isolated aortic rings were suspended to record isometric tension to assess endothelial function. Nitric oxide (NO) production and content in plasma were detected by spectrophotometry. Superoxide (O2‐) production was detected by dihydroethidium. Expression of PARP‐1, endothelial nitric oxide synthase (eNOS), induced nitric oxide synthase (iNOS), and arginase‐2 (Arg2) was assessed by RT‐PCR and western blot analysis. Endothelium‐dependent relaxation in response to acetylcholine was lost in old WT, but not PARP‐1‐/‐, mice. Endothelium‐independent vasodilation was not impaired in aging mice. Production of O2‐ was greater in aging WT mice than young or aging PARP‐1‐/‐ mice. eNOS expression was not affected by aging in WT or PARP‐1‐/‐ mice, but p‐eNOS expression decreased and iNOS and Arg2 levels were upregulated only in aging WT mice. In conclusion, PARP‐1 inhibition may protect against age‐dependent endothelial dysfunction, potentially by regulating NO bioavailability via iNOS. Inhibition of PARP‐1 may help in vascular aging prevention. This article is protected by copyright. All rights reserved.
    August 31, 2015   doi: 10.1111/1440-1681.12484   open full text
  • Ectopic expression of Hoxb4a in hemangioblasts promotes hematopoietic development in early embryogenesis of zebrafish.
    Li‐Ping Shu, Zhi‐Wei Zhou, Ting Zhou, Min Deng, Mei Dong, Yi Chen, Yan‐Fang Fu, Yi Jin, Shu‐Feng Zhou, Zhi‐Xu He.
    Clinical and Experimental Pharmacology and Physiology. August 27, 2015
    Hemangioblast, including primitive hematopoietic progenitor cells, play an important role in hematopoietic development, however, the underlying mechanism for the propagation of hematopoietic progenitor cells remains elusive. A variety of regulatory molecules activated in early embryonic development play a critical role in the maintenance of function of hematopoietic progenitor cells. Homeobox transcription factors are an important class of early embryonic developmental regulators determining hematopoietic development. However, the effect of homeobox protein Hox‐B4 (HOXB4) ectopic expression on the development of hemangioblasts has not been fully addressed. This study aimed to investigate the role of Hoxb4a, an ortholog gene of HOXB4 in zebrafish, in hematopoietic development in zebrafish. A transgenic zebrafish line was established with Cre‐loxP system that stably overexpressed enhanced green fluorescent protein (EGFP)‐tagged Hoxb4a protein under the control of hemangioblast‐specific lmo2 promoter. Overexpression of Hoxb4a in the development of hemangioblasts resulted in a considerable increase in the number of stem cell leukemia (scl) and lmo2‐positive primitive hematopoietic progenitor cells occurring in the posterior intermediate cell mass (ICM). Interestingly, Hoxb4a overexpression also disrupted the development of myelomonocytes in the anterior yolk sac and the posterior ICM, without affecting erythropoiesis in the posterior ICM. Taken together, these results indicate that Hoxb4a favors the development of hematopoietic progenitor cells originated from hemangioblasts in vivo. This article is protected by copyright. All rights reserved.
    August 27, 2015   doi: 10.1111/1440-1681.12483   open full text
  • Autonomic modulation analysis in active and sedentary kidney transplanted recipients.
    Carlos José Moraes Dias, Luana Monteiro Anaisse Azoubel, Herikson Araújo Costa, Ednei Costa Maia, Bruno Rodrigues, Antonio Carlos Silva‐Filho, Maria Claúdia Irigoyen, Richard D Leite, Mário Sevilio de Oliveira, Cristiano Teixeira Mostarda, Carlos Alberto Alves Dias‐Filho.
    Clinical and Experimental Pharmacology and Physiology. August 18, 2015
    Modulation of the autonomic nervous system on heart rate can be compromised in chronic kidney disease and may result in changes in the frequency and duration of the cardiac cycle. The aim of this study was to evaluate autonomic modulation in active and sedentary renal transplant recipients. 20 renal‐transplanted individuals were analyzed at the Centro de Prevenção de Doenças Renais (Kidney Disease Education Center), in the academic hospital of Universidade Federal do Maranhão, divided into active group (AG) and sedentary group (SG). The AG group was in regular concurrent training intervention for eight weeks, involving six men and four women (age 43.10 ± 13.02) while the SG was composed of three men and seven women (age 36.8 ± 9.26). Analysis of heart rate variability in time and frequency domain demonstrated that HR mean values in the SG and AG groups were 787.32 ±79.60 and 870±106.66ms, respectively. Differences were observed in time domain (SDNN, RMSSD) and frequency domain (LF (%), HF (%) and LF/HF). Total index of LF (m²) and HF (m²) has shown no differences for the SG and AG groups. Biochemical variables presented significantly lower levels after eight weeks of training. Higher heart rate variability in time domain and greater vagal modulation was observed in the AG group. The AG group had greater vagal modulation when compared to SG, with removal of the sympathetic and increased parasympathetic in the behavior [A FRASE EM VERMELHO NÃO FEZ SENTIDO] was confirmed by sympatho‐vagal balance. The AG group also presented significant improvements in frequency domain. This article is protected by copyright. All rights reserved.
    August 18, 2015   doi: 10.1111/1440-1681.12481   open full text
  • Cyclo‐Gly‐Pro, a cyclic dipeptide, attenuates nociceptive behavior and inflammatory response in mice.
    Jamylle Nunes de Souza Ferro, Fernanda Lima Torres Aquino, Renan Guedes Brito, Priscila Laíse dos Santos, Jullyana de Souza Siqueira Quintans, Lucas Costa Souza, Almair Ferreira Araújo, Bruno Lourenço Diaz, Waldecy Lucca‐Júnior, Lucindo José Quintans‐Júnior, Emiliano Barreto.
    Clinical and Experimental Pharmacology and Physiology. August 16, 2015
    The present study aimed to investigate the antinociceptive and anti‐inflammatory effects of the cyclic dipeptide cyclo‐Gly‐Pro (CGP) in mice. Antinociceptive activity was assessed by employing different pain models, such as formalin test, acetic acid‐induced writhing, hot plate test, and carrageenan‐induced hyperalgesia, in mice. The number of c‐Fos‐immunoreactive cells in the periaqueductal gray (PAG) was evaluated in CGP‐treated mice. Anti‐inflammatory activity was evaluated using paw edema induced by carrageenan, compound 48/80, serotonin, and prostaglandin E2 (PGE2) and analyzed by plethysmometry. Quantitation of myeloperoxidase (MPO) in the paw was carried out to analyze the presence of neutrophils in tissue. Intraperitoneal injection of CGP produced a significant inhibition in both neurogenic and inflammatory phases of formalin‐induced pain. The antinociceptive effect of CGP, evaluated in the acetic acid‐induced writhing test, was detected for up to 6 h after treatment. Further, in the hot plate test, antinociceptive behavior was evoked by CGP, and this response was inhibited by naloxone. Animals treated with CGP did not present changes in motor performance. In CGP‐treated mice there was an increase in the number of c‐Fos‐positive neurons in the periaqueductal gray. In another set of experiments, CGP attenuated the hyperalgesic response induced by carrageenan. Furthermore, CGP also reduced the carrageenan‐increased MPO activity in paws. In addition, CGP also reduced the paw edema evoked by compound 48/80, serotonin, and PGE2. Taken together, these results may support a possible therapeutic application of the cyclic dipeptide cyclo‐Gly‐Pro toward alleviating nociception and damage caused by inflammation conditions. This article is protected by copyright. All rights reserved.
    August 16, 2015   doi: 10.1111/1440-1681.12480   open full text
  • Microvascular Resistance in Response to Iodinated Contrast Media in Normal and Functionally Impaired Kidneys.
    Osamu Kurihara, Masamichi Takano, Saori Uchiyama, Isamu Fukuizumi, Tetsuro Shimura, Masato Matsushita, Hidenori Komiyama, Toru Inami, Daisuke Murakami, Ryo Munakata, Takayoshi Ohba, Noritake Hata, Yoshihiko Seino, Wataru Shimizu.
    Clinical and Experimental Pharmacology and Physiology. August 16, 2015
    Contrast‐induced nephropathy (CIN) is considered to result from intrarenal vasoconstriction, and occurs more frequently in impaired than in normal kidneys. We hypothesised that iodinated contrast media would markedly change renal blood flow and vascular resistance in functionally impaired kidneys. We enrolled 36 patients (32 men; mean age, 75.3 ± 7.6 years) undergoing diagnostic coronary angiography and divided them into two groups based on the presence of chronic kidney disease (CKD), defined as an estimated glomerular filtration rate (eGFR) of < 60 mL/min/1.73 m2 (CKD and non‐CKD groups, n = 18 in both). Average peak velocity (APV) and renal artery resistance index (RI) were measured by Doppler flow wire before and after administration of the iodinated contrast media. APV and RI were positively and inversely correlated with the eGFR at baseline, respectively (APV: R = 0.545, P = 0.001; RI: R = ‐0.627, P < 0.001). Mean RI was significantly higher (P = 0.015) and APV was significantly lower (P = 0.026) in the CKD than in the non‐CKD group. APV (P < 0.001) and RI (P = 0.002) were significantly changed following contrast media administration in the non‐CKD group, but not in the CKD group (APV: P = 0.258; RI: P = 0.707). Although renal arterial resistance was higher in patients with CKD, but not affected by contrast media administration, suggesting that patients with CKD could have an attenuated response to contrast media. This article is protected by copyright. All rights reserved.
    August 16, 2015   doi: 10.1111/1440-1681.12479   open full text
  • 17‐(allylamino)‐17‐demethoxygeldanamycin drives Hsp70 expression, but fails to improve morphological or functional recovery in injured skeletal muscle.
    Cory W. Baumann, Jeffrey S. Otis.
    Clinical and Experimental Pharmacology and Physiology. August 16, 2015
    The stress inducible 70‐kDa heat shock protein (Hsp70) is instrumental to efficient morphological and functional recovery following skeletal muscle injury because of its roles in protein quality control and molecular signaling. Therefore, in attempt to improve recovery we increased Hsp70 expression with 17‐(allylamino)‐17‐demethoxygeldanamycin (17‐AAG) prior to and following an intramuscular injection of barium chloride (BaCl2) into the tibialis anterior (TA) of healthy young mice. To assess recovery, regenerating fiber cross‐sectional area (CSA) of the TA and in vivo peak isometric torque produced by the anterior crural muscles (TA, extensor digitorum longus and extensor hallucis muscles) were analyzed for up to 3 weeks after the injury. Because treatment of 17‐AAG and Hsp70 are known to influence inflammatory and myogenic signaling, we also assessed tumor necrosis factor‐α (TNF‐α) and myogenin content. We report that 17‐AAG was effective at up‐regulating Hsp70 expression, increasing content fivefold in the uninjured muscle. However, this significant increase in Hsp70 content did not enhance morphological or functional recovery following the injury, as the return of regenerating fiber CSA and in vivo peak isometric torque did not differ compared to that of the injured muscle from the vehicle treated mice. Treatment with 17‐AAG also altered TNF‐α and myogenin content, increasing both to a greater extent after the injury. Together, these findings demonstrate that although 17‐AAG may alter molecular makers of regeneration, it does not improve recovery following BaCl2‐induced skeletal muscle injury in healthy young mice. This article is protected by copyright. All rights reserved.
    August 16, 2015   doi: 10.1111/1440-1681.12477   open full text
  • Subcutaneous Pharmacokinetics of the Cardiac Hormone Vessel Dilator.
    Qingyu Zhou, Glenn Whelan, Shu‐Feng Zhou, Meghan L. Lane, David L. Vesely.
    Clinical and Experimental Pharmacology and Physiology. May 29, 2014
    Vessel dilator, a hormone synthesized in the heart, eliminates 71% of human small‐cell lung cancers and 67% of human breast cancers growing in mice when given subcutaneously via osmotic pumps. The pharmacokinetic evaluation of subcutaneously administered vessel dilator has never been performed. Pharmacokinetics of A) administering vessel dilator by a subcutaneous bolus(ScB), B) via a 3‐hour subcutaneous infusion (ScI) were compared with C) an intravenous(IV) bolus in male Fischer 344 rats. Half‐life (t½) of vessel dilator via ScI was 54 minutes while IV and ScB t1/2 were 43 and 30 minutes. tmax for vessel dilator via IV, ScB, and ScI were 1.5, 23 and 156 minutes while Cmax for vessel dilator was 3749, 887 and 471 ng/L (normalized) for IV, ScB and ScI. Area under the curve (AUC0‐∞) (ng h/mL) for vessel dilator was 1166, 880, and 1652 (normalized) for IV,ScB and ScI, respectively. The volume of distribution (L) for vessel dilator was 2.38, 0.92 and 1.08 for IV, ScB and SCI. Clearance was 1.69, 1.50 and 0.78 L/h for IV, ScB and SCI,. The plasma concentrations of vessel dilator after each of the 3 methods of treatment mirrored their predicted concentration‐time profiles. We conclude that vessel dillator via ScI but as a significantly greater AUC, t½ and slowed clearance compared to IV or ScB (P < 0.001), suggesting it is the preferred method based upon pharmacokinetics to treat cancers. This article is protected by copyright. All rights reserved.
    May 29, 2014   doi: 10.1111/1440-1681.12266   open full text
  • Additive effect of polymorphisms in the β2‐adrenoceptor and NADPH oxidase p22phox genes contributes to the loss of estimated glomerular filtration rate in Chinese.
    Tao Wang, Yan Zhang, JingTao Ma, Zhen Feng, Kai Niu, Bing Liu.
    Clinical and Experimental Pharmacology and Physiology. May 29, 2014
    Since increased oxidative stress may mediate the detrimental actions of the enhanced sympathetic nervous activity on the renal function and vice versa, we investigated the effect of the polymorphic Arg16Gly in the β2 adrenoceptor (ADRB2) gene, Trp64Arg in the ADRB3 gene and C242T in the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase p22phox gene on the estimated glomerular filtration rate (eGFR) in Chinese. Initially recruited from the different outpatient service of our hospitals, 668 individuals were finally eligible with complete demographic information. Laboratory tests were performed and eGFR was derived from the MDRD equation for the Chinese. Plasma norepinephrine level and genotype were determined by the high performance liquid chromatography and TaqMan method, respectively. Only across the Arg16Gly polymorphism did the eGFR show significant difference: it was lower in the Gly16Gly variation which also had the highest plasma norepinephrine level. This polymorphism still remained a significant determinant of the eGFR after the multivariate analysis. Of importance, the multifactor dimensionality reduction method further detected a significant synergism between the polymorphic Arg16Gly and C242T in the eGFR reduction. These observations clarified the effects of the studied polymorphisms on the eGFR and exemplified gene‐gene interaction influencing renal function. This article is protected by copyright. All rights reserved.
    May 29, 2014   doi: 10.1111/1440-1681.12268   open full text
  • Surgery‐induced changes in rat IL1ß and acetylcholine metabolism: role of physostigmine.
    Konstanze Plaschke, Ann‐Katrin Müller, Jürgen Kopitz.
    Clinical and Experimental Pharmacology and Physiology. May 29, 2014
    Pharmacological enhancement of cholinergic activities by administering physostigmine is known to induce protective effects. It is unclear, however, whether the impact of physostigmine on inflammation and acetylcholine metabolism is related to different types of surgical intervention or to anesthesia alone. To determine this, rats were subjected to partial liver resection (PLR) or sham surgery. A control group only received anesthesia. Half of each treatment group received a single intraoperative dose of physostigmine; the others received placebo. Acetylcholine esterase (ACHE) activity and IL1ß and acetylcholine (ACH) concentrations were determined. Both PLR and sham operation induced a time‐dependent increase in plasma concentration of IL1ß (3.9 and 4.8‐fold) as compared to rats that received anesthesia alone. In rat brain, IL1ß had increased by about twofold after surgery as compared to controls. Blood ACHE was transiently decreased after surgery. Brain ACHE activity increased 1.3‐fold (p=0.014) only after PLR; consequently, the cerebral ACH concentration was significantly reduced. Physostigmine administration significantly reduced IL1ß and ACHE levels. Cerebral ACH concentration was markedly increased from 543.9±121.5 (placebo) to 653.5±93.3 ng mg‐1 protein (p<0.001) after administering physostigmine. We conclude that a single dose of physostigmine intraoperatively had a sustained anti‐inflammatory effect up to 120 min after injection that was especially pronounced under the conditions of PLR surgery. In addition to its protective peripheral action, physostigmine exerts neuroprotective action by increasing levels of the neurotransmitter ACH. This article is protected by copyright. All rights reserved.
    May 29, 2014   doi: 10.1111/1440-1681.12267   open full text
  • Dual effects of amiodarone on pacemaker currents in hypertrophied ventricular myocytes isolated from spontaneously hypertensive rats.
    Hongxia Li, Yafeng Zhou, Bin Jiang, Xin Zhao, Xun Li, Xiangjun Yang, Wenping Jiang.
    Clinical and Experimental Pharmacology and Physiology. May 24, 2014
    The pacemaker current (If) conducted by hyperpolarization‐activated cyclic nucleotide‐gated (HCN) channels plays a critical role in the regulation of cardiac automaticity, with If density increased in hypertrophied ventricular myocytes. Amiodarone, a highly effective antiarrhythmic agent, blocks human HCN currents and native If under normal conditions. To determine the effects of amiodarone under pathologic conditions, we monitored If under both acute and chronic treatment conditions in ventricular myocytes from spontaneously hypertensive rats (SHR) with left ventricular hypertrophy using the whole‐cell patch‐clamp technique. If current density was significantly greater in SHR ventricular myocytes than in cells from healthy normotensive control Wistar‐Kyoto (WKY) rats. Acute application of amiodarone significantly decreased If density in both SHR and WKY myocytes. The inhibition was concentration‐dependent with an IC50 of 4.9 (±1.2) μM in SHR and 6.9 (±1.3) μM in WKY myocytes. Amiodarone increased the activation and deactivation times of If in SHR myocytes, although it did not alter the relationship of voltage‐dependent activation and the reversal potential of If in SHR myocytes. Chronic exposure of SHR myocytes to amiodarone potently inhibited If and downregulated HCN2 and HCN4, the major channel subtypes underlying native If, at both mRNA and protein levels. These findings indicate that amiodarone inhibits If under hypertrophied conditions through dual mechanisms: direct channel blockade of If currents and indirect suppression via negative regulation of HCN channel gene expression. These unique properties of amiodarone may contribute to its antiarrhythmic properties under pathologic conditions. This article is protected by copyright. All rights reserved.
    May 24, 2014   doi: 10.1111/1440-1681.12264   open full text
  • Enhanced external counterpulsation improves endothelial function and exercise capacity in patients With ischemic left ventricular dysfunction.
    DT Beck, JS Martin, DP Casey, JC Avery, PD Sardina, RW Braith.
    Clinical and Experimental Pharmacology and Physiology. May 24, 2014
    Enhanced external counterpulsation (EECP) therapy decreases angina episodes and improves quality of life in patients with left ventricular dysfunction (LVD). However, studies have not elucidated the mechanisms of action and overall effects of EECP in patients with LVD. The purpose of this study was to investigate the effects of EECP on endothelial function in peripheral conduit arteries and exercise capacity (peak VO2) in patients with LVD. Patients with ischemic LVD (EF 34.5±4.2%; n=9), and patients with symptomatic CAD and preserved LV function (EF 53.5±6.6%; n=15), were studied before and after 35 1‐hr sessions of EECP. Brachial (bFMD) and femoral (fFMD) artery flow‐mediated dilation were evaluated using high‐resolution ultrasound. EECP elicited similar significant improvements in the following FMD parameters amongst the CAD and LVD groups, respectively: absolute bFMD (+53% and +70%); relative bFMD (+50% and +74%); bFMD normalized for shear rate (+70% and +61%); absolute fFMD (+33% and +21%); and relative fFMD (+32% and +17%) (P≥0.05 between groups). EECP significantly improved plasma levels of nitrate/nitrite (NOx) (+55% and +28%; μmol/L) and prostacyclin (6‐keto‐PGF1α) (+50% and +70%); and improved peak VO2 (+36% and +21%), similarly in both the CAD and LVD groups, respectively; (P≥0.05 between groups). Despite reduced LV function, EECP therapy significantly improves peripheral vascular function and functional capacity similar in magnitude to that observed in CAD patients with preserved LV function. This article is protected by copyright. All rights reserved.
    May 24, 2014   doi: 10.1111/1440-1681.12263   open full text
  • Metformin may produce antidepressant effects through improvement of cognitive function among depressed patients with diabetes mellitus.
    Min Guo, Jia Mi, Qiu‐Ming Jiang, Jin‐Mei Xu, Ying‐Ying Tang, Geng Tian, Bin Wang.
    Clinical and Experimental Pharmacology and Physiology. May 24, 2014
    Diabetes mellitus and depressive disorders are both common chronic diseases that increase functional disability and social burden. Cognitive impairment is a potentially debilitating feature of depression. Previous evidence has revealed that the antidiabetic drug metformin could be suitable for diabetic patients with cognitive impairment. However, there is no direct evidence from clinical studies that metformin treatment improves cognitive function in diabetic patients suffering from depression. In this study 58 participants, diagnosed with depression and type 2 diabetes mellitus, were recruited and divided into two groups: one treated with metformin and one with placebo for 24 weeks. Cognitive function, depressive behavior and diabetes improvement were evaluated. Chronic treatment with metformin for 24 weeks improved cognitive performance assessed by the Wechsler Memory Scale‐Revised (WMS‐R) in depressed patients with type 2 diabetes mellitus. In addition, metformin significantly improved depressive performance and changed the glucose metabolism in depressed patients with diabetes. Depressive symptoms negatively correlated with the cognitive performance in metformin‐treated participants. Furthermore, associations were also observed between the parameters of blood glucose metabolism and the depression phenotype. These findings suggested that chronic treatment with metformin produced antidepressant behavioral effects, and that improved cognitive function is involved in the therapeutic outcome of metformin. The current results also raised the possibility that supplementary administration of antidiabetic medications might enhance the recovery of depression, co‐morbid with type 2 diabetes mellitus, through improvements in cognitive performance. This article is protected by copyright. All rights reserved.
    May 24, 2014   doi: 10.1111/1440-1681.12265   open full text
  • Monte Carlo simulation analysis of ceftobiprole, dalbavancin, daptomycin, tigecycline, linezolid and vancomycin pharmacodynamics against intensive care unit‐isolated methicillin‐resistant Staphylococcus aureus.
    Ahmed Hamed Salem, George G Zhanel, Safaa A Ibrahim, Ayman M Noreddin.
    Clinical and Experimental Pharmacology and Physiology. May 20, 2014
    The aim of the present study was to compare the potential of ceftobiprole, dalbavancin, daptomycin, tigecycline, linezolid and vancomycin to achieve their requisite pharmacokinetic/pharmacodynamic (PK/PD) targets against methicillin‐resistant Staphylococcus aureus isolates collected from intensive care unit (ICU) settings. Monte Carlo simulations were carried out to simulate the PK/PD indices of the investigated antimicrobials. The probability of target attainment (PTA) was estimated at minimum inhibitory concentration values ranging from 0.03 to 32 μg/mL to define the PK/PD susceptibility breakpoints. The cumulative fraction of response (CFR) was computed using minimum inhibitory concentration data from the Canadian National Intensive Care Unit study. Analysis of the simulation results suggested the breakpoints of 4 μg/mL for ceftobiprole (500 mg/2 h t.i.d.), 0.25 μg/mL for dalbavancin (1000 mg), 0.12 μg/mL for daptomycin (4 mg/kg q.d. and 6 mg/kg q.d.) and tigecycline (50 mg b.i.d.), and 2 μg/mL for linezolid (600 mg b.i.d.) and vancomycin (1 g b.i.d. and 1.5 g b.i.d.). The estimated CFR were 100, 100, 70.6, 88.8, 96.5, 82.4, 89.4, and 98.3% for ceftobiprole, dalbavancin, daptomycin (4 mg/kg/day), daptomycin (6 mg/kg/day), linezolid, tigecycline, vancomycin (1 g b.i.d.) and vancomycin (1.5 g b.i.d.), respectively. In conclusion, ceftobiprole and dalbavancin have the highest probability of achieving their requisite PK/PD targets against methicillin‐resistant Staphylococcus aureus isolated from ICU settings. The susceptibility predictions suggested a reduction of the vancomycin breakpoint to 1 μg/mL.
    May 20, 2014   doi: 10.1111/1440-1681.12195   open full text
  • Intradermal glutamate and capsaicin injections: Intra‐ and interindividual variability of provoked hyperalgesia and allodynia.
    Matias Nilsson, Dorte Lassen, Trine Andresen, Anders K Nielsen, Lars Arendt‐Nielsen, Asbjørn M Drewes.
    Clinical and Experimental Pharmacology and Physiology. May 20, 2014
    Intradermal injections of glutamate and capsaicin are attractive to use in human experimental pain models because hyperalgesia and allodynia mimic isolated aspects of clinical pain disorders. The aim of the present study was to investigate the reproducibility of these models. Twenty healthy male volunteers (mean age 24 years; range 18–38 years) received intradermal injections of glutamate and capsaicin in the volar forearm. Magnitudes of secondary pinprick hyperalgesia and brush‐evoked allodynia were investigated using von Frey filaments (gauges 10, 15, 60 and 100 g) and brush strokes. Areas of secondary hyperalgesia and allodynia were quantified immediately after injection and after 15, 30 and 60 min. Two identical experiments separated by at least 7 days were performed. Reproducibility across and within volunteers (inter‐ and intra‐individual variation, respectively) was assessed using intraclass correlation coefficient (ICC) and coefficient of variation (CV). Secondary pinprick hyperalgesia was observed as a marked increase in the visual analogue scale (VAS) response to von Frey gauges 60 and 100 g (P < 0.001) after glutamate injection. For capsaicin, secondary pinprick hyperalgesia was detected with all von Frey gauges (P < 0.001). Glutamate evoked reproducible VAS response to all von Frey gauges (ICC > 0.60) and brush strokes (ICC > 0.83). Capsaicin injection was reproducible for secondary hyperalgesia (ICC > 0.70) and allodynia (ICC > 0.71). Intra‐individual variability was generally lower for the VAS response to von Frey and brush compared with areas of secondary hyperalgesia and allodynia. In conclusion, glutamate and capsaicin yield reproducible hyperalgesic and allodynic responses, and the present model is well suited for basic research, as well as for assessing the modulation of central phenomena.
    May 20, 2014   doi: 10.1111/1440-1681.12229   open full text
  • Naringenin attenuates CCl4‐induced hepatic inflammation by the activation of an Nrf2‐mediated pathway in rats.
    Mohammad Ali Esmaeili, Mostafa Alilou.
    Clinical and Experimental Pharmacology and Physiology. May 20, 2014
    The possible protective effects of naringenin, a naturally occurring citrus flavonone, on carbon tetrachloride (CCl4)‐induced liver injury in rats and the mechanism underlying its effects were investigated. Forty rats were divided into five groups. Rats in Groups I and II served as the normal and injured liver groups, respectively; Group III rats were treated with the standard drug silymarin as a positive control; and rats in Groups IV and V (naringenin‐treated groups) were administrated 50 mg/kg, p.o., naringenin for 7 days. Liver samples were collected to evaluate mRNA and protein expression, histological changes and oxidative stress. Naringenin inhibited lipid peroxidation and reduced serum levels of hepatic enzymes induced by CCl4. In addition, naringenin increased the liver content of reduced glutathione and the activity of anti‐oxidant enzymes in rats treated with CCl4. Naringenin attenuated liver inflammation by downregulating CCl4‐induced activation of tumour necrosis factor (TNF)‐α, inducible nitric oxide synthase (iNOS) and cyclo‐oxygenase (COX‐2) at both the protein and mRNA levels. Naringenin treatment significantly increased NF‐E2‐related factor 2 (Nrf2) and heme oxygenase (HO‐1) expression in injured livers. In rats treated with CCl4 alone, decreases were seen in nuclear Nrf2 expression and in the mRNA levels of its target genes (e.g. HO‐1, NQO1 and glutathione S‐transferase alpha 3 (GST‐a3)). Together, the results suggest that naringenin can protect the liver against oxidative stress, presumably by activating the nuclear translocation of Nrf2 as well as attenuating the TNF‐α pathway to elicit an anti‐inflammatory response in liver tissue.
    May 20, 2014   doi: 10.1111/1440-1681.12230   open full text
  • Association of HMGB1 and HMGB2 genetic polymorphisms with lung cancer chemotherapy response.
    Ying Wang, Xiang‐Ping Li, Ji‐Ye Yin, Yu Zhang, Hui He, Chen‐Yue Qian, Juan Chen, Yi Zheng, Kamila Smieszkol, Yi‐Lan Fu, Zi‐Yu Chen, Hong‐Hao Zhou, Zhao‐Qian Liu.
    Clinical and Experimental Pharmacology and Physiology. May 20, 2014
    The aim of the present study was to investigate the association of genetic polymorphisms in high mobility group box 1 and 2 (HMGB1 and HMGB2, respectively) with platinum‐based chemotherapy responses in Chinese lung cancer patients. In total, 338 Chinese lung cancer patients (154 responders and 184 non‐responders) were recruited to the study. All patients received at least two cycles of first‐line platinum‐based chemotherapy. Three tagging single nucleotide polymorphisms (SNPs) of HMGB1 and two tagging SNPs of HMGB2 were detected in patients. We found that rs1412125 and rs2249825 of HMGB1 were significantly associated with the platinum‐based chemotherapy response in both recessive and genotypic models. In addition, rs1412125 showed significant association with platinum‐based chemotherapy response for the subgroup of patients aged >55 years in additive, recessive and genotypic models. No significant associations were detected between other SNPs and the platinum‐based chemotherapy response. The HMGB1 SNPs (rs1412125 and rs2249825) were associated with platinum‐based chemotherapy responses in Chinese lung cancer patients. In conclusion, HMGB1 SNPs may serve as potential biomarkers for predicting the efficacy of platinum‐based chemotherapy.
    May 20, 2014   doi: 10.1111/1440-1681.12232   open full text
  • Oestrogen upregulates the sarcoplasmic reticulum Ca2+ ATPase pump in coronary arteries.
    Brent JF Hill, Edwin Muldrew.
    Clinical and Experimental Pharmacology and Physiology. May 20, 2014
    The presence of circulating plasma 17β‐oestradiol (E2) is beneficial in women against abnormal vascular tone development, such as coronary arterial vasospasms. Several vascular diseases have demonstrated that increased expression of the sarcoplasmic reticulum Ca2+‐ATPase pump (SERCA2b) serves to limit the excessive accumulation of intracellular Ca2+. Therefore, the hypothesis of the present study was that E2 would increase SERCA2b expression in the coronary vasculature. Coronary arteries were dissected from hearts obtained from mature female pigs. Artery segments were cultured for 24 h in E2 (1 pmol/L or 1 nmol/L) and homogenized for western blot analysis. At 1 nmol/L, E2 induced an approximate 50% increase in immunoreactivity for SERCA2b. In addition, E2 increased the protein expression of the known SERCA regulatory proteins, protein kinase A (PKA) and protein kinase G (PKG). The E2‐induced increase in SERCA2b was attenuated when the culture medium was supplemented with the oestrogen receptor (ER) α/β antagonist ICI 182,780 and the PKG antagonist KT5823 (10 μmol/L, 24 h for both). The PKA antagonist (KT5720; 10 μmol/L, 24 h) had no effect on SERCA2b expression. Removal of the endothelium (using a wooden toothpick) from artery segments prior to culture decreased the E2‐mediated increase in SERCA2b and PKG expression by 45% and 47%, respectively. Overall, the findings suggest that one of the potential cardiovascular benefits of E2 in women is upregulation of SERCA2b, via activation of the classic ERα and ERβ pathway.
    May 20, 2014   doi: 10.1111/1440-1681.12233   open full text
  • Age‐ and sex‐related differences in sudomotor function evaluated by the quantitative sudomotor axon reflex test (QSART) in healthy humans.
    Jeong Beom Lee, In Ho Lee, Young Oh Shin, Young Ki Min, Hun Mo Yang.
    Clinical and Experimental Pharmacology and Physiology. May 20, 2014
    The aim of the present study was to quantitatively investigate the age and sex‐related differences in sudomotor function in healthy humans. The quantitative sudomotor axon reflex test (QSART) with iontophoresis (2 mA for 5 min) and 10% acetylcholine (ACh) was performed to determine axon reflex‐mediated (AXR), with and without iotophoresis (AXR(1) and AXR(2), respectively), and directly activated (DIR) sweating. All experiments were conducted under thermoneutral conditions (temperature 24.0 ± 0.5°C; relative humidity 40 ± 3%). In general, men had enhanced values of onset time of AXR, sweat rates, activated sweat gland density (SGD) and activated sweat gland output (SGO) than women, but not in all cases. The onset time of AXR (r2 = 0.567; P < 0.001) was positively correlated with advancing age, whereas sweat rates of AXR(1) and AXR(2) (r2 = 0.571 and r2 = 0.486, respectively; P < 0.0001), DIR (r2 = 0.594; P < 0.0001), SGD (r2 = 0.496; P < 0.0001) and SGO (r2 = 0.551; P < 0.0001) were negatively correlated in both men and women with advancing age. The results demonstrate that an attenuation of sudomotor function occurs with aging in both sexes. Moreover, the findings showed a progressive increase in onset time and a decrease in sweat rates, SGD and SGO with increasing age in both sexes. A variation in sweat function was found between sexes, but not in all age groups.
    May 20, 2014   doi: 10.1111/1440-1681.12234   open full text
  • Association of nitric oxide synthase 3 (NOS3) 894 G>T polymorphism with prognostic outcomes of anthracycline in Chinese patients with acute myeloid leukaemia.
    Hui He, Ya‐Jing Xu, Ji‐Ye Yin, Xi Li, Jian Qu, Xiao‐Jing Xu, Zhuo‐Gang Liu, Fan Zhou, Ming Zhai, Yan Li, Hong‐Hao Zhou, Zhao‐Qian Liu.
    Clinical and Experimental Pharmacology and Physiology. May 20, 2014
    The aim of the present study was to investigate the influence of the nitric oxide synthase 3 (NOS3) 894 G>T polymorphism on prognostic outcomes of anthracycline in Chinese patients with de novo intermediate‐risk acute myeloid leukaemia (AML) and to examine the gene expression level in relation to genetic variation. In all, 225 Chinese patients with intermediate‐risk AML (at the complete remission stage) treated with anthracycline were enrolled in the study. The 894 G>T polymorphism of the NOS3 gene was analysed by allele‐specific matrix‐assisted laser desorption ionization time‐of‐flight. Expression of NOS3 mRNA was tested in 72 patients of known genotype for NOS3 894 G>T. The clinical characteristics of these patients were obtained from medical records. Survival analysis showed that patients with AML (GG genotype) had a longer overall survival (OS; P = 0.006). After adjusting for age, gender, leucocyte count, haemoglobin level, platelet level, French, American and Britain (FAB) classification, lactate dehydrogenase levels, Eastern Cooperative Oncology Group Performance Status, nucleophosmin gene and fms‐related tyrosine kinase 3 gene, multivariate survival analysis showed that the NOS3 894 G>T polymorphism appeared to be a predicting factor for OS (P = 0.014; hazard ratio = 1.856). However, no significant associations between the NOS3 894 G>T polymorphism and relapse‐free survival and relapse in patients with AML were observed. Gene expression levels were significantly higher in patients with the GG genotype than in patients with the GT and TT genotypes (P = 0.033). The findings suggest that the NOS3 894 G>T variant may be a biomarker for the prediction of OS in Chinese patients with AML.
    May 20, 2014   doi: 10.1111/1440-1681.12235   open full text
  • The Florey turns 50.
    John P Coghlan.
    Clinical and Experimental Pharmacology and Physiology. May 20, 2014
    The origins of the Howard Florey Laboratories of Experimental Physiology, Department of Physiology, The University of Melbourne, are tied to the ground‐breaking clinical work of Derek A Denton in 1947 and to the investigations of the initial scientific team into the control of salt and water balance in health and disease over the period 1947–1963 were Professor RD Wright, Drs JR Goding, IR McDonald, John P Coghlan, E Marelyn Wintour and John R Blair‐West. An Act of Parliament in 1971 by the Victorian State Government formally established the Institute named after Howard Florey, the Australian Nobel Prize winner who isolated penicillin. The Howard Florey Laboratories/Institute quickly became an international leader in the scientific areas of the physiological control of body fluids and electrolyte balance, especially sodium regulation and the regulation of the secretion of aldosterone, the adrenal salt‐retaining hormone; the micro measurement of hormones, in particular steroids and peptides; instinctive ingestive behaviour; fetal fluid regulation; hybridization histochemistry, and the hormone relaxin. Subsequently, the senior staff included, inter alia, Bruce Scoggins, Richard Weisinger, John McDougall, Brian Oldfield, Michael McKinley, Robin McAllen, Hugh Niall, Geoff Tregear and Felix Beck. During the 1990s, an explosion occurred in neuroscience and, in 1997, the Board made the strategic decision to change the focus of the Institute to brain disorders. From 1997 to 2007, Fred Mendelsohn steered the Florey to become one of Australia's premier brain research institutes and, under the current director (the eminent clinician and neuroscientist Geoffrey Donnan), this reputation has been further enhanced.
    May 20, 2014   doi: 10.1111/1440-1681.12236   open full text
  • Increased carotid plaque burden in men with the fibrillin‐1 2/3 genotype.
    Rachel Basso, Bo Hedblad, Joyce Carlson, Margaretha Persson, Gerd Östling, Toste Länne.
    Clinical and Experimental Pharmacology and Physiology. May 17, 2014
    Fibrillin‐1 is an important constituent of the vascular wall and earlier studies have indicated an effect of the Fibrillin‐1 (FBN1) 2/3 genotype on blood pressure as well as aortic stiffness in men. The aim was to determine if the FBN1 2/3 genotype was associated with presence of carotid plaque and incident cardiovascular morbidity and mortality in middle‐aged subjects. The FBN1 genotype was characterized in 5765 subjects (2424 men, 3341 women; aged 45‐69 years) recruited from the Malmö Diet and Cancer Study Cardiovascular Cohort, Sweden. Plaque occurrence and intima media thickness (IMT) of the carotid artery were assessed by ultrasound. Incidence of first cardiovascular events (myocardial infarction and stroke) and cause‐specific mortality was monitored during a mean of 13.2 years follow‐up. The most common FBN1 genotypes were 2/2, 2/3 and 2/4 which accounted for 92.2% (n=5317) of the subjects. There were no differences between the three genotypes regarding age, blood pressure, glucose, lipids, smoking habits, CCA diameter and IMT in men and women. Presence of plaque in the carotid artery was higher in men with genotype 2/3 as compared to the 2/2 and 2/4 genotypes, (55% vs. 46% and 50%, P=0.007). No similar difference was observed in women. No significant relationship was observed between FBN1 genotypes and incidence of cardio vascular disease or all‐cause mortality. The increased prevalence of plaque in the carotid artery of middle‐aged men with FBN1 2/3 genotype indicates a pathological arterial wall remodelling with a more pronounced atherosclerotic burden. This article is protected by copyright. All rights reserved.
    May 17, 2014   doi: 10.1111/1440-1681.12259   open full text
  • Nonalcoholic fatty liver disease (NAFLD) fibrosis score predicts 6.6 years overall mortality of Chinese patients with NAFLD.
    Yun‐hao Xun, Jian‐chun Guo, Guo‐qiang Lou, Yan‐ming Jiang, Zhen‐jie Zhuang, Meng‐fei Zhu, Yan Luo, Xiao‐jie Ma, Jing Liu, Dong‐xue Bian, Jun‐ping Shi.
    Clinical and Experimental Pharmacology and Physiology. May 17, 2014
    Nonalcoholic fatty liver disease (NAFLD) fibrosis score (NFS) has emerged as a useful predictor for a long‐term outcome in NAFLD patients. We evaluated the predictive performance of NFS for overall mortality in a Chinese population with NAFLD. All ultrasonography‐diagnosed NAFLD patients at our center between 1996 and 2011 were retrospectively recruited. The outcome was obtained by interview and the causes of death were confirmed by medical records. Area under receiver operating characteristic curve (AUC) was used to determine the predictive accuracy of NFS, BARD, FIB‐4 and aspartate aminotransferase/platelet ratio index (APRI) for mortality. Data from total of 180 eligible patients (median age 39 years; 96 males) were analyzed, with 12 deaths over a median follow‐up of 6.6 (range: 0.5‐14.8) ‐year. By Cox model analysis, NFS as a continuous variable other than 3 other scoring systems was identified as the only predictor for all‐cause mortality [hazard ratio 2.743, 95% confidential interval (CI) 1.670~4.504]. NFS yielded the highest AUC of 0.828 (95% CI 0.728~0.928, p<0.05), followed by FIB‐4, APRI and BARD with their AUCs of 0.806, 0.732 and 0.632 respectively (p<0.05, except for BARD). Collectively, NFS is a useful predictor of 6.6‐year all‐cause mortality for Chinese patients with NAFLD. This article is protected by copyright. All rights reserved.
    May 17, 2014   doi: 10.1111/1440-1681.12260   open full text
  • Effect of Long‐Term Application of Metoprolol and Propranolol in a Rat Model of Smoking.
    Yujiao Zhou, Ming Xu, Yuan Zhang, Yang Guo, Youyi Zhang, Bei He.
    Clinical and Experimental Pharmacology and Physiology. May 17, 2014
    β‐blockers, especially selective β1‐blockers, are often used to treat cardiovascular disease, even when complicated by chronic obstructive pulmonary disease. The association of β‐blocker selectivity and treatment effects is disputed, and the curative as well as side effects of various antagonists may differ. We investigated the effect of 1‐month treatment with the selective β1‐blocker metoprolol and nonselective β‐blocker propranolol on pulmonary function and pathology in a 4‐month rat model of passive cigarette‐smoke exposure and explored a potential mechanism. Lung function and general pathological changes were evaluated after 4‐month cigarette‐smoke exposure, with metoprolol and propranolol treatment during the last month. The levels of cytokines and mucin in bronchoalveolar lavage fluid were detected by ELISA. β1 and β2‐adrenergic receptor expression in the lung was detected by immunohistochemistry and western blot analysis. Chronic treatment with metoprolol and propranolol did not exacerbate peak expiratory flow or intra‐airway pressure in rats exposed to cigarette smoke. Propranolol significantly attenuated inflammatory cell infiltration, cytokine levels (tumor necrosis factor α and interleukin 8) in bronchoalveolar lavage fluid or mucus secretion; metoprolol reduced only smooth muscle proliferation. Moreover, propranolol treatment was associated but not significantly with restoring β2‐adrenergic receptor expression in the airway epithelia. Propranolol had a more beneficial effect on cigarette smoking‐induced lung damage than metoprolol in a smoking rat model and may be involved in restoring endogenous β2‐adrenergic receptor density in the airway epithelial cells. This article is protected by copyright. All rights reserved.
    May 17, 2014   doi: 10.1111/1440-1681.12261   open full text
  • Emodin enhances cholesterol efflux through activation of peroxisome proliferator‐activated receptor‐gamma in oxidized low density lipoprotein‐loaded THP‐1 macrophages.
    Xin Fu, Ai‐guo Xu, Meng‐ying Yao, Li Guo, Luo‐sha Zhao.
    Clinical and Experimental Pharmacology and Physiology. May 17, 2014
    Peroxisome proliferator‐activated receptors‐gamma (PPAR‐γ) is a nuclear receptor involved in the regulation of lipid metabolism. In the present study, we sought to investigate the effects of emodin, an anthraquinone derivative isolated from the roots of Rheum palmatum, on PPAR‐γ signaling and cholesterol efflux in macrophage foam cells. Oxidized low density lipoprotein (oxLDL)‐stimulated THP‐1 macrophages were incubated with different concentrations of emodin (0‐10 μM) for 18 h. Western blot analysis and semi‐reverse transcription polymerase chain reaction were used to assess the expression of key genes involved in cholesterol efflux including PPAR‐γ, liver X receptor alpha (LXR‐α), ATP‐binding cassette transporter A1 (ABCA1), and ABCG1. The levels of apolipoprotein A‐I (apoA‐I)‐mediated cholesterol efflux in emodin‐treated cells were measured. PPAR‐γ mRNA and protein levels were increased in emodin‐treated cells in a time and dose‐dependent manner. Emodin treatment also resulted in a concentration‐dependent induction of LXR‐α, ABCA1, and ABCG1 expression. Moreover, emodin promoted apoA‐I‐mediated cholesterol efflux from oxLDL‐loaded THP‐1 macrophages, which were significantly abolished by pretreatment with the PPAR‐γ selective antagonist GW9662 or specific small interfering RNA for PPAR‐γ. Taken together, our results demonstrate that emodin promotes cholesterol efflux from THP‐1 macrophages through activation of the PPAR‐γ signaling pathway and may represent a potential anti‐atherosclerotic drug. This article is protected by copyright. All rights reserved.
    May 17, 2014   doi: 10.1111/1440-1681.12262   open full text
  • Estrogen compromises the facilitatory effect of chronic nicotine on adenosine A2B receptor/K+ channel‐mediated renal vasodilations.
    Mahmoud M. El‐Mas, Sahar M. El‐Gowilly, Lamia K. Elsalakawy, Hanan M. El‐Gowelli.
    Clinical and Experimental Pharmacology and Physiology. May 14, 2014
    We have previously shown that the renal vasodilatory action of the adenosine analog 5′‐N‐ethyl carboxamidoadenosine (NECA) in female rats is mediated via preferential activation of adenosine A2B receptors (A2BRs)/K+ channel signaling. In this study, we tested the hypothesis that the renal vasodilatory effect of NECA and its A2BRs/K+ channel specificities are altered by chronic nicotine administration. The estrogenic modulation of the nicotine‐NECA renovascular interaction was also evaluated by determining the effect of ovariectomy (OVX) and estrogen replacement (OVXE2) on the evoked responses. In the isolated phenylephrine‐preconstricted perfused kidneys obtained from sham‐operated rats, vasodilations caused by cumulative bolus injections of NECA (1.6–50 nmol) or papaverine (1‐243 nmol) were not affected by nicotine (1‐8 mg/kg/day i.p., 2 weeks). NECA, but not papaverine, vasodilations were reduced in kidneys of OVX rats and restored to near‐sham values after E2 replacement. Further, nicotine increased NECA vasodilations in perfused kidneys obtained from OVX rats while it failed to do so in OVXE2 preparations. The enhanced NECA responsiveness in nicotine‐treated OVX preparations was abolished after the infusion of alloxazine (A2BR antagonist) or BaCl2 plus glibenclamide (blockers of inward rectifier and ATP‐sensitive K+ channels, respectively). Vasodilations caused by minoxidil (K+ channel opener) were increased by nicotine in OVX, but not OVXE2, preparations and this increase was abolished after BaCl2/glibenclamide infusion. Together, the data suggest that chronic nicotine enhances the A2BR/K+ channel‐mediated renal vasodilations in estrogen depleted rats. This article is protected by copyright. All rights reserved.
    May 14, 2014   doi: 10.1111/1440-1681.12255   open full text
  • Impacts of eNOS rs1799983 and ACE rs4646994 polymorphisms on the therapeutic efficacy of salvianolate injection in Chinese patients with coronary heart disease.
    Chao Fang, Xian Ren, Huan Zhou, Zhi‐Cheng Gong, Li Shen, Jing Bai, Ji‐Ye Yin, Jian Qu, Xiang‐Ping Li, Hong‐Hao Zhou, Zhao‐Qian Liu.
    Clinical and Experimental Pharmacology and Physiology. May 14, 2014
    The purpose of this study was to clarify the association of eNOS 894 G/T and ACE I/D genetic polymorphisms with risk of coronary heart disease (CHD) and to explore the effects of these polymorphisms on the therapeutic efficacy of salvianolate injection in Chinese CHD patients. 153 CHD patients and 198 healthy controls were enrolled in this study. We collected 5ml peripheral blood to extract DNA. Genetic diagnosis of eNOS 894G/T was determined by direct sequencing. Polymerase chain reaction‐restriction fragment length polymorphism analysis (PCR‐RFLP) was used to detect ACE I/D genotypes. We observed significant differences in the frequency distribution of eNOS and ACE polymorphisms between CHD patients and healthy controls (p<0.05). Binary logistic regression stepwise analysis revealed that the genotypes had an additive and dominant effect on patients’ therapeutic response (p<0.05). These data suggested that the genetic polymorphisms of ACE I/D and eNOS 894G/T probably play a role in the development of CHD and these genetic polymorphisms might affect the response to salvianolate injection in Chinese CHD patients. This article is protected by copyright. All rights reserved.
    May 14, 2014   doi: 10.1111/1440-1681.12257   open full text
  • The Novel Role and Underlying Mechanism of Wnt5a in Regulating Cellular Cholesterol Accumulation.
    Li Qin, Rong Hu, Neng Zhu, Hai‐Lun Yao, Xiao‐Yong Lei, Shun‐Xiang Li, Duan‐Fang Liao, Xi‐Long Zheng.
    Clinical and Experimental Pharmacology and Physiology. May 14, 2014
    Cholesterol accumulation is a critical step during the development and progression of atherosclerosis (As). Recently, the expression of Wnt5a has been found to be strikingly upregulated in both murine and human atherosclerotic lesions. However, the effect and mechanism of Wnt5a in atherosclerosis is poorly understood. In the present study, we investigated the effects and potential mechanisms of Wnt5a on cholesterol accumulation during atherosclerosis. We utilized RAW264.7 and vascular smooth muscle cells (VSMCs) treated with ox‐LDL as lipid‐loaded cell models. We found that the expression of Wnt5a protein was increased in a concentration‐dependent and time‐dependent manner under ox‐LDL treatment. To explore the underlying mechanism, we used Wnt5a siRNA to knockdown its expression or applied recombinant Wnt5a (rWnt5a) to stimulate its signaling. After knocking down Wnt5a, the content of total cholesterol (TC) and free cholesterol (FC) increased significantly (P<0.05) upon exposure of the cells to ox‐LDL. Conversely, the content of TC and FC decreased dramatically (P<0.05) after treatment with rWnt5a. More importantly, the expression of Caveolin‐1 and ABCA1 was dramatically reduced upon exposure to ox‐LDL when Wnt5a was knocked down; whereas, in the cells treated with rWnt5a, the expression of these proteins was increased significantly upon the exposure to ox‐LDL. Taken together, these findings demonstrate for the first time that Wnt5a reduces the accumulation of cholesterol in lipid‐loaded cells through regulation of the expression of Caveolin‐1 and ABCA1 at the mRNA levels, which are involved in reverse cholesterol transport. This may present a novel mechanism of Wnt5a‐mediated cholesterol transportation in macrophages and VSMCs. Therefore, targeting the Wnt5a signaling pathway may have clinical implications in atherosclerosis. This article is protected by copyright. All rights reserved.
    May 14, 2014   doi: 10.1111/1440-1681.12258   open full text
  • Characterization of thrombogenic, endothelial and inflammatory markers in supraventricular tachycardia: a study in patients with structurally normal hearts.
    Carlee D Schultz, Geetanjali Rangneker, Han S Lim, Angelique Fraudeau, Glenn Young, Kurt Roberts‐Thomson, Bobby John, Matthew Worthley, Prashanthan Sanders, Scott R Willoughby.
    Clinical and Experimental Pharmacology and Physiology. May 14, 2014
    Patients with atrial fibrillation (AF) are at an increased risk of thromboembolism and stroke primarily from the development of thrombi within the left atrial. Pathological changes in blood constituents and atrial endothelial damage promote the left atrial thrombus formation. It is not known whether factors predisposing to left atrial thrombus formation in AF are disease specific or also evident within the normal heart. The present study examined whether there are differences in, platelet reactivity, endothelial function and inflammation in blood samples obtained from intracardiac and peripheral sites in subjects within structurally normal hearts. Sixteen patients with diagnosed left sided supraventricular tachycardia (SVT) undergoing a routine elective electrophysiological study and ablation were studied. Blood samples were taken simultaneously from the femoral vein, right atrium and left atrium, immediately following transseptal puncture and prior to heparin bolus administration. Between peripheral and atrial sample sites patients with SVT showed no change in platelet reactivity or aggregation (CD62P P=0.91, sCD40L P=0.9), thrombus formation (TAT P=0.55), endothelial function (vWF P= 0.75, ADMA P=0.97 and NO P=0.61), or inflammation (VCAM‐1 p=0.59 and ICAM‐1 p=0.69). SVT patients had lower ADMA and ICAM‐1 levels than AF patients. This study demonstrates for the first time in SVT subjects with structurally normal hearts have consistent haemostatic function between atrial and peripheral sites. These results suggest that the atria of SVT patients do not contain predisposing thrombogenic, endothelial of inflammatory factors that promote/initiate thrombus formation. This article is protected by copyright. All rights reserved.
    May 14, 2014   doi: 10.1111/1440-1681.12256   open full text
  • αB‐Crystallin R120G variant causes cardiac arrhythmias and alterations in the expression of Ca2+ handling proteins and ER stress in mice.
    Qibin Jiao, Atsushi Sanbe, Xingwei Zhang, Jun‐Ping Liu, Susumu Minamisawa.
    Clinical and Experimental Pharmacology and Physiology. May 13, 2014
    Mutations of αB‐crystallin (CryαB), a small heat shock protein abundantly expressed in cardiac and skeletal muscles, are known to cause desmin‐related myopathies. The CryαB R120G allele has been linked to a familial desminopathy, and in the transgenic mice, causes a sudden death at about 28 weeks of age. To investigate the mechanisms of the sudden cardiac arrest of the CryαB R120G transgenic mice, we prepared the protein samples from the left ventricular tissues of the two different age groups (10 verses 28 weeks), and examined Ca2+ handling proteins. We found that the expression levels of SERCA2,phospholamban, ryanodine receptor 2, and calsequestrin 2 were significantly decreased in the 28‐week old CryαB R120G transgenic mice. In addition, low heart rate variability (HRV) such as heart rate, total power and low frequency was observed,and continuous ECG monitoring uncovered cardiac arrhythmias such as ventricular tachycardia, atrioventricular block and atrial flutter in the 28‐week old CryαB R120G transgenic mice. In contrast, we found that the expression levels of endoplasmic reticulum (ER) degradation enhancing, α‐mannosidase‐like protein (EDEM), inositol requirement 1 (IRE1) and x‐box binding protein 1 (XBP1) were significantly increased in the 28‐week old CryαBR120G transgenic mice, suggesting that the CryαBR120G transgenic mice exhibited an increased ER stress. Together, our data suggest that the CryαB R120G dominant variant induces ER stress and impairs Ca2+ regulation leading to an aging related cardiac dysfunction, arrhythmias and decreased autonomous tone with shortened lifespan. This article is protected by copyright. All rights reserved.
    May 13, 2014   doi: 10.1111/1440-1681.12253   open full text
  • Molecular mechanisms of aging and related diseases.
    Jun‐Ping Liu.
    Clinical and Experimental Pharmacology and Physiology. May 06, 2014
    Human and other multi‐cellular life species age and the aging processes become dominant in a late phase of lives. However, recent studies challenged the dogma suggesting that ageing in some animal species may never occur, but mammals undertake cell replicative senescence as early as before birth in embryos under physiological conditions. How the molecular machineries operate, and why aging cells dominate under some circumstances, are intriguing. Recent studies show that cell aging involves extensive cellular remodeling, including telomere attrition, heterochromatin formation, endoplasmic reticulum stress, mitochondrial disorders, lysosome processing organelles and chromatins. This article provides an update in the molecular mechanisms underlying aging of various cell types, the newly described developmental and programmed replicative senescence, and the critical roles of cellular organelles and effectors in Parkinson's disease, diabetes, hypertension and dyskeratosis congenita. This article is protected by copyright. All rights reserved.
    May 06, 2014   doi: 10.1111/1440-1681.12247   open full text
  • Involvement of Imperatorin Derivative‐OW1 in the antihypertensive effect and vascular remodeling of renovascular hypertension rats.
    Nan Zhou, Tao Wang, Jia Song, Huaizhen He, Jianyu He, Langchong He.
    Clinical and Experimental Pharmacology and Physiology. May 06, 2014
    OW1 is a novel imperatorin derivative that exhibits vasodilative activity. In the present study the antihypertensive effect and inhibition effect on vascular remodeling of OW1 were investigated in 2K1C renovascular hypertensive rats. OW1‐induced vasodilatation in rat mesenteric arteries and renal arteries was evaluated in vitro. 2K1C renovascular hypertensive rats were developed and treated with OW1 (40 and 80 mg/kg/day) for 8 weeks. Blood pressure of rats was measured in conscious. Concentration of microalbumin (mALB) and total protein (U‐TP) of urine and the levels of angiotensin II (Ang II), calcitonin gene‐related peptide (CGRP), and angiotensin‐converting enzyme 1 (ACE) of plasma were detected. The unclipped kidney was stained with hematoxylin and eosin (H&E) and Masson trichrome. The sectioned aortas were stained with Masson trichrome. Immunohistochemical analysis was used to quantify the collagen I and III. OW1 relaxed the rat mesenteric and renal arterial rings in vitro. Treatment of 2K1C hypertensive rats with OW1 (40 and 80 mg/kg/day) for 8 weeks significantly decreased blood pressure Besides, OW1 reduced the Ang II and ACE concentration and raised the CGRP concentration in rat plasma. High doses of OW1 decreased the levels of blood urea nitrogen, mALB and U‐TP. Histology results demonstrated that OW1 reduced renal arteriolar thickness and relieved the structural hypertrophic arteries. Moreover, OW1 caused an inhibition effect on vascular remodeling and renal lesions in hypertensive rats. In summary, the results suggested that OW1 could be a potential novel candidate for hypertension intervention. This article is protected by copyright. All rights reserved.
    May 06, 2014   doi: 10.1111/1440-1681.12248   open full text
  • Anti‐hypertensive and anti‐inflammatory actions of combined azilsartan and chlorthalidone in Dahl salt‐sensitive rats on a high‐fat, high‐salt diet.
    Chunhua Jin, Sean O'Boyle, Daniel T. Kleven, Jennifer S. Pollock, David M. Pollock, John J. White.
    Clinical and Experimental Pharmacology and Physiology. May 06, 2014
    The metabolic syndrome (MetS) and chronic kidney disease are global health issues. The MetS induces hypertension (HTN) and commonly results in renal damage. The optimal therapy for HTN in the MetS is unknown. Thiazide diuretics are first line therapy; however, these drugs may have untoward effects. This study investigated the effects of azilsartan (AZL), chlorthalidone (CLTD), and the combination (AZL + CLTD) on blood pressure and renal injury in a rodent model with features of the MetS. Dahl salt‐sensitive (Dahl S) rats were fed high‐fat (36% fat) high‐salt (4% NaCl) diet. Groups were then treated with vehicle, AZL (3 mg/kg/day), CLTD (5 mg/kg/day) or AZL+CLTD. Mean arterial pressure (MAP) was recorded continuously by telemetry. After 26 days, rats were humanely killed and kidneys were harvested for histology. Both AZL and CLTD attenuated the rise in blood pressure compared to vehicle, and the combination further reduced blood pressure compared to CLTD alone. All treatments reduced proteinuria and albuminuria. Only groups treated with AZL prevented nephrinuria. Nephrinuria was 57% lower, and proteinuria was 47% lower with combination therapy compared to AZL alone. All treatments reduced the number of inflammatory cells in the kidney. In conclusion, in our model, AZL and CLTD lower blood pressure and exhibit renal protective effects. AZL treatment offers additional protection as evidenced by lower nephrinuria and plasma MCP‐1. Combination therapy afforded the greatest protective effects and may be the best choice for hypertensive therapy in the MetS. This article is protected by copyright. All rights reserved.
    May 06, 2014   doi: 10.1111/1440-1681.12250   open full text
  • Natriuretic peptide clearance receptor ligand (C‐ANP4‐23) attenuates angiogenesis in a murine sponge implant model.
    S.A. Almeida, C.C. Cardoso, L.A.A. Orellano, A.M. Reis, L.S. Barcelos, S.P. Andrade.
    Clinical and Experimental Pharmacology and Physiology. May 06, 2014
    Natriuretic peptide receptor‐C (NPR‐C) activation by C‐ANP4‐23, a specific agonist for this receptor, has been shown to inhibit key events of the angiogenic cascade (migration, proliferation, and vascular endothelial growth factor—VEGF—production). We investigated whether C‐ANP4‐23 could also inhibit angiogenesis in the sponge model in vivo. In this study, we evaluated the effects of this peptide on inflammatory and angiogenic components of the fibrovascular tissue induced by polyether polyurethane sponge implants in mice. The hemoglobin content (μg/mg wet tissue) and blood flow measurements (laser Doppler perfusion imaging) of the implants, used as an index of vascularization, showed that single or multiple doses of C‐ANP4‐23 reduced angiogenesis in the implants relative to the PBS‐treated group. The peptide exerted an inhibitory effect on nitric oxide production (nitrite levels) and a dual effect on VEGF levels, depending on the number of doses. Histological analysis corroborated the biochemical and functional parameters indicative of neovascularization inhibition (decreased vessel number). The peptide failed to modulate inflammation in our system. The inhibitory effect of C‐ANP4‐23 on the angiogenic component of the fibrovascular tissue induced by the synthetic matrix extends the range of the peptide's actions and may indicate its therapeutic potential in controlling angiogenesis in fibroproliferative diseases. This article is protected by copyright. All rights reserved.
    May 06, 2014   doi: 10.1111/1440-1681.12251   open full text
  • Asymmetric dimethyl arginine does not inhibit arginase activity and is pro‐proliferative in pulmonary endothelial cells.
    Bernadette Chen, Krista Strauch, Yi Jin, Hongmei Cui, Leif D Nelin, Louis G Chicoine.
    Clinical and Experimental Pharmacology and Physiology. May 06, 2014
    Asymmetric dimethyl arginine (ADMA) is an endogenously produced nitric oxide synthase (NOS) inhibitor. L‐arginine can be metabolized by NOS and arginase, and arginase is the first step in polyamine production necessary for cellular proliferation. We tested the hypothesis that ADMA would inhibit NOS but not arginase activity, and that this pattern of inhibition would result in greater L‐arginine bioavailability to arginase and thereby increase viable cell number. Bovine arginase was used in in vitro activity assays with various concentrations of substrate (L‐arginine, ADMA, NG‐monomethyl‐L‐arginine (L‐NMMA) and NG‐nitro‐L‐arginine methyl ester (L‐NAME)). Only L‐arginine resulted in measurable urea production (Km = 6.9 ± 0.8 mM; Vmax = 6.6 ± 0.3 μmol/mg protein/min). We then incubated bovine arginase with increasing concentrations of ADMA, L‐NMMA, and L‐NAME in the presence of 1 mM L‐arginine, and found no effect of any of the tested compounds on arginase activity. Using bovine pulmonary arterial endothelial cells (bPAEC) we determined the effects of ADMA on NO and urea production, and found a significantly lower NO production and greater urea production (p<0.003) with ADMA, without changes in arginase protein levels. Additionally, ADMA treatment resulted in ~30% greater number of viable cells after 48 hours than in control bPAEC. These results demonstrate that ADMA is neither a substrate nor an inhibitor of arginase activity, and that in bPAEC ADMA inhibits NO production and enhances urea production leading to more viable cells. These results may have pathophysiological implications in disorders associated with higher ADMA levels such as pulmonary hypertension. This article is protected by copyright. All rights reserved.
    May 06, 2014   doi: 10.1111/1440-1681.12252   open full text
  • Effects of entecavir on peripheral blood lymphocyte profiles in chronic hepatitis B patients with suboptimal responses to adefovir.
    Liwen Zhang, Quan Wang, Xiaoli Hu, Pingwei Zhao, Yanfang Jiang.
    Clinical and Experimental Pharmacology and Physiology. April 29, 2014
    The aim of this study was to assess the long term impact of ETV on T, B and NK cell immunity in patients with suboptimal responses to adefovir (SRA) chronic hepatitis B (CHB). Thirty SAR CHB patients and 20 age‐/gender‐matched healthy controls (HC) completed at least six months of ETV treatment. HBV DNA loads, ALT and AST, and the frequency of different subsets of T, B and NK cells in individual subjects were measured. There were smaller numbers of CD3‐CD56+ and CD244+ NK cells, CD3+CD8+ T cells, and cytokine‐secreting CD4+T cells, but greater numbers of CD3+CD4+, CD4+CD25+Foxp3+, CD4+CD25+CD127low T cells and CD19+CD27+ B cells detected in SRA patients. After switching to ETV monotherapy, the levels of HBV DNA and HBsAg, as well as HBeAg seropositivity gradually decreased, accompanied by decreased levels of ALT and AST. Furthermore, the numbers of NK, CD8+ and cytokine‐secreting CD4+ T cells were increased, but CD4+, CD4+CD25+Foxp3+, CD4+CD25+CD127low T cells and CD19+CD27+ B cells in SRA CHB patients were decreased. The frequency of CD4+IFNγ+ T cells were negatively associated with serum HBV DNA levels. Thus, treatment with ETV inhibits HBV replication, modulates T and NK cell immunity and improves the liver function in SAR CHB patients. This article is protected by copyright. All rights reserved.
    April 29, 2014   doi: 10.1111/1440-1681.12245   open full text
  • Understanding the placebo effect in clinical trials for postural tachycardia syndrome.
    Victor C Nwazue, Amy C Arnold, Vidya Raj, Bonnie K Black, Italo Biaggioni, Sachin Y Paranjape, Carlos Orozco, William D Dupont, David Robertson, Satish R Raj.
    Clinical and Experimental Pharmacology and Physiology. April 23, 2014
    Postural tachycardia syndrome (POTS) is characterized by excessive increases in heart rate (HR) upon standing. Previous studies have shown that standing HR decreases over time in POTS patients given placebo. We hypothesized that this reduction is due to cardiovascular physiological alteration, as opposed to psychological benefit from perceived therapy. To prospectively test this hypothesis, we examined the effects of an open‐label ‘no treatment’ intervention (NoRx) compared with a patient‐blinded placebo on standing HR in POTS patients. Twenty‐one POTS patients participated in a randomized cross‐over trial with oral placebo versus NoRx administered at 0900 h. Seated blood pressure (BP) and HR were measured at baseline and every hour for 4 h. Similarly, BP and HR were measured while patients stood for 10 min at these time points. Standing HR decreased significantly over time with both NoRx (112 ± 13 and 103 ± 16 b.p.m. at baseline and 4 h, respectively) and placebo (112 ± 14 and 102 ± 16 b.p.m. at baseline and 4 h, respectively; Ptime < 0.001), but this effect was not different between interventions (Pdrug = 0.771). Postural tachycardia syndrome patients have exaggerated orthostatic tachycardia in the morning that decreases over time with either placebo or NoRx interventions, suggesting this phenomenon is due to cardiovascular physiological variation. These data highlight the need for a placebo arm in haemodynamic clinical trials in POTS and may have important implications for the diagnosis of these patients.
    April 23, 2014   doi: 10.1111/1440-1681.12221   open full text
  • GABAA receptors are involved in the analgesic effects of morphine microinjected into the central nucleus of the amygdala.
    Mina Rashvand, Ali Khajavai, Mohsen Parviz, Parisa Hasanein, Mansoor Keshavarz.
    Clinical and Experimental Pharmacology and Physiology. April 23, 2014
    The central nucleus of the amygdala (CeA) has an important role in pain perception and analgesia. Opioid and GABAA receptors, which are both involved in pain modulation, are found in high concentration in the CeA. The present study was designed to examine the interaction of opioidergic and GABAergic systems in the CeA during modulation of acute thermal pain. In the present study, different doses of morphine (25, 50 and 100 μg/rat), either alone or after 5 min pretreatment with the selective GABAA receptor agonist muscimol (60 ng/rat) or the selective GABAA receptor antagonist bicuculline (50 ng/rat), were injected bilaterally into the CeA of each rat. Tail‐flick latencies (TFL) were measured every 5 min for 60 min. The results revealed that microinjection of morphine into the CeA significantly increased TFL in a dose‐dependent manner. Microinjection of bicuculline or muscimol in combination with morphine into the CeA increased and decreased TFL, respectively. It seems that morphine in the CeA facilitates the function of descending inhibitory systems by interacting with the activity of local GABAA receptors.
    April 23, 2014   doi: 10.1111/1440-1681.12223   open full text
  • Stimulation of 5‐HT4 receptor enhances differentiation of mouse induced pluripotent stem cells into neural progenitor cells.
    Toshiaki Ishizuka, Hazuki Goshima, Ayako Ozawa, Yasuhiro Watanabe.
    Clinical and Experimental Pharmacology and Physiology. April 23, 2014
    Activation of serotonin (5‐hydroxytryptamine; 5‐HT) receptors plays a role in adult neurogenesis and differentiation of neural progenitor cells (NPC). Herein, we examined the involvement of 5‐HT receptors in the differentiation of mouse induced pluripotent stem (iPS) cells into NPC. To induce embryoid body (EB) formation, mouse iPS cells were cultured on ultralow‐attachment dishes. All‐trans retinoic acid (ATRA; 1 μmol/L) and/or 5‐HT (0.03 or 0.1 μmol/L) was added to the EB cultures for 4 days and then EB plated on gelatin‐coated plates were cultured for 7 or 14 days. Immunofluorescence staining revealed that mouse iPS cells expressed both 5‐HT2A and 5‐HT4 receptors and, to a lesser extent, 5‐HT1A receptors. Treatment with 5‐HT significantly enhanced the ATRA‐induced expression of nestin, a specific marker for NPC, and phosphorylation of cAMP response element‐binding protein (CREB). Pretreatment of EB cultures with either 1 μmol/L GR113808 (a selective 5‐HT4 receptor antagonist) or 1 μmol/L H89 (a protein kinase (PKA) inhibitor) significantly inhibited these effects of 5‐HT. These findings suggest that stimulation of 5‐HT4 receptors may enhance ATRA‐induced neural differentiation of mouse iPS cells through activation of PKA and CREB.
    April 23, 2014   doi: 10.1111/1440-1681.12224   open full text
  • Maternal fructose intake during pregnancy modulates hepatic and hypothalamic AMP‐activated protein kinase signalling in a sex‐specific manner in offspring.
    Yuuka Mukai, Haruka Ozaki, Yuko Serita, Shin Sato.
    Clinical and Experimental Pharmacology and Physiology. April 23, 2014
    Dietary fructose ingestion during gestation affects carbohydrate metabolism in the offspring. In the present study, we investigated the effects of excess fructose intake during pregnancy on hepatic and hypothalamic AMP‐activated kinase (AMPK) expression and phosphorylation, as well as hepatic glucose‐6‐phosphatase (G6Pase) activity in offspring. Pregnant Wistar rats received normal chow and 100 g/L fructose solution or normal water during gestation ad libitum. On gestational Day 21, some dams were killed and plasma samples and fetuses were collected. The remaining dams received normal water after spontaneous delivery during lactation. Pups were killed on postnatal Day 22 and the plasma, liver and hypothalamus were collected and analysed. Plasma glucose and insulin levels increased in female but not male offspring in the fructose group. Although the mRNA and total protein levels of AMPKα were unchanged, levels of phosphorylated AMPKα protein in the fructose group of female offspring were significantly lower in the liver and 4.6‐fold higher in the hypothalamus. The hepatic protein level of sirtuin 1, which is involved in AMPK phosphorylation and activation, was significantly reduced in the fructose group of female offspring. The activity of G6Pase, which plays a role in gluconeogenesis, was significantly enhanced in the liver of female offspring from fructose‐fed dams. These changes were not observed in male offspring. In conclusion, we found that excessively high fructose intake during pregnancy may modulate the hepatic and hypothalamic AMPK signalling pathways in female offspring after birth.
    April 23, 2014   doi: 10.1111/1440-1681.12225   open full text
  • MicroRNA‐130a regulates autophagy of endothelial progenitor cells through Runx3.
    Quanfu Xu, Shu Meng, Bo Liu, Mao‐Quan Li, Yeting Li, Lu Fang, Yi‐Gang Li.
    Clinical and Experimental Pharmacology and Physiology. April 23, 2014
    Dysfunction of endothelial progenitor cells (EPC) contribute to diabetic vascular disease. MicroRNAs (miRNAs) are key regulators of diverse cellular processes, including angiogenesis. We recently reported that downregulated miR‐130a in patients with Type 2 diabetes mellitus (DM) results in EPC dysfunction, including increased apoptosis, likely via its target runt‐related transcription factor 3 (Runx3). However, whether miR‐130a affects the autophagy of EPC is unknown. The aim of the present study was to explore the effects of miR‐130a on the autophagy and cell death of EPC, as well as their expression of Beclin 1 (BECN1; an initiator of autophagosome formation) and the anti‐apoptotic protein Bcl2 (which binds to and inactivates BECN1), and the role of Runx3 in mediating these effects. The EPC were cultured from peripheral blood mononuclear cells of diabetic patients and non‐diabetic controls. Cells were transfected with an miR‐130a inhibitor, or mimic‐miR‐130a or mimic‐miR‐130a plus lentiviral vector expressing Runx3 to manipulate miR‐130a and/or Runx3 levels. The number of autophagosomes was counted under transmission electron microscopy and cell death was examined by flow cytometry. The mRNA expression of Beclin1 was measured by real‐time polymerase chain reaction and the protein expression of Beclin1 and Bcl2 was determined by western blotting. Both the number of autophagosomes and Beclin1 expression were increased in EPC from patients with DM. Inhibition of miR‐130a increased the number of autophagosomes and Beclin1 expression, but attenuated Bcl2 expression. Overexpression of miR‐130a decreased the number of autophagosomes, cell death and Beclin1 expression, but promoted Bcl2 expression; these effects were mediated by Runx3. In conclusion, miR‐130a is important for maintaining normal autophagy levels and promoting the survival of EPC via regulation of Bcl‐2 and Beclin1 expression, via Runx3. MiR‐130a may be a regulator linking apoptosis and the autophagy of EPC.
    April 23, 2014   doi: 10.1111/1440-1681.12227   open full text
  • Assessment of various second‐line medications in addition to inhaled corticosteroid in asthmatic patients ‐ A randomized controlled trial.
    Rajanandh MG, Nageswari AD, Ilango K.
    Clinical and Experimental Pharmacology and Physiology. April 17, 2014
    Many patients with persistent asthma could not achieve the treatment goal for asthma with a single controller medication. The study aimed to assess the lung function and rescue medication usage in asthmatic patients receiving four different categories of drugs in combination with an inhaled corticosteroid. Recruited patients were randomized into four groups to receive Budesonide with Formoterol, Doxofylline, Montelukast and Tiotropium for a period of three months. Lung function i.e. Forced Expiratory Volume in one second (FEV1) and rescue medication usage was measured at baseline, day 15, 30, 45, 60 and 90. A total of 297 patients completed the study. At baseline, no significant difference (P>0.05) was observed in any of the outcome measures. Significant within‐group improvement in% FEV1 was observed in all the groups. At day 90, between‐group difference showed that improvement in% FEV1 was significantly (p<0.05) high for budesonide plus formoterol followed by budesonide plus montelukast, budesonide plus doxofylline and least for budesonide plus tiotropium. Similarly, within‐group and between‐group comparisons showed significant (p<0.05) reduction in rescue medication usage in all the groups. The intensity in decrease was more in budesonide plus formoterol group followed by budesonide plus montelukast, budesonide plus doxofylline and budesonide plus tiotropium groups. Based on our findings, among the second‐line treatment regimens, budesonide with either montelukast or doxofylline was found to be better than budesonide plus tiotropium in mild to moderate persistent asthmatic patients. Further studies with a longer duration are likely to be useful. This article is protected by copyright. All rights reserved.
    April 17, 2014   doi: 10.1111/1440-1681.12239   open full text
  • TNF‐α Inhibition with Lenalidomide Alleviates Tissue Oxidative Injury and Apoptosis in ob/ob Obese mice.
    Xiaoling Zhu, Shasha Jiang, Nan Hu, Fuling Luo, Hailong Dong, Yu‐Ming Kang, Kyla R. Jones, Yunzeng Zou, Lize Xiong, Jun Ren.
    Clinical and Experimental Pharmacology and Physiology. April 17, 2014
    Lenalidomide (Revlimid), an analogue of thalidomide, possesses potent cytokine modulatory capacity through inhibition of cytokines such as TNF‐α, a cytokine pivotal for the onset and development of complications in obesity and diabetes mellitus. This study was designed to evaluate the effect of lenalidomide on oxidative stress, protein and DNA damage in multiple organs in an ob/ob murine model of obesity. C57BL/6 lean and ob/ob obese mice were given lenalidomide (50 mg/kg/d, p.o.) for 5 days. Oxidative stress, protein and DNA damage were assessed using glutathione (GSH) to oxidized glutathione (GSSG) conversion, carbonyl formation and Comet assay, respectively. Apoptosis was evaluated using caspase‐3 activity and levels of Bax, Bcl‐2, Bip, Caspase‐8, Caspase‐9 and TNF‐α were assessed using western blot analysis. Our data revealed that lenalidomide treatment did not affect glucose clearance in lean or ob/ob mice. Obese mice displayed reduced GSH/GSSG ratio in liver, gastrocnemius skeletal muscle and small intestinal tissues, enhanced protein carbonyl formation, DNA damage and caspase‐3 activity in liver, kidney, skeletal muscle and intestines tissues, the effects of which were alleviated by lenalidomide with the exception of obesity‐associated DNA damage in liver and kidney. Western blot analysis revealed elevated TNF‐α, Bax, Bcl‐2, Bip, Caspase‐8, and Caspase‐9 in ob/ob mice with various degrees of reversals by lenalidomide treatment. Taken together, these data indicated that lenalidomide protects against obesity‐induced tissue injury and protein damage possibly associated with antagonism of cytokine production and cytokine‐induced apoptosis and oxidative stress. This article is protected by copyright. All rights reserved.
    April 17, 2014   doi: 10.1111/1440-1681.12240   open full text
  • The toll‐like receptor‐4 signalling in the progression of non‐alcoholic fatty liver disease induced by high fat and high fructose diet in mice.
    Jing Liu, Zhen‐jie Zhuang, Dong‐xue Bian, Xiao‐jie Ma, Yun‐hao Xun, Wen‐jun Yang, Yan Luo, Yin‐lan Liu, Ling Jia, Yan Wang, Ming‐li Zhu, De‐wei Ye, Gang Zhou, Guo‐qiang Lou, Jun‐ping Shi.
    Clinical and Experimental Pharmacology and Physiology. April 17, 2014
    The aim of the present study was to investigate toll‐like receptor‐4 (TLR4) signalling at different stages of non‐alcoholic fatty liver disease (NAFLD) induced by high fat high fructose (HFHFr) diet in mice. TLR4 wild type (WT) and mutant (TLR4mut) mice were fed with either standard chow (SC) or HFHFr diet for different periods of time from 4 to 16 weeks. The liver pathological characteristics and functions were assessed. Simple steatosis, steatohepatitis and hepatic fibrosis occurred sequentially in week 4, 8 and 16 in the WT mice fed with HFHFr. The expression levels of TLR4, MyD88, IRF3 and IRF7 started to increase at the time course of week 4, peaked at week 8, and then declined to basal levels at week 16. This alteration pattern was consistent with the changes of inflammation in the livers revealed by H&E staining. However, lipid accumulation, inflammation and fibrosis in the livers of TLR4mut mice given HFHFr diet were significantly alleviated. In addition, the expression of activin‐A in TLR4‐WT mice fed with HFHFr diet increased at week 16. Our dada suggest that TLR4 signaling mediates non‐alcoholic steatohepatitis before fibrosis, and activin‐A is subsequently involved in NAFLD. This article is protected by copyright. All rights reserved.
    April 17, 2014   doi: 10.1111/1440-1681.12241   open full text
  • Antagonism of Salvianolic acid B on LPS‐induced disseminated intravascular coagulation in rabbits.
    Zheng Wu, Jian‐nan Li, Zhi‐quan Bai, Xi Lin.
    Clinical and Experimental Pharmacology and Physiology. April 17, 2014
    Aim To study the effect of Salvianolic acid B (Sal B) on lipopolysaccharide (LPS)‐induced disseminated intravascular coagulation (DIC) in rabbits. Methods Continuous infusion of LPS induced DIC model in rabbits. The treatments with Salvianolic acid B were started simultaneously with LPS infusion through the contralateral marginal ear vein. activated partial thromboplastin time (APTT), prothrombin time (PT), platelet count and fibrinogen concentration were measured; the plasma levels of fibrin‐fibrinogen degradation products (FDP) were detected; alanine aminotransferase (ALT) and blood urine nitrogen (BUN), the activity of protein C, antithrombinIII (ATIII) and the concentration of TNF‐α were determined. Results The gradual impairment of hemostatic parameters was induced by continuous infusion of LPS. The APTT, PT, BUN, ALT and plasma TNF‐α increased obviously, however, the platelet count, fibrinogen, FDP, protein C and ATIII decreased clearly. The intravenous administration of Salvianolic acid B attenuated the increased levels of APTT, PT, BUN, ALT and plasma TNF‐α, and the decreased levels of fibrinogen, platelet, FDP, protein C and ATIII induced by LPS infusion. Conclusion Salvianolic acid B has the effect against LPS induced DIC in rabbits This article is protected by copyright. All rights reserved.
    April 17, 2014   doi: 10.1111/1440-1681.12242   open full text
  • Impact of short‐term low‐dose atorvastatin on LDL and HDL subfraction phenotype.
    Rui‐Xia Xu, Yuan‐Lin Guo, Xiao‐Lin Li, Sha Li, Jian‐Jun Li.
    Clinical and Experimental Pharmacology and Physiology. April 17, 2014
    Statins can significantly reduce low density lipoprotein (LDL) cholesterol (LDL‐C) and modestly raise or not alter high density lipoprotein (HDL) cholesterol (HDL‐C). However, their impact on HDL subfractions and LDL subfractions has less been examined. The objective of this study was to investigate the short‐term impact of low‐dose atorvastatin on HDL and LDL subfractions in humans. In this randomized study, data from 52 subjects were analyzed. Of them, 37 patients with atherosclerosis were randomized to atorvastatin 10mg/d (n=17) or 20 mg/d (n=20) treatment and 15 healthy subjects without therapy were used as controls for 8 weeks. The lipid profile and lipoprotein subfractions were determined by Lipoprint system at baseline and 8 weeks. Data suggested that atorvastatin treatment (10 mg/d and 20 mg/d) for 8 weeks significantly decreased LDL‐C levels and reduced the cholesterol concentration of all LDL subfractions accompanied by an increase of the mean LDL particle size. Although atorvastatin 10 mg/d treatment for 8 weeks had no any impact on HDL subfraction atorvastatin 20 mg/d treatment for 8 weeks significantly increased the cholesterol concentration of large HDL particles and decreased the cholesterol concentration of small HDL particles without changes of serum HDL‐C level in patients with atherosclerosis. Therefore, our results suggested that atorvastatin 20 mg/d treatment for 8 weeks could result in a favorable modification of HDL subfraction phenotype besides its impacts on cholesterol concentration of all LDL subfractions and mean LDL particle size. This article is protected by copyright. All rights reserved.
    April 17, 2014   doi: 10.1111/1440-1681.12243   open full text
  • Alpha‐lipoic acid prolongs survival and attenuates acute kidney injury in a rat model of sepsis.
    Guofu Li, Linlin Gao, Jia Jia, Xiaoying Gong, Bin Zang, Weimin Chen.
    Clinical and Experimental Pharmacology and Physiology. April 17, 2014
    Acute kidney injury is a frequent and serious complication in patients with severe sepsis. Alpha‐lipoic acid (ALA), a naturally occurring dithiol compound, has been shown to possess anti‐inflammatory and anti‐oxidative properties. In the present study, we investigated whether ALA can attenuate acute kidney injury and improve survival in a rat model of sepsis. Rats were subjected to cecal ligation and puncture (CLP) surgery to induce sepsis. ALA (200 mg/kg) was administrated by oral gavage immediately (early treatment) or 12 h (delayed treatment) after the surgical procedure. The results showed that both early and delayed treatment with ALA effectively prolonged survival, improved pathological damages in kidney tissues, and reduced serum blood urea nitrogen (BUN) and creatinine (Cr) levels in CLP‐induced septic rats. Furthermore, early treatment with ALA dramatically inhibited the release of tumor necrosis factor (TNF)‐α, interleukin (IL)‐6 and IL‐1β in serum and reduced the mRNA and protein expression levels of inducible nitric oxide synthase (iNOS) and high mobility group box‐1 (HMGB‐1) in kidney tissues from CLP‐induced rats. Finally, CLP‐induced nuclear factor (NF)‐κB activation in kidney tissues was also significantly suppressed by early treatment with ALA. Taken together, we conclude that ALA is able to reduce mortality and attenuate acute kidney injury associated with sepsis, possibly by the anti‐inflammatory actions. ALA may be a promising novel agent for the treatment of conditions associated with septic shock. This article is protected by copyright. All rights reserved.
    April 17, 2014   doi: 10.1111/1440-1681.12244   open full text
  • Exogenous L‐arginine attenuates the effects of angiotensin II on renal hemodynamics and the pressure natriuresis–diuresis relationship.
    Satarupa Das, David L Mattson.
    Clinical and Experimental Pharmacology and Physiology. March 25, 2014
    Administration of exogenous L‐arginine (L‐Arg) attenuates angiotensin‐II (AngII)‐mediated hypertension and kidney disease in rats. The present study assessed renal hemodynamics and pressure diuresis–natriuresis in anaesthetized rats infused with vehicle, AngII (20 ng/kg per min i.v.) or AngII + L‐Arg (300 μg/kg per min i.v.). Experiments in isolated aortic rings were carried out to assess L‐Arg effects on the vasculature. Increasing renal perfusion pressure (RPP) from ˜100 to 140 mmHg resulted in a nine‐ to tenfold increase in urine flow and sodium excretion rate in control animals. In comparison, AngII infusion significantly reduced renal blood flow (RBF) and glomerular filtration rate (GFR) by 40–42%, and blunted the pressure‐dependent increase in urine flow and sodium excretion rate by 54–58% at elevated RPP. Supplementation of L‐Arg reversed the vasoconstrictor effects of AngII and restored pressure‐dependent diuresis to levels not significantly different from control rats. Dose‐dependent contraction to AngII (10−10 mol/L to 10−7 mol/L) was observed with a maximal force equal to 27 ± 3% of the response to 10−5 mol/L phenylephrine. Contraction to 10−7 mol/L AngII was blunted by 75 ± 3% with 10−4 mol/L L‐Arg. The influence of L‐Arg to blunt AngII‐mediated contraction was eliminated by endothelial denudation or incubation with nitric oxide synthase inhibitors. Furthermore, the addition of 10−3 mol/L cationic or neutral amino acids, which compete with L‐Arg for cellular uptake, blocked the effect of L‐Arg. Anionic amino acids did not influence the effects of L‐Arg on AngII‐mediated contraction. These studies show that L‐Arg blunts AngII‐mediated vascular contraction by an endothelial‐ and nitric oxide synthase‐dependent mechanism involving cellular uptake of L‐Arg.
    March 25, 2014   doi: 10.1111/1440-1681.12212   open full text
  • Effects of opioid (tramadol) treatment on testicular functions in adult male rats: The role of nitric oxide and oxidative stress.
    Marwa A Ahmed, Adel Kurkar.
    Clinical and Experimental Pharmacology and Physiology. March 25, 2014
    Nowadays, tramadol hydrochloride is frequently used as a pain reliever, and for the treatment of premature ejaculation. Decreased semen quality was noted in chronic tramadol users. The present study aimed to elucidate the effects of tramadol on the testicular functions of adult male rats. A total of 40 albino adult male rats were divided into control and tramadol groups, with 20 rats for each group. Rats of the tramadol group were subcutaneously injected with 40 mg/kg three times per week for 8 weeks. The control group received normal saline 0.9%. Blood samples from each animal were obtained. Plasma levels of different biochemical substances were determined. Nitric oxide was measured in testicular tissue samples. Those samples together with epididymal tissue samples were processed for histopathological examination. Tramadol significantly reduced plasma levels of luteinizing hormone, follicle‐stimulating hormone, testosterone and total cholesterol, but elevated prolactin and estradiol levels compared with the control group. In addition, tramadol increased the testicular levels of nitric oxide and lipid peroxidation, and decreased the anti‐oxidant enzymes activities significantly compared with the control group. The tramadol group showed decreased sperm count and motility, and numbers of primary spermatocytes, rounded spermatid and Leydig cells. Immunohistochemical examinations showed that tramadol increased the expression of endothelial nitric oxide synthase in testicular tissues. The present study showed that tramadol treatment affects the testicular function of adult male rats, and these effects might be through the overproduction of nitric oxide and oxidative stress induced by this drug.
    March 25, 2014   doi: 10.1111/1440-1681.12213   open full text
  • Involvement of purinergic receptors and NOD‐like receptor‐family protein 3‐inflammasome pathway in the adenosine triphosphate‐induced cytokine release from macrophages.
    Thomas Gicquel, Tatiana Victoni, Alain Fautrel, Sacha Robert, Florence Gleonnec, Marie Guezingar, Isabelle Couillin, Véronique Catros, Elisabeth Boichot, Vincent Lagente.
    Clinical and Experimental Pharmacology and Physiology. March 25, 2014
    Adenosine triphosphate (ATP) has been described as a danger signal activating the NOD‐like receptor‐family protein 3 (NLRP3)‐inflammasome leading to the pro‐inflammatory cytokine, interleukin (IL)‐1β, release in the lung. The NLRP3‐inflammasome pathway has been previously described to be involved in experimental collagen deposition and the development of pulmonary fibrosis. The aim of the present study was to investigate the role of the NLRP3 inflammasome pathway and P2X7 purinergic receptor in the activation of human macrophages in vitro by ATP. We showed that adenosine 5′‐[γ‐thio]triphosphate tetralithium salt (ATPγS) and 2′,3′‐O‐(4‐benzoylbenzoyl) adenosine 5′‐triphosphate (BzATP), two stable analogs of ATP, are able to potentiate the release of IL‐1β from human monocyte‐derived macrophages induced by low concentration of lipopolysaccharide (LPS). However, in the same conditions no increase in IL‐1α and IL‐6 was observed. Immunochemistry has shown that human macrophages natively express NLRP3 and purinergic P2X7 receptors (P2X7R). NLRP3 and IL‐1β mRNA expression were induced from LPS‐primed macrophages, but also after 5‐h treatment of BzATP as analysed by reverse transcription quantitative polymerase chain reaction. However, other inflammasome pathways (NLRP1, NLRP2, NLRC4, NLRP6 and AIM2) and P2X7R were not induced by BzATP. We observed that P2X7R antagonists, A‐438079 and A‐740003, were able to reduce the release of IL‐1β, but not of IL‐1α and IL‐6 from macrophages stimulated by ATPγS or BzATP. The present results showed the involvement of the P2X7R‐NLRP3 inflammasome pathway in the secretion of IL‐1β from ATP‐stimulated human macrophages, and suggest that P2X7R were not involved in IL‐1α and IL‐6 release. This study also points out that repression of the P2X7R represents a novel potential therapeutic approach to control fibrosis in lung injury.
    March 25, 2014   doi: 10.1111/1440-1681.12214   open full text
  • Inhibition of acid‐sensing ion channel currents by propofol in rat dorsal root ganglion neurons.
    Zhen Lei, Xiaoyu Li, Guizhi Wang, Jianchun Fei, Tao Meng, Xinyu Zhang, Jingya Yu, Jingui Yu, Jingxin Li.
    Clinical and Experimental Pharmacology and Physiology. March 25, 2014
    Acid‐sensing ion channels (ASICs), part of the epithelial sodium channel/degenerin family, are activated by extracellular protons. The ASICs play a significant role in the acidosis‐mediated perception of pain. The anaesthetic agent propofol also exerts antinociceptive effects, but the underlying mechanisms for this effect are not clear. We used whole‐cell patch clamping to investigate the effect of propofol on proton‐gated currents in: (i) rat dorsal root ganglion (DRG) neurons; and (ii) HEK293 cells transfected with either ASIC1a or ASIC3. Propofol inhibited the amplitude of proton‐gated currents in DRG neurons, but did not change the sensitivity of ASICs to H+. Notably, propofol altered acid‐evoked excitability of rat DRG neurons and decreased the number of action potentials induced by acid stimuli. In addition, we demonstrated that propofol inhibited ASICs by directly binding with these channels in HEK293 cells. These results suggest that propofol inhibits proton‐gated currents in DRG neurons and that inhibition of proton‐gated currents explains, in part, the antinociceptive effects of propofol in primary afferent neurons.
    March 25, 2014   doi: 10.1111/1440-1681.12215   open full text
  • Melatonin and tadalafil treatment improves erectile dysfunction after spinal cord injury in rats.
    Hasan Hüseyin Tavukçu, Tarik Emre Şener, İlker Tinay, Cem Akbal, Mehmet Erşahin, Özge Çevik, Selin Çadırcı, Russel J Reiter, Göksel Şener.
    Clinical and Experimental Pharmacology and Physiology. March 25, 2014
    Oxidative stress plays an important role both in spinal cord injury (SCI) and erectile dysfunction (ED). The present study investigated the effects of melatonin and tadalafil treatment alone or in combination on SCI‐induced ED. Male Wistar albino rats (n = 40) were divided into five groups: sham‐operated control and SCI‐injured rats given either vehicle, melatonin (10 mg/kg, i.p.), tadalafil (10 mg/kg, p.o.) or a combination of melatonin and tadalafil. Spinal cord injury was induced using a standard weight‐drop method. On Day 7 after SCI, intracavernosal pressure (ICP) was measured and all rats were decapitated. Cavernosal tissues were obtained to examine caspase 3, nitric oxide synthase (NOS), myeloperoxidase (MPO) and superoxide dismutase (SOD) activities, as well as cGMP, nerve growth factor (NGF), malondialdehyde (MDA) and glutathione (GSH) levels. Spinal cord injury caused oxidative damage, as evidenced by increases in MDA and cGMP levels. In addition, MPO and caspase 3 activites were increased after SCI, whereas GSH and NGF levels and SOD activity were reduced. Melatonin effectively reversed these oxidative changes. Furthermore, in rats treated with both melatonin and tadalafil, the recoveries were more pronounced than in rats given either melatonin or tadalafil alone. The ICP/mean arterial pressure value in vehicle‐treated SCI rats was significantly higher than in the control group, whereas in the tadalafil‐ and tadalafil + melatonin‐treated groups have returned this value had returned to control levels. As an individual treatment, and especially when combined with tadalafil, a well‐known agent in the treatment of ED, melatonin prevented SCI‐induced oxidative damage to cavernosal tissues and restored ED, most likely due to its anti‐oxidant effects.
    March 25, 2014   doi: 10.1111/1440-1681.12216   open full text
  • Bitter tastants induce relaxation of rat thoracic aorta precontracted with high K+.
    Wen‐Bo Sai, Meng‐Fei Yu, Ming‐Yu Wei, Zhongju Lu, Yun‐Min Zheng, Yong‐Xiao Wang, Gangjian Qin, Donglin Guo, Guangju Ji, Jinhua Shen, Qing‐Hua Liu.
    Clinical and Experimental Pharmacology and Physiology. March 25, 2014
    It has been reported that bitter tastants decrease blood pressure and relax precontracted vascular smooth muscle. However, the underlying mechanisms remain unclear. The aim of the present study was to determine the mechanism underlying the vasorelaxant effect of the bitter tastants. Thoracic aortic rings were isolated from Wistar rats and contractions were measured using an isometric myograph. Intracellular Ca2+ ([Ca2+]i) in single rat thoracic aortic smooth muscle cells was recorded by calcium imaging. Calcium currents in single cells were recorded using patch‐clamp techniques. High K+ (140 mmol/L) induced contractions in rat thoracic aortic rings that were inhibited by 3 mmol/L chloroquine, 3 mmol/L denatonium and 10 μmol/L nifedipine. In single rat thoracic aortic smooth muscle cells, high K+ increased [Ca2+]i and this effect was also blocked by 3 mmol/L chloroquine and 10 μmol/L nifedipine. Under Ca2+‐free conditions, high K+ failed to induce contractions in rat thoracic aortic rings. On its own, chloroquine had no effect on the muscle tension of rat aortic rings and [Ca2+]i. The vasorelaxant effects of chloroquine on precontracted rat thoracic aortic rings were not altered by either 1 μg/mL pertussis toxin (PTX), an inhibitor of Gαo/i ‐protein, or 1 mmol/L gallein, an inhibitor of Gβγ‐protein. The results of patch‐clamp analysis in single cells indicate that 1 mmol/L chloroquine blocks voltage‐dependent L‐type Ca2+ channel (VDLCC) currents from both extracellular and intracellular sides. Together, the results indicate that chloroquine can block VDLCC, independent of PTX‐ and gallein‐sensitive G‐proteins, resulting in relaxation of high K+‐precontracted thoracic aortic smooth muscle.
    March 25, 2014   doi: 10.1111/1440-1681.12217   open full text
  • ATP‐binding cassette transporter A1 (ABCA1) promotes arsenic tolerance in human cells by reducing cellular arsenic accumulation.
    Xiaohua Tan, Li Yang, Lingling Xian, Jin Huang, Chunhong Di, Wenyi Gu, Shuli Guo, Lei Yang.
    Clinical and Experimental Pharmacology and Physiology. March 25, 2014
    Arsenic is a toxic element widely distributed in nature, such as water and soil. To survive this metalloid in the environment, nearly all organisms develop strategies to tolerate arsenic toxicity to some degree. Some arsenic‐resistance genes have been identified in bacteria and yeast, but for mammals, especially humans, these genes are largely unknown. The aim of the present study was to identify these genes and benefit our intervention of arsenic resistance. We first established a human arsenic‐resistant ECV‐304 (AsRE) cell line and then used suppression subtractive hybridization and microarray analysis to identify arsenic‐resistant genes in these cells. Of the significantly upregulated genes, three ATP‐binding cassette (ABC) subfamily members, namely ABCA1, ABCE1 and ABCF1, were chosen for further study with RNA interference and overexpression analyses. The 3‐(4,5‐dimethyl‐2 thiazoyl)‐2,5‐diphenyl‐2H‐tetrazolium bromide assay was used to determine the cell survival rate and the IC50, whereas atomic fluorescence spectrophotometry was used to determine intracellular arsenic levels. We found that among the three ABC genes, only when ABCA1 gene expression was silenced did cells obviously lose their arsenic tolerance. The arsenic accumulation in ABCA1 deficiency AsRE cells was greater than that in wild type AsRE cells. Overexpression of ABCA1 in HeLa cells decreased arsenic accumulation in the cells and the cells were more resistant to As(III) than control cells transfected with empty vector. These results suggest a new functional role for ABCA1 in the development of arsenic resistance in human cells.
    March 25, 2014   doi: 10.1111/1440-1681.12219   open full text
  • Antinociceptive and antidiarrheal effects of pioglitazone in a rat model of diarrhoea‐predominant irritable bowel syndrome: Role of nitric oxide.
    Pedram Paragomi, Reza Rahimian, Mohammad Hossein Kazemi, Mohammad Hadi Gharedaghi, Amin Khalifeh‐Soltani, Saeedeh Azary, Abbas Norouzi Javidan, Kamran Moradi, Stephen Sakuma, Ahmad Reza Dehpour.
    Clinical and Experimental Pharmacology and Physiology. January 28, 2014
    Irritable bowel syndrome (IBS) is a prevalent disease characterized by abdominal pain and abnormal bowel habits. Pioglitazone is a peroxisome proliferator‐activated receptor (PPAR) γ agonist and, although it is mostly used as an antidiabetic agent, it has been reported to have analgesic effects. Nitric oxide (NO), a gaseous molecule that mediates many of the effects of pioglitazone, has been implicated in the pathophysiology of IBS. The aim of the present study was to investigate the effects of pioglitazone on symptoms in a rat model of diarrhoea‐predominant IBS (D‐IBS).and to determine the role of NO in these effects. Diarrhoea‐predominant IBS was induced by intracolonic instillation of acetic acid. Pioglitazone (2 mg/kg, i.p.) was administered on Days 7, 9 and 11 after acetic acid instillation. To investigate the mechanism involved in pioglitazone action, rats were also administered either the PPARγ antagonist GW9662 (3 mg/kg, i.p.), the NO synthase (NOS) inhibitor NG‐nitro‐l‐arginine methyl ester (l‐NAME; 10 mg/kg, i.p.) or the NO precursor l‐arginine (250 mg/kg, i.p.) along with pioglitazone. Visceral hypersensitivity, nociceptive thresholds, defecation frequency, stool form, serum and colon NO production and inducible (i) NOS activity were assessed 1 h after the final injection of pioglitazone or dimethylsulphoxide (used as the vehicle). Pioglitazone reduced visceral hypersensitivity and defecation frequency, increased nociceptive thresholds, NO production and iNOS activity and shifted stool form towards hard stools in D‐IBS rats. These effects of pioglitazone were significantly reversed by l‐NAME, but not GW9662. l‐Arginine augmented the effects of pioglitazone. In conclusion, pioglitazone alleviates symptoms in a rat model of D‐IBS through an NO‐dependent mechanism.
    January 28, 2014   doi: 10.1111/1440-1681.12188   open full text
  • In vitro effects of cilostazol, a phosphodiesterase 3A inhibitor, on mouse oocyte maturation and morphology.
    Ahmed M Taiyeb, William L Dees, Mundhir T Ridha‐Albarzanchi, Christie M Sayes, Duane C Kraemer.
    Clinical and Experimental Pharmacology and Physiology. January 28, 2014
    Inhibition of phosphodiesterase 3A (PDE3A) in oocytes has been reported to arrest oocyte maturation and to increase intra‐oocyte cyclic adenosine monophosphate levels. Although many PDE3A inhibitors have been found to arrest oocyte maturation in different species, including humans, the most commonly prescribed PDE3A inhibitor named cilostazol (CLZ) has not yet been fully evaluated in reproduction. The present study was designed to investigate the potential inhibitory effects of CLZ on oocyte maturation and morphology in vitro. Antral oocytes were recovered from hyperstimulated mice and allocated to 10 different CLZ concentrations (0.00–67.66 μmol/L). Oocytes were then assessed after 24 and 48 h of incubation for maturation and morphology. Some of the evaluated CLZ concentrations (1.06–4.23 μmol/L) were made similar to those observed in human clinical trials. CLZ arrested oocyte maturation at the germinal vesicle (GV) stage at concentrations as low as 1.06 μmol/L (P < 0.0001). A selective degenerative impact of CLZ targeting arrested oocytes at the GV stage was observed during 24 h of incubation (r = −0.781, P < 0.0001). This was not the case with non‐arrested oocytes (r = −0.082, P = 0.64). Such degenerative impact was dose‐dependent (P < 0.0001), suggesting a role for cyclic adenosine monophosphate in this degenerative process. The degenerated oocytes were of distorted oolema or fragmented cytoplasm. Based on the experiments, it is concluded that CLZ can inhibit oocyte maturation in vitro, at concentrations similar to those observed in humans taking CLZ, and under such conditions the prolonged maintenance of oocytes at the GV stage is harmful.
    January 28, 2014   doi: 10.1111/1440-1681.12193   open full text
  • In vitro functional interactions of acetylcholine esterase inhibitors and muscarinic receptor antagonists in the urinary bladder of the rat.
    Uday K Killi, Vladimir Wsol, Ondrej Soukup, Kamil Kuca, Michael Winder, Gunnar Tobin.
    Clinical and Experimental Pharmacology and Physiology. January 28, 2014
    Obidoxime, a weak acetylcholine‐esterase (AChE) inhibitor, exerts muscarinic receptor antagonism with a significant muscarinic M2 receptor selective profile. The current examinations aimed to determine the functional significance of muscarinic M2 receptors in the state of AChE inhibition, elucidating muscarinic M2 and M3 receptor interaction. In the in vitro examinations, methacholine evoked concentration‐dependent bladder contractile and atrial frequency inhibitory responses. Although atropine abolished both, methoctramine (1 μmol/L) only affected the cholinergic response in the atrial preparations. However, in the presence of methoctramine, physostigmine, an AChE inhibitor, increased the basal tension of the bladder strip preparations (+68%), as well as the contractile responses to low concentrations of methacholine (< 5 μmol/L; +90–290%). In contrast to physostigmine, obidoxime alone raised the basal tension (+58%) and the responses to low concentrations of methacholine (< 5 μmol/L; +80–450%). Physostigmine concentration‐dependently increased methacholine‐evoked responses, similarly to obidoxime at low concentrations. However, at large concentrations (> 5 μmol/L), obidoxime, because of its unselective muscarinic receptor antagonism, inhibited the methacholine bladder responses. In conclusion, the current results show that muscarinic M2 receptors inhibit muscarinic M3 receptor‐evoked contractile responses to low concentrations of acetylcholine in the synaptic cleft. The muscarinic M2 and M3 receptor crosstalk could be a counteracting mechanism in the treatment of AChE inhibition when using reactivators, such as obidoxime.
    January 28, 2014   doi: 10.1111/1440-1681.12191   open full text
  • Advanced glycation end‐products impair Na+/K+‐ATPase activity in diabetic cardiomyopathy: Role of the adenosine monophosphate‐activated protein kinase/sirtuin 1 pathway.
    Qiong Yuan, Qian‐Yi Zhou, Du Liu, Lun Yu, Lin Zhan, Xiao‐Jing Li, Hong‐Yan Peng, Xiu‐Ling Zhang, Xin‐Chu Yuan.
    Clinical and Experimental Pharmacology and Physiology. January 28, 2014
    Decreased Na+/K+‐ATPase activity, and both sirtuin 1 (SIRT1) and adenosine monophosphate‐activated protein kinase (AMPK) have been reported to be involved in the development of diabetic cardiomyopathy (DCM). The present study aimed to investigate the advanced glycation end‐products (AGE) that impair Na+/K+‐ATPase stability by regulating the AMPK/SIRT1 pathway during progression of DCM. To study type 1 diabetic mellitus (T1DM), a disease model in rats was established by a single intraperitoneal injection of streptozotocin (STZ; 65 mg/kg), and neonatal rat cardiomyocytes were also cultured. Heart function was detected by Doppler, and SIRT1 and AMPK protein expression were detected by immunohistochemistry and western blotting. Na+/K+‐ATPase activity was also monitored. Using in vivo rat models of DCM, we showed that Na+/K+‐ATPase activity decreased when both AMPK and SIRT1 expression were downregulated. In vitro, AGE impaired Na+/K+‐ATPase activity and decreased the AMPK and SIRT1 expression. Sirtuin 1 overexpression increased Na+/K+‐ATPase activity. 5‐aminoimidazole‐4‐carboxamide‐3‐ribonucleoside (AICAR) upregulated SIRT1 expression and increased Na+/K+‐ATPase activity, which could be partially abolished by splitomicin. Our results suggest that the dysfunction of DCM is related to AGE‐induced Na+/K+‐ATPase activity impairment through a mechanism involving the AMPK/SIRT1 pathway.
    January 28, 2014   doi: 10.1111/1440-1681.12194   open full text
  • Additional Role of Serum 25‐hydroxyvitamin D3 Levels in Atherosclerosis in Chinese Middle‐aged and Elderly Men.
    Yaping Hao, Xiaojing Ma, Yuqi Luo, Jie Ni, Jianxin Dou, Jiaan Zhu, Yuqian Bao, Weiping Jia.
    Clinical and Experimental Pharmacology and Physiology. January 28, 2014
    Perturbed serum vitamin D level has been demonstrated to be associated with increased risk of cardiovascular disease. This study aimed to investigate the association of serum 25‐hydroxyvitamin D3 [25(OH)D3] levels with B ultrasonography‐detected carotid plaque and carotid intima‐media thickness (C‐IMT) in Chinese middle‐aged and elderly men. A total of 1001 men, aged 45‐78 years, were enrolled. Increased C‐IMT was defined as any C‐IMT value in the highest quartile of the study subjects (≥ 0.75 mm). The study population had a median serum 25(OH)D3 level of 14.51 ng/mL (interquartile range: 10.84‐18.67). Subjects with carotid plaque had lower serum 25(OH)D3 levels than those without [13.80 ng/mL (10.82‐17.68) vs. 14.74 ng/mL (10.87–19.08), P = 0.029], and decreasing serum 25(OH)D3 levels were accompanied by increased C‐IMT in both groups [with plaque: 13.24 ng/mL (9.91–16.81) vs. 14.45 ng/mL (11.40–18.51), P < 0.05; without plaque: 13.90 ng/mL (9.99‐17.09) vs. 14.99 ng/mL (11.17‐19.43), P < 0.01]. Multivariate logistic regression analysis showed that serum 25(OH)D3 levels were independently associated with carotid plaque [OR (95% CI): 0.972 (0.946‐0.998), P = 0.032)]. Additionally, serum 25(OH)D3 level was identified as an independent protective factor for increased C‐IMT among subjects with plaque [OR (95% CI): 0.900 (0.849‐0.955), P = 0.001] as well as those without [OR (95% CI): 0.944 (0.908‐0.981), P = 0.004]. These findings collectively suggest that serum 25(OH)D3 levels were inversely associated with atherosclerosis in Chinese middle‐aged and elderly men. This article is protected by copyright. All rights reserved.
    January 28, 2014   doi: 10.1111/1440-1681.12200   open full text
  • GSTA1*‐69C/T and GSTO2*N142D as asthma‐ and allergy‐related risk factors in Italian adult patients.
    Sara Piacentini, Renato Polimanti, Andrea Iorio, Maurizio Cortesi, Fabrizio Papa, Mauro Rongioletti, Giancarlo Maria Liumbruno, Dario Manfellotto, Maria Fuciarelli.
    Clinical and Experimental Pharmacology and Physiology. January 28, 2014
    Asthma and allergies are characterized by variable and subjective symptoms influenced by many genes, molecular mechanisms and environmental factors. The presence of inflammation and oxidative stress in the airways are important biochemical features of asthma and respiratory allergies. Glutathione S‐transferase enzymes (GSTs) play an important role in cellular protection against inflammation and functional genetic polymorphisms in GST genes showed a significant association with asthma and allergy risk. Specifically, our previous study on asthmatic children highlighted GSTA1 and GSTO2 as novel susceptibility loci for asthma. In this study we focused our attention on GSTA1*‐69C/T (rs3957357) and GSTO2*N142D (rs156697) polymorphisms to confirm our previous results in an independent adult study population and to clarify whether GSTA1 and GSTO2 gene polymorphisms are involved in a non‐discriminative pathway towards asthma and respiratory allergy. To accomplish this, we recruited 103 patients with respiratory allergies, 199 patients with asthma, and 200 healthy controls. Genomic DNA extracted from buccal cells was screened for GSTA1*‐69C/T and GSTO2*N142D single nucleotide polymorphisms. The GSTA1*‐69T and GSTO2*D142 variants are both associated with the significant increased risk of asthma, whereas only GSTA1*‐69C/T is significantly associated with allergies. These outcomes confirmed the involvement of GSTO2 loci in asthma, and they suggested that GSTA1 is a common risk factor for asthma and allergies. This article is protected by copyright. All rights reserved.
    January 28, 2014   doi: 10.1111/1440-1681.12201   open full text
  • Increased PTRF/Cavin‐1 expression attenuates PDGFR signaling in senescent human fibroblasts.
    Qian Li, Lin Bai, Ning Liu, Miao Wang, Jun‐Ping Liu, Pingsheng Liu, Yu‐Sheng Cong.
    Clinical and Experimental Pharmacology and Physiology. January 28, 2014
    Our previous study showed that the essential caveolar component PTRF was up‐regulated and promoted caveolae formation in senescent cells. Additionally, we found that overexpression of PTRF increased the number of caveolae and induced cellular senescence. Unresponsiveness to growth factor is one of the fundamental characteristics of senescent cells, although normal levels of receptors and downstream signaling molecules are present in senescent cells. We investigated the role of PTRF in the regulation of growth factor PDGF signaling in young and senescent cells. We first confirmed that PTRF was up‐regulated in senescent human fibroblasts and aged mouse tissues. We then examined the activation of ERK kinases in young and senescent cells after PDGF stimulation. Our results showed that expression of PDGFRs was not altered during cellular senescence. Interestingly, phosphorylation of ERK1/2 was induced upon PDGF stimulation in young, replicating cells but not in senescent cells. Induction of ERK1/2 phosphorylation was impaired in senescent cells and PTRF‐over‐expressed, pre‐senescent cells. Furthermore, our results showed that PTRF interacted with PDGFRs, and this interaction was increased in senescent cells. These results suggest that the unresponsiveness of senescent fibroblasts to PDGF stimulation may be due to increased levels of PTRF and the formation of caveolae, which in turn sequester growth receptors such as the PDGFR and its signaling molecules. This article is protected by copyright. All rights reserved.
    January 28, 2014   doi: 10.1111/1440-1681.12202   open full text
  • Effects Of Acute And Long‐Term Typical Or Atypical Neuroleptics On Morphine‐Induced Behavioural Effects In Mice.
    André W. Hollais, Camilla L. Patti, Karina A. Zanin, Daniela F. Fukushiro, Laís F. Berro, Rita C. Carvalho, Sonia R. Kameda, Roberto Frussa‐Filho.
    Clinical and Experimental Pharmacology and Physiology. January 28, 2014
    It has been suggested that the high prevalence of drug abuse in schizophrenics is related to chronic treatment with typical neuroleptics and dopaminergic supersensitivity that develops as a consequence. Within this context, atypical neuroleptics do not seem to induce this phenomenon. We investigated the effects of acute administration or withdrawal from long‐term administration of haloperidol and/or ziprasidone on morphine‐induced open‐field behaviour in mice. In the first experiment, mice received an injection of haloperidol (1 mg/kg) or several doses of ziprasidone (2, 4 or 6 mg/kg) and had their motor activity quantified in the open‐field. The second experiment aimed to verify the effects of an acute injection of haloperidol (1 mg/Kg) or ziprasidone (6 mg/kg) on 20 mg/kg‐morphine‐induced behaviours in the open‐field. In the third experiment, mice were treated with haloperidol and/or ziprasidone for 20 days. After 72 h, animals received an injection of 20 mg/kg morphine and were exposed to the open‐field. Acute haloperidol or ziprasidone decreased spontaneous open‐field general activity and abolished morphine‐induced locomotor stimulation. Withdrawal from haloperidol or ziprasidone did not modify morphine‐elicited behaviours in the open‐field. It is suggested that withdrawal from neuroleptic treatments does not contribute to morphine acute effects in schizophrenic patients. This article is protected by copyright. All rights reserved.
    January 28, 2014   doi: 10.1111/1440-1681.12203   open full text
  • Inhibition of soluble epoxide hydrolase is renoprotective in 5/6 nephrectomized Ren‐2 transgenic hypertensive rats.
    Petr Kujal, Věra Čertíková Chábová, Petra Škaroupková, Zuzana Husková, Zdena Vernerová, Herbert J Kramer, Agnieszka Walkowska, Elzbieta Kompanowska‐Jezierska, Janusz Sadowski, Kento Kitada, Akira Nishiyama, Sung Hee Hwang, Bruce D. Hammock, John D. Imig, Luděk Červenka.
    Clinical and Experimental Pharmacology and Physiology. January 28, 2014
    The aim of the present study was to test the hypothesis that increasing kidney tissue epoxyeicosatrienoic acids (EETs) concentration by preventing their degradation to the biologically inactive dihydroxyeicosatrienoic acids (DHETEs) using blockade of soluble epoxide hydrolase (sEH), would attenuate the progression of chronic kidney disease (CKD).Ren‐2 transgenic rats (TGR) after 5/6 renal mass reduction (5/6 NX) served as a model of CKD associated with angiotensin II (ANG II)‐dependent hypertension. sEH was inhibited using cis‐4‐[4‐(3‐adamantan‐1‐yl‐ureido)cyclohexyloxy]benzoic acid (c‐AUCB, 3 mg/l of drinking water) for 20 weeks after 5/6 NX. Sham‐operated normotensive transgene‐negative Hannover Sprague‐Dawley (HanSD) rats served as controls. c‐AUCB treatment when applied in TGR subjected to 5/6 NX improved the rats’ survival rate, prevented the increase in blood pressure, retarded the progression of cardiac hypertrophy, reduced proteinuria and the degree of glomerular and tubulointerstitial injury, and reduced the glomerular volume, All these organ‐protective actions were associated with normalization of the intrarenal EETs/DHETEs ratio, an index of availability of biologically active EETs, to levels observed in sham‐operated HanSD rats. There were no significant concurrent alterations of increased intrarenal ANG II content. Taken together, these results show that, first, 5/6 NX TGR exhibit a profound deficiency of intrarenal availability of active epoxygenase metabolites (EETs), which probably contributes to the progression of CKD in this model of ANG II‐dependent hypertension. Second, restoration of intrarenal availability of EETs using long‐term c‐AUCB treatment exhibits substantial renoprotective actions. This article is protected by copyright. All rights reserved.
    January 28, 2014   doi: 10.1111/1440-1681.12204   open full text
  • Resveratrol And Vitamin E Rescued Valproic Acid‐Induced Teratogenicity The Action Mechanism.
    Chiu‐Lan Hsieh, Kuan‐Chou Chen, Ping‐Xiao Lin, Chiung‐Chi Peng, Robert Y. Peng.
    Clinical and Experimental Pharmacology and Physiology. January 28, 2014
    VPA uniquely induced hemorrhagic liposis of the cervical muscles in the chicken embryo model (CEM). Vitamin E (Vit E) and resveratrol (RV) have been cited to exhibit prominent antioxidative and glutathione (GSH)‐protecting effects. We hypothesized that Vit E and RV could ameliorate such kind pathological change. 120 day‐zero fertilized eggs were divided into 10 groups, 12 in each. VPA (60 μM), RV (at 0.2 and 2.0 μM) and Vit E (at 0.2 and 2.0 μM) in several combinations were tested with HH stage‐10 (day 1.5) CEM according to the method as indicated.RV and Vit E at 2.0 μM effectively rescued NTD in early stage CEM and inhibited the malformation rate (8.4 and 5.0%, respectively vs. the control 36.5±3.0%) (p<0.05), suppressing the serum homocysteine and S‐adenosylhomocysteine levels, downregulating the cervical muscular carnitine, triglycerides, H2O2, MDA, IL‐6 and ACC (p<0.05) and upregulating CPT1 (p<0.05) and glutathione (p<0.05). The hemorrhagic liposis of cervical muscles can be alleviated by RV and Vit E. Suggestively, the main action mechanism of RV and Vit E to rescue such VPA‐induced teratogenicity is through suppressing ROS and regenerating GSH. This article is protected by copyright. All rights reserved.
    January 28, 2014   doi: 10.1111/1440-1681.12205   open full text
  • Association of Inflammation, Myocardial Fibrosis, and Cardiac Remodeling in Patients with Mild Aortic Stenosis as Assessed by Biomarkers and Echocardiography.
    Ji Young Park, Sung Kee Ryu, Jae Woong Choi, Kim Min Ho, Jin Hyun Jun, Seung‐Woon Rha, Seong‐Mi Park, Hyo Jeong Kim, Byoung Geol Choi, Yung‐Kyun Noh, Seungwhan Kim.
    Clinical and Experimental Pharmacology and Physiology. January 28, 2014
    The aim of this study is to investigate the relationship of inflammation, myocardial fibrosis, and cardiac remodeling in patients with mild aortic stenosis (AS) as assessed by biomarkers and echocardiography. We consecutively evaluated 32 patients with mild AS and thirty age and gender matched, healthy individuals with normal aortic valves were recruited as control subjects for this study. Baseline echocardiography showed that left ventricular mass index (LVMI, 111.3 ± 26.9 g/m2 vs. 94.5 ± 18.2 g/m2, p‐value = 0.006) and left atrial volume index (LAVI, 27.5 ± 9.0 mm3/mm2 vs. ± 5.2 mm3/mm2, p‐value = 0.005) were significantly higher in patients with mild AS. Left atrial enlargement (LAVI >33 mm3/mm2, 32.4% vs. 3.3%, p‐value = 0.003) and elevated left ventricular filling pressure (E/e’ >15, 50.0% vs. 23.3%, p‐value=0.036) were higher in patients with mild AS. In the patients with mild AS, stepwise, multivariate linear regression analysis showed that the left ventricular end diastolic volume index (LVEDVI) was independently associated with matrix metalloproteinase (MMP)‐1(β= 0.371, p‐value=0.015), aortic valve mean pressure gradient (AVMPG) was independently associated with MMP‐2 (β= 0.19, p‐value=0.019), transforming growth factor (TGF)‐β (β=0.95, p‐value<0.001) and interleukin (IL)‐1(β= 0.17, p‐value=0.019) were independently associated with MMP‐2, and IL‐1 was independently associated with tissue inhibitor of matrix metalloproteinase (TIMP)‐1 (β= 0.68, p‐value=0.001). Myocardial fibrosis in mild AS is independently associated with three factors: LV volume overload, aortic valve pressure overload, and inflammation. This article is protected by copyright. All rights reserved.
    January 28, 2014   doi: 10.1111/1440-1681.12206   open full text
  • Endothelial and neuronal nitric oxide synthases variably modulate the estrogen‐mediated control of blood pressure and cardiovascular autonomic control.
    Mahmoud M. El‐Mas, Abdel A. Abdel‐Rahman.
    Clinical and Experimental Pharmacology and Physiology. January 28, 2014
    We have previously shown that long‐term estrogen (E2) replacement lowers blood pressure (BP) and improves the cardiovascular autonomic control in ovariectomized (OVX) rats. In this study, we investigated whether constitutive and/or inducible nitric oxide synthase (NOS) modulate these E2 effects.We evaluated changes in BP, myocardial contractility index (dP/dtmax), and power spectral indices of hemodynamic variability following selective inhibition of eNOS [N5‐(1‐iminoethyl)‐L‐ornithine; L‐NIO], nNOS (Nω‐propyl‐L‐arginine; NPLA), or iNOS (1400W) in telemetered OVX rats treated for 16 weeks with (OVXE2) or without (control, OVXC) E2.OVXE2 rats exhibited: (i) reduced BP, and increased dP/dtmax, (ii) cardiac parasympathetic dominance as reflected by the reduced low‐frequency (LF, 0.25‐0.75 Hz)/high‐frequency (HF, 0.75‐3 Hz) ratio of interbeat intervals (IBILF/HF), and (iii) reduced LF oscillations of systolic BP, suggesting a reduced vasomotor sympathetic tone.eNOS inhibition (L‐NIO, 20 mg/kg i.p.) elicited a shorter‐lived pressor response in OVXE2, than in OVXC, rats along with reductions in dP/dtmax and increases in the spectral index of spontaneous baroreflex sensitivity (index α). NPLA (1 mg/kg i.p.) reduced BP and increased IBILF/HF ratio in OVXE2, but not OVXC rats. The iNOS inhibitor 1400W (5 mg/kg i.p.) caused no hemodynamic changes in OVXC or OVXE2 rats.Overall, constitutive NOS isoforms exert restraining tonic modulatory BP effects, which encompass eNOS‐mediated reduction and nNOS‐mediated elevation in BP in OVXE2 rats. Baroreflex facilitation, and dP/dtmax reductions might account for the shorter pressor action of L‐NIO in E2‐treated, compared with untreated, OVX rats. This article is protected by copyright. All rights reserved.
    January 28, 2014   doi: 10.1111/1440-1681.12207   open full text
  • Renal vascular structural properties and their alterations by removal of uraemic toxins in a rat model of chronic kidney disease.
    Hidenori Yamazaki, Fumihiro Tomoda, Tsutomu Koike, Hiroyuki Kinuno, Hiroko Sugimori, Hiroshi Inoue, Kenji Bannai, Mikio Sugano, Fuyuhiko Nishijima.
    Clinical and Experimental Pharmacology and Physiology. January 28, 2014
    Renal vascular structural properties and their alterations by removal of uraemic toxins with AST‐120, an oral adsorbent, were examined in subtotal nephrectomised rats. Eight‐ or 9‐week old Sprague‐Dawley rats received 3/4 nephrectomy (n=18) and thereafter were fed 24.5% protein diet with (AST; n=9) or without (No AST; n=9) AST‐120 (0.4 g/100g of body weight). Sham operated rats (Sham; n=9) received the diet without AST. At 21‐22 weeks of age, flow‐pressure and pressure‐glomerular filtration rate relationships (F‐P and P‐GFR) were determined for maximally vasodilated, perfused kidneys. The gradient of F‐P (minimal renal vascular resistance reflecting the overall luminal dimensions of preglomerular and postglomerular vasculature) was lower in No AST than in Sham. By contrast, the X intercept (preglomerular‐to‐postglomerular vascular resistance ratio) and gradient (glomerular filtration capacity) of P‐GFR did not differ between the two groups. The vascular wall and lumen at the interlobular arteries were greater in No AST than in Sham. Although the vascular wall and lumen at the interlobular arteries were less in AST than in No AST, the gradient of F‐P and X intercept of P‐GFR did not differ between the two groups. Contrarily, glomerular filtration capacity was greater in AST than in No AST. In conclusion, the lumen of both preglomerular and postglomerular resistance vessels increased and glomerular filtration capacity failed to increase in subtotal nephrectomised rats. Uraemic toxins could play an important role in the developments of structural alterations in glomeruli rather than renal resistance vessels, in chronic kidney disease. This article is protected by copyright. All rights reserved.
    January 28, 2014   doi: 10.1111/1440-1681.12208   open full text
  • Mathematical modelling of enteric neural motor patterns.
    Jordan D Chambers, Evan A Thomas, Joel C Bornstein.
    Clinical and Experimental Pharmacology and Physiology. January 28, 2014
    The enteric nervous system modulates intestinal behaviours such as motor patterns and secretion. While much is known about different types of neurons and simple reflexes in the intestine, it remains unclear how complex behaviours are generated. Mathematical modelling is an important tool for assisting the understanding of how the neurons and reflexes can be pieced together to generate intestinal behaviours.Models have identified a functional role for slow excitatory post synaptic potentials (EPSPs) by distinguished between fast and slow EPSPs in the ascending excitation reflex. These models also discovered coordinated firing of similarly located neurons as emergent properties of feedforward networks of interneurons in the intestine.A model of the recurrent network of intrinsic sensory neurons identified important control mechanisms to prevent uncontrolled firing due to positive feedback and that the interaction between these control mechanisms and slow EPSPs is necessary for the networks to encode ongoing sensory stimuli. This model also showed that such networks may mediate migrating motor complexes.A network model of VIP neurons in the submucosal plexus found this relatively sparse recurrent network could produce uncontrolled firing under conditions that appear to be related to cholera toxin induced hypersecretion.Abstract modelling of the intestinal fed‐state motor patterns has identified how stationary contractions can arise from a polarised network. These models have also helped predict and explained pharmacological evidence for two rhythm generators and the requirement of feedback from contractions in the circular muscle. This article is protected by copyright. All rights reserved.
    January 28, 2014   doi: 10.1111/1440-1681.12209   open full text
  • Enalapril attenuates ischaemic brain oedema and protects the blood–brain barrier in rats via an anti‐oxidant action.
    Hamdollah Panahpour, Gholam Abbas Dehghani, Shahab Bohlooli.
    Clinical and Experimental Pharmacology and Physiology. January 28, 2014
    In the present study, we investigated the effects of post‐ischaemic angiotensin‐converting enzyme (ACE) inhibition with enalapril on vasogenic oedema formation and blood–brain barrier (BBB) integrity following transient focal cerebral ischaemia in rats.Cerebral ischaemia was induced by 60 min occlusion of the right middle cerebral artery, followed by 24 h reperfusion. Vehicle and non‐hypotensive dose of enalapril (0.03 mg/kg) were administered at the beginning of the reperfusion period. A neurological deficit score (NDS) was determined for all rats at the end of the reperfusion period. Then, brain oedema formation was investigated using the wet–dry weight method and BBB permeability was evaluated on the basis of extravasation of Evans blue (EB) dye. In addition, oxidative stress was assessed by measuring reduced glutathione (GSH) and malondialdehyde (MDA) in brain homogenates. Inhibition of ACE by enalapril significantly reduced NDS and decreased brain oedema formation (P < 0.05 for both). Disruption of the BBB following ischaemia resulted in considerable leakage of EB dye into the brain parenchyma of the ipsilateral hemispheres of vehicle–treated animals. Enalapril significantly (P < 0.05) decreased EB extravasation into the lesioned hemisphere. Enalapril also augmented anti‐oxidant activity in ischaemic brain tissue by increasing GSH concentrations and significantly (P < 0.05) attenuating the increased MDA levels in response to ischaemia. In conclusion, inhibition of ACE with a non‐hypotensive dose of enalapril may protect BBB function and attenuate oedema formation via anti‐oxidant actions. This article is protected by copyright. All rights reserved.
    January 28, 2014   doi: 10.1111/1440-1681.12210   open full text
  • Aerobic interval training protects against myocardial infarction‐induced oxidative injury by enhancing antioxidase system and mitochondrial biosynthesis.
    Hong‐Ke Jiang, Yi Miao, You‐Hua Wang, Mei Zhao, Zhi‐Hui Feng, Xiao‐Jiang Yu, Jian‐Kang Liu, Wei‐Jin Zang.
    Clinical and Experimental Pharmacology and Physiology. January 28, 2014
    Aerobic interval training (AIT) exerts beneficial effects on cardiovascular disease. However, its cardioprotective mechanisms are not fully understood. This study was designed to evaluate AIT‐mediated antioxidation by focusing on antioxidase and mitochondrial biogenesis in rats post‐myocardial infarction (MI). Sprague‐Dawley rats were divided into three groups: sham‐operated (CON), MI and MI+AIT. Myocardial microstructure and function, oxidative stress markers, mitochondrial antioxidase, phase II enzymes and mitochondrial biogenesis were assessed. The nuclear factor‐erythroid 2‐related factor (Nrf) and phosphorylated 5’‐AMP‐activated kinase (AMPK) levels were determined. The antioxidative gene sirtuin (SIRT) 3 and pro‐survival phosphatidylinositol‐3 kinase (PI3K)‐protein kinases B (Akt) signaling cascades were also evaluated. Compared with CON, post‐MI rats demonstrated noticeable microstructure injury, cardiac dysfunction and oxidative damage. In addition, decreased mitochondrial antioxidase contents, phase II enzyme (except heme oxygenase‐1) expressions and mitochondrial biogenesis were observed as well as reduced protein levels of the regulators Nrf2 and phosphorylated AMPK. SIRT3 levels and PI3K‐Akt signaling were also suppressed. These detrimental modifications were considerably ameliorated by AIT, as evidenced by increased antioxidase and elevated mitochondrial biogenesis. Nrf2 and AMPK phosphorylation were also increased after AIT, accompanied by SIRT3 upregulation and PI3K‐Akt signaling activation. Moreover, PI3K inhibitor treatment partly attenuated AIT‐elicited increased Nrf2 levels and AMPK phosphorylation.AIT can effectively alleviate MI‐induced oxidative injury, which may closely correlate with antioxidase system activation and mitochondrial biosynthesis. Increased SIRT3 expression and PI3K‐Akt signaling activation may play key roles in AIT‐mediated antioxidation. These results open up new avenues for exercise intervention therapies for MI patients. This article is protected by copyright. All rights reserved.
    January 28, 2014   doi: 10.1111/1440-1681.12211   open full text
  • Differential trafficking of saccharidic probes following aspirin in clinical tests of intestinal permeability in young healthy women.
    Ivana R Sequeira, Roger G Lentle, Marlena C Kruger, Roger D Hurst.
    Clinical and Experimental Pharmacology and Physiology. January 28, 2014
    The effects of inflammatory changes on the absorption of different‐sized probes and their permeability ratios are poorly understood. The aim of the present study was to determine the effects of a pharmacological agent on the permeability of the gut mucosa to saccharidic probes of larger and smaller molecular weight. Permeability was assessed by half‐hourly urinary excretion of a combined dose of d‐mannitol, l‐rhamnose and lactulose following consumption of a single 600 mg dose of aspirin and compared with a placebo in a cross‐over study in 20 healthy female volunteers. The temporal patterns of excretion of all probes were bimodal, being best fitted by polynomial functions. The relatively small early peak was evident for at least 4 h for smaller sugars, but was less evident with lactulose, being overshadowed by a larger second peak. These conclusions were further supported by separate analyses of the segments of the temporal plots between 2.5 and 4 h and between 4.5 and 6 h. The forms of these curves did not change significantly following dosing with aspirin. A greater proportion of the total dose of mannitol than rhamnose was excreted over the collection period. Following the consumption of aspirin, the cumulative rate of excretion of the smaller sugars (i.e. mannitol and rhamnose) was significantly reduced whereas that of lactulose was increased over the 6 h collection period. Aspirin has opposite effects on the absorption of larger and smaller probes, influencing the outcome of the test. These results have important consequences for the design and comparison of clinical tests of permeability.
    January 28, 2014   doi: 10.1111/1440-1681.12163   open full text
  • Neuroprotective effects of ebselen in traumatic brain injury model: involvement of nitric oxide and p38 mitogen‐activated protein kinase signalling pathway.
    Liang Wei, Yanfei Zhang, Cheng Yang, Qi Wang, Zhongwei Zhuang, Zhiyang Sun.
    Clinical and Experimental Pharmacology and Physiology. January 28, 2014
    Previous investigations have found that ebselen is able to treat neurodegenerative diseases caused by radical and acute total cerebral ischaemia. The aim of the present study was to investigate the neuroprotective effects of ebselen in a traumatic brain injury (TBI) model. Ninety Sprague‐Dawley rats were randomly divided into five groups (n = 18 in each): (i) sham operation; (ii) an injury model group; (iii) low‐dose (3 mg/kg) ebselen‐treated group; (iv) a moderate‐dose (10 mg/kg) ebselen‐treated group; and (v) a high‐dose (30 mg/kg) ebselen‐treated group. The TBI model was created according using a modified weight‐drop model. Neurological severity score (NSS), brain water content and histopathological deficits were assessed as parameters of injury severity. Expression of nitric oxide (NO), inducible NO synthase (iNOS) mRNA, Toll‐like receptor (TLR) and phosphorylated (p‐) p38 mitogen‐activated protein kinase (MAPK) were examined by chemical colorimetry, quantitative polymerase chain reaction and western blotting 24 h after intragastric ebselen administration. Rats in the TBI model group exhibited markedly more severe neurological injury (higher NSS, more brain water content and more histopathological deficits) than those in the sham‐operated group. Ebselen treatment significantly ameliorated the neurological injury of TBI rats in a dose‐dependent manner. Moreover, ebselen significantly reduced the NO and iNOS mRNA levels and inhibited TLR4 and p‐p38 MAPK expression, indicating the involvement of NO and p38 MAPK signalling pathways in the neuroprotection afforded by ebselen. In conclusion, ebselen ameliorated neurological injury, possibly by reducing NO levels and modulating the TLR4‐mediated p38 MAPK signalling pathway. Therefore, ebselen may have potential to treat secondary injuries of TBI.
    January 28, 2014   doi: 10.1111/1440-1681.12186   open full text
  • Current approach to masked hypertension: From diagnosis to clinical management.
    Eamon Dolan, Kirstyn James.
    Clinical and Experimental Pharmacology and Physiology. November 28, 2013
    The term masked hypertension phenomenon was first described by the late Professor Thomas Pickering and is commonly defined as having a normal clinic blood pressure (BP) but an elevated “out of office” reading. In the main these elevated readings have been provided through ambulatory blood pressure monitoring (ABPM) but sometimes home BP monitoring is used. It is now largely accepted that ABPM gives a better classification of risk than clinic BP. Thus the elevated ABPM levels should relate to higher cardiovascular risk and it follows that these people might be regarded as being genuinely hypertensive and at higher cardiovascular risk. The problem for clinical practice is how to identify and manage these subjects. The phenomenon should be suspected in subjects who have had an elevated clinic BP at some time, in young subjects with normal or normal‐high clinic BP who have early left ventricular hypertrophy, in subjects with a family history of hypertension in both parents, patients with multiple risks for cardiovascular disease and perhaps diabetic patients. It appears to be more prevalent in subjects of male gender, with younger age, higher heart rate, obesity or high cholesterol levels and in smokers. Those with masked hypertension are at higher risk of events such as stroke and have a higher prevalence of target organ damage, for example, nephropathy. In conclusion most of the debate around this topic relates to its reliable identification. Given the higher ambulatory readings there is an increases cardiovascular risk making this diagnosis important. This article is protected by copyright. All rights reserved.
    November 28, 2013   doi: 10.1111/1440-1681.12190   open full text
  • Pulse pressure variation does not reflect stroke volume variation in mechanically ventilated rats with lipopolysaccharide‐induced pneumonia.
    Thomas GV Cherpanath, Lonneke Smeding, Wim K Lagrand, Alexander Hirsch, Marcus J Schultz, AB Johan Groeneveld.
    Clinical and Experimental Pharmacology and Physiology. October 28, 2013
    1.The present study examined the relationship between centrally measured stroke volume variation (SVV) and peripherally derived pulse pressure variation (PPV) in the setting of increased total arterial compliance (CArt). 2.Ten male Wistar rats were anaesthetised, paralysed and mechanically ventilated before being randomized to receive intrapulmonary lipopolysaccharide (LPS) or no LPS. Pulse pressure (PP) was derived from the left carotid artery, whereas stroke volume (SV) was measured directly in the left ventricle. Values of SVV and PPV were calculated over three breaths. Balloon inflation of a catheter positioned in the inferior vena cava was used, for a maximum of 30 s, to decrease preload while the SVV and PPV measurements were repeated. Values of CArt were calculated as SV/PP. 3.Intrapulmonary LPS increased CArt and SV. Values of SVV and PPV increased in both LPS‐treated and untreated rats during balloon inflation. There was a correlation between SVV and PPV in untreated rats before (r = 0.55; P = 0.005) and during (r = 0.69; P < 0.001) occlusion of the vena cava. There was no such correlation in LPS‐treated rats either before (r = –0.08; P = 0.70) or during (r = 0.36; P = 0.08) vena cava occlusion. 4.In conclusion, under normovolaemic and hypovolaemic conditions, PPV does not reflect SVV during an increase in CArt following LPS‐induced pneumonia in mechanically ventilated rats. Our data caution against their interchangeability in human sepsis. This article is protected by copyright. All rights reserved.
    October 28, 2013   doi: 10.1111/1440-1681.12187   open full text
  • The renin–angiotensin system from conception to old age: the good, the bad and the ugly.
    Eugenie R Lumbers, Kirsty G Pringle, Yu Wang, Karen J Gibson.
    Clinical and Experimental Pharmacology and Physiology. October 27, 2013
     The renin–angiotensin system (RAS) plays a critical role in placentation and nephrogenesis. Failure to thrive during intrauterine life, possibly related to placental dysfunction and impaired expression of the renal RAS, as well as prematurity, results in smaller kidneys at birth and reduced nephron number. The remaining nephrons are therefore hyperfiltering from birth. Hyperfiltration, infections and Type 2 diabetes cause glomerular and tubular fibrosis, leading to further reductions in nephron number.  The intrarenal RAS plays a key role in promoting tubulointerstitial fibrosis. Low birth weight and a high incidence of preterm birth program Indigenous children for early onset renal disease in adult life. Indigenous Australians have 404 000 fewer nephrons than non‐Indigenous Australians. This, coupled with the high incidence of infectious diseases (particularly acute post‐streptococcal glomerulonephritis) and the increasing prevalence of Type 2 diabetes, explains why end‐stage renal disease is of epidemic proportions in Indigenous Australians.  The existence of RAS gene polymorphisms and inflammatory cytokines may further potentiate susceptibility to renal disease in Indigenous Australians.
    October 27, 2013   doi: 10.1111/1440-1681.12098   open full text
  • Introduction: Celebrating Emeritus Scientia Professor Eugenie R Lumbers AM and Professor Caroline McMillen.
    Janna L. Morrison, Eugenie Lumbers, Laura Bennet, Jane Black.
    Clinical and Experimental Pharmacology and Physiology. October 27, 2013
    There is no abstract available for this paper.
    October 27, 2013   doi: 10.1111/1440-1681.12180   open full text
  • Reciprocal relationship between plasma ghrelin level and arterial stiffness in hypertensive subjects.
    Yin‐Tao Zhao, Hai‐Bo Yang, Ling Li, Ke Gao, Peng‐Fei Li, Wei‐Wei Xie.
    Clinical and Experimental Pharmacology and Physiology. October 27, 2013
    Arterial stiffness, considered an independent predictor of cardiovascular morbidity and mortality, is closely associated with hypertension. Futhermore, the role of ghrelin in the development of hypertension has been widely recognized. The purpose of the present study was to explore the potential relationship between circulating ghrelin and arterial stiffness in hypertensive subjects. A total of 192 patients with primary hypertension and 107 normotensive (NT) control subjects were enrolled in the present cross‐sectional study. Plasma ghrelin was determined by ELISA. Arterial stiffness was assessed by brachial–ankle pulse wave velocity (baPWV) and the augmentation index (AIx). Both baPWV and AIx values were markedly higher in the hypertensive compared with NT group (P < 0.01). In contrast, plasma ghrelin concentrations were significantly lower in hypertensive patients compared with NT subjects (P < 0.01). Plasma ghrelin concentrations were negatively correlated with age (odds ratio (OR) −1.836; P < 0.001), smoking (OR −1.347; P = 0.042), baPWV (OR −1.762; P < 0.001) and AIx (OR −1.516; P = 0.005), but positively associated with fasting plasma glucose (OR 1.293; P = 0.047) and HbA1c (OR 1.413; P = 0.025). The inverse correlation between circulating ghrelin and the extent of arterial stiffness suggests that ghrelin is an independent determinant of arterial stiffness, even after adjustment for confounding cardiovascular risk factors, and it actively participates in the pathophysiology of arterial stiffness in hypertensive subjects.
    October 27, 2013   doi: 10.1111/1440-1681.12165   open full text
  • Fructose, pregnancy and later life impacts.
    Timothy RH Regnault, Sheridan Gentili, Ousseynou Sarr, Carla R Toop, Deborah M Sloboda.
    Clinical and Experimental Pharmacology and Physiology. October 27, 2013
    Fructose is an increasingly common constituent of the Westernized diet due to cost and production efficiencies. Although an integral component of our pre‐industrial revolution diet, over the past two decades human and animal studies have highlighted that excessive fructose intake appears to be associated with adverse metabolic effects. Excessive intake of fructose is the combined result of increased total energy consumption and increased portion sizes of foods, which often incorporate the fructose‐containing sugars sucrose and high‐fructose corn‐syrup (HFCS). The adverse metabolic effects following excessive fructose consumption have become a hot topic in mainstream media and there is now rigorous scientific debate regarding periods of exposure, dosage levels, interactive effects with other sugars and fats and mechanisms underlying the actions of fructose. There is still a degree of controversy regarding the extent to which sugars such as sucrose and HFCS have contributed to the current epidemic of obesity and diabetes. Furthermore, an increasing number of infants are being exposed to sugar‐sweetened food and beverages before birth and during early postnatal life, highlighting the importance of determining the long‐term effects of this perinatal exposure on the developing offspring. There are limited human observational and controlled studies identifying associations of excessive sweetened food and beverage consumption with poor pregnancy outcomes. Animal research has demonstrated an increased incidence of gestational diabetes as well as altered maternal, fetal and offspring metabolic function, although the long‐term effects and the mechanism underlying these perturbations are ill defined. This review aims to understand the role of early life fructose exposure in modifying postnatal risk of disease in the offspring, focusing on fructose intake during pregnancy and in early postnatal life.
    October 27, 2013   doi: 10.1111/1440-1681.12162   open full text
  • Haploinsufficiency of endogenous parathyroid hormone‐related peptide impairs bone fracture healing.
    Yin‐He Wang, Yong Qiu, Xiao‐Dong Han, Jin Xiong, Yi‐Xin Chen, Hong‐Fei Shi, Andrew Karaplis.
    Clinical and Experimental Pharmacology and Physiology. October 27, 2013
    Previous studies have demonstrated that endogenous parathyroid hormone‐related peptide (PTHrP) plays a central role in the physiological regulation of bone formation. However, it is unclear whether endogenous PTHrP plays an important function in enhancing bone fracture healing. To determine whether endogenous PTHrP haploinsufficiency impaired bone fracture healing, closed mid‐diaphyseal femur fractures were created in 8‐week‐old wild‐type and Pthrp+/− mice. Callus tissue properties were analysed 1, 2 and 4 weeks after fracture by radiography, histology, histochemistry, immunohistochemistry and molecular biology. The size of the calluses was reduced 2 weeks after fracture, and the fracture repairs were poor 4 weeks after fractures, in Pthrp+/− compared with wild‐type mice. Cartilaginous callus areas were reduced 1 week after fracture, but were increased 2 weeks after fracture in Pthrp+/− mice. There was a reduction in the number of ostoblasts, alkaline phosphatase (ALP)‐positive areas, Type I collagen immunopositive areas, mRNA levels of ALP, Runt‐related transcription factor 2 (Runx2) and Type I collagen, Runx2 and insulin‐like growth factor‐1 protein levels, the number of osteoclasts and the surface in callus tissues in Pthrp+/− compared with wild‐type mice. These results demonstrate that endogenous PTHrP haploinsufficiency impairs the fracture repair process by reducing cartilaginous and bony callus formation, with downregulation of osteoblastic gene and protein expression and a reduction in endochondral bone formation, osteoblastic bone formation and osteoclastic bone resorption. Together, the results indicate that endogenous PTHrP plays an important role in fracture healing.
    October 27, 2013   doi: 10.1111/1440-1681.12161   open full text
  • Kidney regeneration by non‐platelet RNA‐containing particle‐derived cells.
    Wuyi Kong, Mu Nuo, Xiao Ping Zhu, Xiu Juan Han, Xian Wang.
    Clinical and Experimental Pharmacology and Physiology. October 27, 2013
    We found a group of non‐platelet RNA‐containing particles (NPRCPs) in human umbilical cord blood. These particles can aggregate, fuse and become non‐nucleated cells when cocultured with nucleated cells in vitro. The non‐nucleated cells further differentiate into nucleated cells expressing octamer binding transcription factor 4 (OCT4). The NPRCPs are approximately 1–5 μm in diameter, have a thin bilayer membrane, contain short RNAs and microRNAs and express OCT4, sex‐determining region Y 2 (SOX2) and DEAD box polypeptide 4 (DDX4). To confirm the function of NPRCPs in vivo, we examined the effects of tail vein‐injected green fluorescent protein (GFP)‐labelled NPRCPs on mouse kidneys damaged by prior ischaemia and reperfusion from Day 1 to Week 6. Within 1 day of injection of NPRCPs, immunofluorescence and immunohistochemistry revealed a large number of extravasated NPRCPs in the renal calyces, damaged glomeruli and duct tubules. During the course of regeneration, NPRCPs fused into large, non‐nucleated cellular structures that further became large nucleated cells to regenerate multicellular kidney tubules. In addition, many NPRCPs became tiny nucleated cellular structures that further differentiated into interstitial cells in connective tissue. The extravasated NPRCPs also arranged themselves into non‐cell glomerular structures before further regenerating into nucleated cells of the glomerulus. In conclusion, the results demonstrate that, via different patterns of differentiation, NPRCP‐derived cells can regenerate mouse kidney tissue damaged by ischaemia.
    October 27, 2013   doi: 10.1111/1440-1681.12164   open full text
  • Regulation of fetal lung development in response to maternal overnutrition.
    Mitchell Lock, Erin V McGillick, Sandra Orgeig, I Caroline McMillen, Janna L Morrison.
    Clinical and Experimental Pharmacology and Physiology. October 27, 2013
    With the worldwide obesity epidemic, the proportion of women entering pregnancy overweight or obese has increased significantly in recent years. Babies born to obese women are at an increased risk of respiratory complications at birth and in childhood. In addition to maternal diabetes, there are a number of metabolic changes that the fetus of an overnourished mother experiences in utero that may modulate lung development and represent the mechanisms underlying the increased risk of respiratory complications. Herein we highlight a series of factors associated with the intrauterine environment of an overnourished mother that may impact on fetal lung development and lead to an increased risk of complications at birth or in postnatal life.
    October 27, 2013   doi: 10.1111/1440-1681.12166   open full text
  • Excess nutrient supply in early life and its later metabolic consequences.
    Shalini Ojha, Vivek Saroha, Michael E. Symonds, Helen Budge.
    Clinical and Experimental Pharmacology and Physiology. October 27, 2013
    Suboptimal nutrition in early life, both in utero and during infancy, is linked to increased risk of adult obesity and its associated adverse metabolic health problems. Excess nutrient supply during early life can lead to metabolic programming in the offspring. Such overnutrition can occur in the offspring of obese mothers, the offspring of mothers who gain excess weight during gestation, infants of diabetic mothers and infants who undergo rapid growth, particularly weight gain, during early infancy. Postnatal overnutrition is particularly detrimental for infants who are born small for gestational age, who are overfed to attain ‘catch‐up growth’. Potential mechanisms underlying metabolic programming that results from excess nutrition during early life include resetting of hypothalamic energy sensing and appetite regulation, altered adipose tissue insulin sensitivity and impaired brown adipose tissue function. More detailed understanding of the mechanisms involved in metabolic programming could enable the development of therapeutic strategies for ameliorating its ill effects. Research in this field could potentially identify optimal and appropriate preventative interventions for a burgeoning population at risk of increased mortality and morbidity from obesity and its concomitant metabolic conditions.
    October 27, 2013   doi: 10.1111/1440-1681.12061   open full text
  • Impact of preterm birth and bronchopulmonary dysplasia on the developing lung: Long‐term consequences for respiratory health.
    Megan O'Reilly, Foula Sozo, Richard Harding.
    Clinical and Experimental Pharmacology and Physiology. October 27, 2013
    Preterm birth affects 8–10% of human pregnancies and is a major cause of long‐term disability. Individuals who are born very preterm, especially if they develop bronchopulmonary dysplasia (BPD), have an increased risk of impaired lung function in infancy, childhood and adulthood, as well as an increased risk of respiratory illness. Our aim is to briefly review current understanding of the basis for long‐term impairments in lung function and respiratory health following preterm birth and BPD. Histopathology of the lungs of infants and children following preterm birth and BPD shows altered development of the lung parenchyma, conducting airways and pulmonary vasculature. Owing to improvements in the care of preterm infants, especially the use of exogenous surfactant and lower concentrations of administered oxygen, lung pathology following preterm birth and BPD is less severe than in the past. Recent studies indicate that very preterm birth and BPD can lead to hyperplasia of airway smooth muscle, impaired alveolarization, pulmonary inflammation and an increase in pulmonary artery muscularization. Imaging of adult lungs suggests that the deficit in alveoli can persist into later life. Long‐term lung injury apparently relates to the use of mechanical ventilation and the use of supplemental oxygen in infancy. Impaired lung function in later life is due to airway hyper‐reactivity and fewer alveoli, resulting in reductions in the surface area for gas exchange and physical support for bronchioles. Because the incidence of preterm birth is not declining, it will continue to be a major cause of respiratory ill‐health in adults.
    October 27, 2013   doi: 10.1111/1440-1681.12068   open full text
  • Developmental programming: Variations in early growth and adult disease.
    Linda A Gallo, Melanie Tran, Karen M Moritz, Mary E Wlodek.
    Clinical and Experimental Pharmacology and Physiology. October 27, 2013
     Suboptimal conditions in utero are associated with the development of adult‐onset diseases in offspring. Uteroplacental insufficiency in rats is a well‐established animal model used to mimic and study the effects of developmental insults relevant to countries of abundant nutrient supply. However, wide‐ranging outcomes for the offspring are apparent between the different investigators that use this model and also between cohorts generated in our laboratory.  We aimed to explore the reasons for variability in rat models of uteroplacental insufficiency between different investigators and also between our own animal cohorts. We suggest differences in growth and disease development reflect uniqueness in susceptibility and highlight the complexity of interactions between genetic potential and environmental exposures.  The impact of adverse exposures in utero has been described as having far‐reaching effects that extend well beyond the first, directly exposed generation. However, the resulting phenotypes are not consistent between generations. This suggests that programmed effects are established de novo in each generation and challenges the prediction of disease.  Characterization of growth and disease in the numerous rat models has led to our understanding of the impact of early life experiences on adult health. In order to drive the development of preventative and/or treatment strategies, future studies should focus on identifying the initial cause(s) of uteroplacental insufficiency, including genetic origins and the influence of poor diets.
    October 27, 2013   doi: 10.1111/1440-1681.12092   open full text
  • Chronic intrauterine exposure to endotoxin does not alter fetal nephron number or glomerular size.
    Danica Ryan, Anzari Atik, Robert Matteo, Richard Harding, Mary J Black.
    Clinical and Experimental Pharmacology and Physiology. October 27, 2013
     A reduced nephron endowment early in life adversely impacts on long‐term functional reserve in the kidney. A recent study has shown that acute exposure to chorioamnionitis during late gestation can adversely impact on nephrogenesis. The present study aimed to examine the effects of chronic, low‐dose endotoxin exposure in utero, during the period of nephrogenesis, on nephron number and glomerular size in preterm lambs.  Ewes were administered either endotoxin (lipopolysaccharide; 1 mg/day) or saline at 110–133 days of gestation (term approximately 147 days) via surgically implanted osmotic minipumps within the amniotic cavity. The ewes were induced to deliver preterm at 133 days gestation and the kidneys of the lambs were analysed at 8 weeks after term‐equivalent age. Nephron number per kidney was determined using a combined optical disector and fractionator stereological approach; renal corpuscle size was also measured stereologically.  At 8 weeks after term‐equivalent age there was no significant effect of in utero exposure to endotoxin on bodyweight or kidney weight and there were no significant differences in nephron number, nephron density or renal corpuscle volume between groups.  We conclude that chronic intrauterine inflammation during the period of nephrogenesis may not adversely impact on the number of nephrons formed within the kidney or on the volume of the renal corpuscle.
    October 27, 2013   doi: 10.1111/1440-1681.12095   open full text
  • Estimating glomerular number: Why we do it and how.
    John F Bertram.
    Clinical and Experimental Pharmacology and Physiology. October 27, 2013
     There is currently much interest in determining the number of glomeruli, and thereby nephrons, in the kidney.  Researchers have been trying to count glomeruli since the 19th century and currently four general approaches are available: (i) acid maceration; (ii) counting glomerular profiles in histological sections; (iii) model‐based stereology; and (iv) design‐based stereology.  Although design‐based stereological methods are generally considered the gold‐standard method, all current methods have limitations. A new approach using magnetic resonance imaging has recently been described and may ultimately enable glomerular imaging and quantification in vivo.  This report considers the advantages and disadvantages of current methods for counting glomeruli and describes the new magnetic resonance approach. In addition, a method for counting glomeruli in developing kidneys is described.
    October 27, 2013   doi: 10.1111/1440-1681.12133   open full text
  • Neural plasticity and the Kennard principle: does it work for the preterm brain?
    Laura Bennet, Lotte Van Den Heuij, Justin M Dean, Paul Drury, Guido Wassink, Alistair Jan Gunn.
    Clinical and Experimental Pharmacology and Physiology. October 27, 2013
     The Kennard principle suggests that the immature brain should be more able to recover from injury than the more developed brain. Curiously, preterm infants continue to have a high rate of debilitating neurodevelopmental handicaps despite a progressive improvement in structural damage to the brain, from acute necrotic injury of the periventricular white matter, with axonal loss in historical cohorts, to diffuse gliosis with trivial axonal damage.  In the present review we examine recent evidence that disability after preterm birth is largely mediated by disturbed development of neuronal connections. Potential mechanisms include impaired white matter maturation associated with gliosis, suboptimal neuronal maturation, adverse effects of infection/inflammation on the cell environment, exposure to clinical therapies that modulate brain function (including maternal glucocorticoids), upregulation of physiological apoptosis and loss or misprogramming of progenitor cells in the subventricular zone.  These findings suggest that insults during this critical phase alter the trajectory of brain development and that a key focus of basic science and clinical research should be to understand neuronal connectivity, as well as the triggers of cell death.
    October 27, 2013   doi: 10.1111/1440-1681.12135   open full text
  • Endothelial caveolar subcellular domain regulation of endothelial nitric oxide synthase.
    Jayanth Ramadoss, Mayra B Pastore, Ronald R Magness.
    Clinical and Experimental Pharmacology and Physiology. October 27, 2013
    Complex regulatory processes alter the activity of endothelial nitric oxide synthase (eNOS) leading to nitric oxide (NO) production by endothelial cells under various physiological states. These complex processes require specific subcellular eNOS partitioning between plasma membrane caveolar domains and non‐caveolar compartments. Translocation of eNOS from the plasma membrane to intracellular compartments is important for eNOS activation and subsequent NO biosynthesis. We present data reviewing and interpreting information regarding: (i) the coupling of endothelial plasma membrane receptor systems in the caveolar structure relative to eNOS trafficking; (ii) how eNOS trafficking relates to specific protein–protein interactions for inactivation and activation of eNOS; and (iii) how these complex mechanisms confer specific subcellular location relative to eNOS multisite phosphorylation and signalling. Dysfunction in the regulation of eNOS activation may contribute to several disease states, in particular gestational endothelial abnormalities (pre‐eclampsia, gestational diabetes etc.), that have life‐long deleterious health consequences that predispose the offspring to develop hypertensive disease, Type 2 diabetes and adiposity.
    October 27, 2013   doi: 10.1111/1440-1681.12136   open full text
  • Roles played by histamine in strenuous or prolonged masseter muscle activity in mice.
    Hiroyuki Yoneda, Fukie Niijima‐Yaoita, Masahiro Tsuchiya, Hiroyuki Kumamoto, Makoto Watanbe, Hiroshi Ohtsu, Kazuhiko Yanai, Takeshi Tadano, Keiichi Sasaki, Shunji Sugawara, Yasuo Endo.
    Clinical and Experimental Pharmacology and Physiology. October 18, 2013
    1. Bruxism and/or clenching, resulting in fatigue or dysfunction of masseter muscles (MM), may cause temporomandibular disorders. Functional support of the microcirculation is critical for prolonged muscle activity. Histamine is a regulator of the microcirculation and is supplied by release from its stores and/or by de novo production via the induction of histidine decarboxylase (HDC). IL‐1, a cytokine involved in temporomandibular disorders, is an inducer of HDC. We examined the roles of histamine, HDC, and IL‐1 in MM activity. 2. Experiments were conducted using our R+G+ model. A mouse restrained (R+) inside a narrow cylinder (front end blocked with a thin plastic strip) gnaws away (G+) the strip to escape, and the strip's weight‐reduction serves as an index of MM activity. 3. Fexofenadine (peripherally acting histamine H1‐receptor antagonist) reduced MM activity in normal mice. H1‐receptor‐deficient mice and HDC‐deficient mice displayed low MM activity. Prolonged R+G+ induced HDC activity in MM. Mast cell‐deficient mice exhibited strikingly low HDC induction in MM (and also in quadriceps femoris muscle) in response to muscle activity or IL‐1β. Mast cells were present around blood vessels and nerves in the epimysium and perimysium of MM. 4. These results, together with others reported previously, suggest that (i) peripheral histamine supports strenuous MM activity, (ii) strenuous MM activity stimulates mast cells to release histamine and to induce HDC (which replenishes their histamine pool, possibly through mediation by IL‐1), and (iii) peripheral histamine H1‐receptor antagonists may be effective at treating temporomandibular disorders or preventing prolonged clenching and/or bruxism. This article is protected by copyright. All rights reserved.
    October 18, 2013   doi: 10.1111/1440-1681.12167   open full text
  • Nicorandil improves electrical remodeling, leading to the prevention ofelectrically induced ventricular tachyarrhythmia in a mouse model of desmin‐related cardiomyopathy.
    Naoko Matsushita, Masamichi Hirose, Atsushi Sanbe, Yukiko Kondo, Yasuyuki Irie, Eiichi Taira.
    Clinical and Experimental Pharmacology and Physiology. October 10, 2013
    Transgenic (R120G TG) mice with overexpression of an arg120gly missense mutation in HSPB5 display desmin‐related cardiomyopathy. Recently, the cardioprotective effect of nicorandil has been shown to prolong survival in R120G TG mice. However, whether the TG mice exhibit ventricular arrhythmias and nicorandil can inhibit these arrhythmias remains unknown. We examined the effects of chronic administration of nicorandil on ventricular electrical remodeling and arrhythmias in R120G TG mice. Nicorandil (15mg/kg/day) or vehicle was orally administered in R120G TG mice from 5 weeks to 30 weeks of age. Electrocardiogram (ECG) and optical action potentials were recorded from R120G TG mouse hearts. We examined the expression of ventricular connexin43 and cardiac Na+ channel (Nav1.5) in the TG mice. All ECG parameters tested were prolonged in R120G TG mice. Nicorandil improved the prolonged P, PQ, and QRS complex intervals in R120G TG mice. Interestingly, impulse conduction slowing and increases in the expression of total and phosphorylatedconnexin43 and Nav1.5 were observed in R120G TG ventricles. Nicorandil improved ventricular impulse conduction slowing and normalized the increased protein expression levels of total and phosphorylated connexin43,but not of Nav1.5,in R120G TG mouse hearts. The electrical rapid pacing at the ventricle induced ventricular tachyarrhythmias (VT) in 6 of 8 R120G TG mouse hearts but in none of 8 nicorandil‐treated R120G TG mouse hearts (p<0.05). These findings demonstrated that nicorandil inhibitedcardiac electrical remodeling and theprevention of VT by nicorandil was associated with the normalization of connexin43 expression in this model. This article is protected by copyright. All rights reserved.
    October 10, 2013   doi: 10.1111/1440-1681.12185   open full text
  • Probucol Attenuates Ethanol‐induced Liver Fibrosis in Rats through Inhibition of Oxidative Stress, Extracellular Matrix Protein Accumulation and Cytokine Production.
    Xuesong Su, Yanqiu Wang, Guangyu Zhou, Xu Yang, Rui Yu, Yan Lin, Changqing Zheng.
    Clinical and Experimental Pharmacology and Physiology. October 03, 2013
    1.Liver fibrosis is characterized by excessive accumulation of extracellular matrix (ECM) proteins in liver. Probucol, a lipid lowering drug, was found to prevent liver injury in rats treated with carbon tetrachloride (CCl4). In the present study, we investigated whether probucol exerts protection against liver fibrosis in rats treated with ethanol and CCl4. 2.Thirty rats were randomly divided into five groups. Group I and II were served as the normal control and the model of liver fibrosis. Groups III‐V were treated with probucol at a dose of 250, 500, 1000 mg/kg body weight, respectively. Rats in Group II were fed a complex diet that includes alcohol, corn oil and pyrazole, and intraperitoneally injected with CCl4 to induce hepatic fibrosis. Blood was obtained to assess markers of liver function. Liver samples were collected to evaluate mRNA and protein expression, histological alteration and oxidative stress. 3.Probucol significantly attenuated the histological alterations and improved liver function in the model group. The expression levels of α‐SMA and collagen I were decreased in probucol treatment groups. Moreover, the increases of oxidative stress, ECM protein accumulation and cytokine production in hepatic fibrosis were dramatically suppressed by probucol. Finally, probucol inhibited the activation of the ERK signaling pathway in liver fibrosis. 4.Our findings revealed that probucol attenuated ethanol‐induced liver fibrosis through inhibition of oxidative stress, ECM protein accumulation and cytokine production. These data suggest that probucol may be useful for the prevention and treatment of hepatic fibrosis. This article is protected by copyright. All rights reserved.
    October 03, 2013   doi: 10.1111/1440-1681.12182   open full text
  • Hemodynamic Consequences of Recurrent Insulin ‐ Induced Hypoglycemia.
    Syed Quadri, Priyanka Prathipati, Debra W. Jackson, Keith E. Jackson.
    Clinical and Experimental Pharmacology and Physiology. October 03, 2013
    1.Recurring insulin‐induced hypoglycemia (RIIH) often occurs during the therapeutic management of insulin dependent diabetes mellitus. Controversy currently exists in the literature as to the ability of insulin and/or hypoglycemia to promote hypertension. Could insulin and/or hypoglycemia promote adverse pressor effects; if so, under what conditions and through what mechanism? Thus, the current study was performed to evaluate the hypothesis that recurrent insulin‐induced hypoglycemia produces hypertension via an induction in HO‐1 promoting a significant increase in endogenous CO. 2.Male Sprague Dawley rats were treated for 2 weeks with varying doses of subcutaneous insulin injections (1, 3, 5, 7, and 9 U/kg body weight) or vehicle and fed normal rat chow or zinc diet (1mM) for 2 weeks. Tail‐cuff blood pressure, food/water intake and blood glucose states were monitored daily. 3.A dose dependent decrease in blood glucose was observed. The blood pressure was significantly elevated in rats treated with 9 U/Kg and 7U/Kg doses as compared to other units. However, there was no change in urine output among all the groups. High zinc diet + 7U/Kg and HO‐1 inhibitor ZnDPBG (20mg/kg) treatments produced a significant reduction in blood pressure as compared to 7U/Kg alone. In addition, HO‐1 protein levels in heart and kidney and endogenous CO levels were reduced in high zinc diet + 7U/Kg vs 7U/Kg rats. 4.These results demonstrate that RIIH promotes hypertension and restoration of normal CO levels with a high zinc diet and ZnDPBG reduces the blood pressure. This article is protected by copyright. All rights reserved.
    October 03, 2013   doi: 10.1111/1440-1681.12183   open full text
  • Riluzole reduces arrhythmias and myocardial damage induced by coronary occlusion in anaesthetised pigs.
    S M Weiss, J E Dahlstrom, D A Saint.
    Clinical and Experimental Pharmacology and Physiology. September 24, 2013
    The cardiac persistent sodium current (INaP) presents a novel target for cardiac ischaemic protection. Here we investigated the effects of the INaP blocker riluzole in a pig model of regional myocardial ischaemia. Landrace or Large White pigs were subjected to 3 h of ligation of the left anterior descending coronary artery (LAD). Animals received either saline IV (500 ml/hr) throughout the experiment (CONT: n=7) or riluzole IV (2 mg/kg in 2 ml propylene glycol in 100 ml saline) (RIL: n=7) between 15 minutes and 5 minutes prior to ligation. Arrhythmia score was calculated in 5 min epochs. Myocardial damage was assessed using epicardial image analysis and histological sectioning. In CONT, 7/7 animals developed premature ventricular contractions (PVCs), 7/7 developed non‐sustained arrhythmias, 6/7 developed sustained arrhythmias. Of the sustained arrhythmias, 23/28 instances were initiated by R‐on‐T extrasytoles (extrasystoles within the vulnerable period which can trigger re‐entrant arrhythmias). In RIL, 7/7 animals developed PVCs, 6/7 developed non‐sustained arrhythmias, and only 3/7 developed sustained arrhythmias, of which 2/5 instances were R‐on‐T initiated. RIL treated animals showed less myocardial damage compared to CONT (65% smaller surface area (p=0.008) on gross epicardial inspection, 51% less oedema (p=0.01), 53% less fibre waviness (p=0.029) assessed by H&E staining, and 79% fewer fragmented nuclei (p=0.009) assessed by TUNEL. Riluzole significantly reduced Phase 2 (the period associated with irreversible damage) ischaemic R‐on‐T triggered and non‐R‐on‐T arrhythmias and myocardial damage occurring during the 3 h period of regional ischaemia. This article is protected by copyright. All rights reserved.
    September 24, 2013   doi: 10.1111/1440-1681.12175   open full text
  • Cerebral open flow microperfusion ‐ a new in vivo technique for continuous measurement of substance transport across the intact blood‐brain barrier.
    Thomas Birngruber, Arijit Ghosh, Veronica Perez‐Yarza, Thomas Kroath, Maria Ratzer, Thomas R. Pieber, Frank Sinner.
    Clinical and Experimental Pharmacology and Physiology. September 24, 2013
    The blood‐brain barrier (BBB) limits substance transport to the brain and is therefore the major hurdle to overcome when developing neuroactive drugs. We report herein cerebral open flow microperfusion (cOFM) as a new membrane‐free technique for measuring substance transport across the intact BBB. cOFM is based on a probe that is inserted into the brain causing rupture of the BBB. The BBB is re‐established within 15 days and allows sampling of interstitial brain fluid under physiological conditions. The aim of this proof‐of‐concept study was (i) to determine the time between cOFM probe insertion and BBB re‐establishment; and (ii) to demonstrate the ability of cOFM to sample in the interstitial cerebral fluid with intact BBB. Re‐establishment of the BBB was determined by using Evans Blue (EB). EB is an established marker for BBB intactness as it does not cross the intact BBB. EB levels in the brain tissue indicated that the BBB was healed 11 days after probe insertion. To demonstrate transport across the healed BBB, sodium fluorescein (Naf) was used. Naf is a sensitive, low‐molecular‐weight marker that can cross the intact BBB and can be used to monitor changes in BBB permeability. Significantly increased Naf levels were found in the interstitial fluid when hyperosmolar mannitol (known to open the BBB) was introduced via cOFM, which indicated a partial opening of the BBB surrounding the cOFM probe. We show that cOFM allows monitoring of BBB permeability, which should be useful for measuring pharmacokinetics across the BBB and pharmacodynamics in the brain. This article is protected by copyright. All rights reserved.
    September 24, 2013   doi: 10.1111/1440-1681.12174   open full text
  • Echocardiographic assessment of β‐adrenergic stimulation‐induced heart failure with reduced heart rate in mice.
    Hao Li, Zhi‐Zhen Lu, Chao Chen, Yao Song, Han Xiao, You‐Yi Zhang.
    Clinical and Experimental Pharmacology and Physiology. September 24, 2013
    Chronic injection with the β‐adrenergic receptor (β‐AR) agonist isoproterenol (ISO) has been commonly used as an animal model of β‐AR–induced cardiac remodeling and heart failure. This ISO‐treated model usually shows significantly decreased conscious heart rate (HR). However, the HR in treatment groups is usually adjusted to the same levels by anaesthesia to assess cardiac geometry and function. Here, we reveal a method of echocardiography assessment that represents the true cardiac geometry and function. C57BL/6 mice received 5 mg/kg/day ISO for 12 weeks. Cardiac geometry and function were assessed by high‐resolution echocardiography in vehicle‐ and ISO‐treated mice under conscious or varied anesthetic conditions. Cardiac β‐AR response was decreased in ISO‐treated mice, as evidenced by markedly decreased conscious HR. Vehicle‐ and ISO‐treated mice did not differ in cardiac geometry or function when HR was adjusted to the same level (400 beats/min) in both treatment groups but did differ when using a low (1%) rather than high (1.5% or 2%) isoflurane concentration for HR adjustment. Furthermore, 3‐day ISO withdrawal eliminated the difference in conscious HR between the 2 groups. Also, the groups differed in cardiac geometry and function values regardless of isoflurane concentration. Use of isoflurane to decrease HR of treatment groups to the same level may mask the left‐ventricular dysfunction of ISO‐treated mice. Therefore, we demonstrate that ISO withdrawal eliminates the interference of different basal HR between treatment groups on echocardiography measurements, allowing a more accurate assessment of cardiac pathological and functional changes. This article is protected by copyright. All rights reserved.
    September 24, 2013   doi: 10.1111/1440-1681.12176   open full text
  • Aberrant Rac1–mineralocorticoid receptor pathways in salt‐sensitive hypertension.
    Wakako Kawarazaki, Toshiro Fujita.
    Clinical and Experimental Pharmacology and Physiology. September 24, 2013
    According to Guyton's model, impaired renal sodium excretion plays a key role in the increased salt sensitivity of blood pressure (BP). Several factors contribute to impaired renal sodium excretion, including the sympathetic nervous system, the renin–angiotensin system and aldosterone. Accumulating evidence suggests that abnormalities in aldosterone and its receptor (i.e. the mineralocorticoid receptor (MR)) are involved in the development of salt‐sensitive (SS) hypertension. Patients with metabolic syndrome often exhibit hyperaldosteronism and are susceptible to SS hypertension. Aldosterone secretion from the adrenal glands is not suppressed in obese hypertensive rats fed a high‐salt diet because of the abundant production of adipocyte‐derived aldosterone‐releasing factors, which are independent of the negative feedback regulation of aldosterone secretion by the renin–angiotensin–aldosterone system. Increased plasma aldosterone levels lead to SS hypertension via MR activation in the kidney. Renal MR activity is increased in Dahl salt‐sensitive rats fed a high‐salt diet, despite the appropriate suppression of plasma aldosterone levels. In this rat strain, activation of MR in the distal nephron causes salt‐induced hypertension. This paradoxical response of the MR to salt loading can be attributed to activation of Rac1, a small GTPase. In the presence of aldosterone, activated Rac1 synergistically and directly activates MR in a ligand‐independent manner. Thus, Rac1 activation in the kidney determines the salt sensitivity of BP. Together, the available evidence suggests that the aberrant Rac1–MR pathway plays a key role in the development of SS hypertension. This article is protected by copyright. All rights reserved.
    September 24, 2013   doi: 10.1111/1440-1681.12177   open full text
  • Synthetic peptides from heat‐shock protein 65 inhibit pro‐inflammatory cytokine secretion by peripheral blood mononuclear cells of rheumatoid arthritis patients.
    Jun Zhou, Li‐Ping Wang, Xuan Feng, Dan‐Dan Fan, Wei‐Jin Zang, Bing Wang.
    Clinical and Experimental Pharmacology and Physiology. September 24, 2013
    Rheumatoid arthritis (RA) is a systemic autoimmune disease mediated by T cells. Pro‐inflammatory cytokine plays a critical role in the pathogenesis of RA. The aim of the present study was to investigate the effects of synthetic peptides (HP‐R1, HP‐R2 and HP‐R3), derived from the sequence of 65‐kD mycobacterial heat shock protein (HSP), on proliferation and cytokine secretion by peripheral blood mononuclear cells (PBMCs) of RA patients. PBMCs were obtained from RA patients and collected by Ficoll‐Hypaque density centrifugation. PBMCs were treated with one of three synthetic peptides for 4h, after which the proliferation and cytokine production were determined. The effects of three peptides on proliferation of PBMCs were analyzed by a colorimetric cell proliferation assay (CCK‐8 assay). Cytokine production was measured in culture supernatants using specific ELISA. The three peptides did not significantly affect the proliferation of PBMCs from healthy controls. Cell proliferation of RA patients was inhibited by three peptides. The production of tumor necrosis factor (TNF)‐α was significantly inhibited by three peptides. HP‐R1 and HP‐R2 efficiently suppressed interferon (IFN)‐γ secretion. Unlike the other two HP‐Rs, HP‐R2 increased the IL‐4 level. The production of IL‐10 was not significantly affected by any of the peptides. The present study suggests that the synthetic peptides derived from HSP65 display anti‐proliferative and anti‐inflammatory function and supports the potential use of the synthetic peptides as a therapeutic drug in RA patients. This article is protected by copyright. All rights reserved.
    September 24, 2013   doi: 10.1111/1440-1681.12178   open full text
  • Inhibitory effect of oral contraceptives on CYP2C19 activity is not significant in carriers of the CYP2C19*17 allele.
    Rasmus S. Pedersen, Lene Noehr‐Jensen, Kim Brosen.
    Clinical and Experimental Pharmacology and Physiology. September 23, 2013
    The purpose of the present study was to examine whether cytochrome P450 2C19 (CYP2C19) in carriers of the CYP2C19*17 allele is inhibited in vivo by oral contraceptives (OC). Retrospective CYP2C19 phenotyping according to omeprazole : 5‐OH‐omeprazole molar 3 h plasma metabolic ratios (MR) from a population (n = 222) genotyped as CYP2C19*1/*1, CYP2C19*1/*17 and CYP2C19*17/*17 was analysed. Furthermore, 30 women genotyped as CYP2C19*1/*1 (n = 11), CYP2C19*1/*17 (n = 11) and CYP2C19*17/*17 (n = 8) were prospectively CYP2C19 phenotyped during the administration of OC and again after a minimum 5 days break from OC. We found a significantly higher MR in the CYP2C19*1/*1 genotype group that took OC (n = 48) compared with women who did not take OC (n = 31; geometric mean 1.37 vs 0.83, respectively; P < 0.05). However, in the CYP2C19*1/*17 genotype group, the geometric means of the MR in the 37 women taking OC and the 20 women not taking OC were 0.67 and 0.46, respectively (P > 0.05). In the CYP2C19*1/*1 panel of the prospective cross‐over study, we found a significantly higher MR while women were taking the OC compared with the MR during the OC break (geometric mean 1.21 vs 0.91, respectively; P = 0.0123). However, in the CYP2C19*1/*17 group, the geometric means of the MR with and without OC were 0.77 and 0.65, respectively, compared with 1.05 and 0.79, respectively, in the CYP2C19*17/*17 group (P = 0.20 and 0.17, respectively). In conclusion, we have shown that OC intake inhibits CYP2C19 in homozygous carriers of the CYP2C19 wild type but that the inhibition is not significant in heterozygous and homozygous carriers of the CYP2C19*17 allele.
    September 23, 2013   doi: 10.1111/1440-1681.12153   open full text
  • Differential effects of a gelatinase inhibitor on adipocyte differentiation and adipose tissue development.
    Matthias Van Hul, Dries Bauters, Roger H Lijnen.
    Clinical and Experimental Pharmacology and Physiology. September 23, 2013
    A potential role for the gelatinases in adipocyte differentiation in vitro and adipose tissue development in vivo was investigated using the gelatinase inhibitor tolylsam ((R)‐3‐methyl‐2‐[4‐(3‐p‐tolyl‐[1,2,4]oxadiazol‐5‐yl)‐benzenesulphonylamino]‐butyric acid). Differentiation of murine 3T3‐F442A preadipocytes (12 days after reaching confluence) into mature adipocytes in vitro was promoted in the presence of tolylsam (10–100 μmol/L). De novo development of fat tissue in nude mice injected with preadipocytes and kept on a high‐fat diet was significantly impaired following treatment with tolylsam (100 mg/kg per day for 4 weeks). Adipose tissue development in matrix metalloproteinase (MMP)‐2 deficient mice, kept on a high‐fat diet, was significantly impaired following administration of tolylsam (100 mg/kg per day for 15 weeks). This was associated with markedly enhanced metabolic rate. Treatment of MMP‐2‐deficient mice with tolylsam (100 mg/kg per day, 15 weeks) was associated with the preservation of collagen and a reduction in blood vessel size in adipose tissues in vivo. Furthermore, plasma levels of triglycerides and free fatty acids were reduced by tolylsam treatment of MMP‐2‐deficient mice (100 mg/kg per day, 15 weeks), whereas nutrient adsorption in the intestine was not affected. The results of the present study indicate that tolylsam promotes preadipocyte differentiation in vitro, but impairs adipose tissue development in vivo.
    September 23, 2013   doi: 10.1111/1440-1681.12154   open full text
  • Anandamide modulates the neuroendocrine responses induced by extracellular volume expansion.
    Silvia G Ruginsk, Ernane T Uchoa, Lucila LK Elias, Jose Antunes‐Rodrigues.
    Clinical and Experimental Pharmacology and Physiology. September 23, 2013
    The aim of the present study was to evaluate the effects of intracerebroventricular administration of anandamide (AEA), an inhibitor of fatty acid amide hydrolase activity (URB597) and a CB1 receptor (CB1R) antagonist (AM251) on the homeostatic responses elicited by extracellular volume expansion (EVE) in male adult rats. Pretreatment with AEA (100 ng/4 μL) significantly reduced the effect of hypertonic (H‐) EVE on plasma concentrations of prolactin (PRL), oxytocin (OT) and corticosterone, but not vasopressin (AVP). Administration of URB597 (20 μg/5 μL) alone significantly reduced PRL, OT, AVP and corticosterone in the H‐EVE group. Conversely, URB597 and AEA had no significant effect on basal hormone concentrations. Pretreatment with AM251 (200 ng/2 μL) potentiated OT but did not change AVP plasma levels in the H‐EVE group. Hypertonic EVE significantly increased AVP and OT mRNA expression in the supraoptic nucleus (SON), an effect that was blunted in AEA‐pretreated rats. Pretreatment with AEA did not change the percentage of vasopressinergic or oxytocinergic neurons colocalizing c‐Fos in the SON, but increased nitrate concentrations in the median eminence of animals subjected to H‐EVE. The present data suggest that: (i) vasopressinergic and oxytocinergic neurons may be differentially affected by AEA; (ii) activation of CB1R may restrain the response of the neurohypophyseal system (NHS) to EVE; (iii) the hypothalamic–pituitary–adrenal axis, PRL and the NHS may still be sensitive to AEA after EVE, with these effects probably not dependent on AEA metabolism; and (iv) AEA and nitric oxide could interact in vivo as modulators to directly control stress‐induced responses.
    September 23, 2013   doi: 10.1111/1440-1681.12155   open full text
  • Inhibition of DNA methylation attenuates low‐dose cadmium‐induced cardiac contractile and intracellular Ca2+ anomalies.
    Subat Turdi, Weixia Sun, Yi Tan, Xiaohui Yang, Lu Cai, Jun Ren.
    Clinical and Experimental Pharmacology and Physiology. September 23, 2013
    Cadmium is a human carcinogen with unfavourable health impacts probably associated with its DNA methylation property. Recent data suggest that environmental cadmium exposure is associated with the incidence of myocardial infarction and peripheral arterial disease. Nonetheless, the effect of chronic cadmium exposure on cardiac contractile function remains unknown. The present study was designed to examine the impact of low‐dose cadmium exposure on cardiac contractile function and intracellular Ca2+ homeostasis. Adult male mice were exposed to cadmium for 4 weeks (20 nmol/kg, i.p. every other day for 4 weeks) with or without the DNA methylation inhibitor 5‐aza‐2′‐deoxyctidene (5‐AZA; 0.25 mg/kg, i.p., twice a week for 6 weeks, starting at the same time as cadmium administration). Cardiac contractile and intracellular Ca2+ properties were analysed, including echocardiographic left ventricular parameters, fractional shortening (FS), peak shortening (PS) amplitude, maximal velocity of shortening/relengthening (±dL/dt), time to PS (TPS), time to 90% relengthening (TR90), electrically stimulated increases in intracellular Ca2+ and intracellular Ca2+ decay. Cadmium exposure depressed FS, PS, ±dL/dt and electrically stimulated increases in intracellular Ca2+ without affecting TPS, TR90, intracellular Ca2+ levels or the decay rate. The effects of cadmium were significantly attenuated (PS) or blocked altogether (all other parameters) by 5‐AZA. Cadmium exposure led to overt interstitial fibrosis (collagen deposition), which was mitigated by 5‐AZA treatment. Western blot analysis revealed that cadmium exposure and/or 5‐AZA treatment had no effect on the expression of intercellular adhesion molecule‐1, tumour necrosis factor‐α and cleaved caspase 3, suggesting a relatively minor role of proinflammatory cytokines and apoptosis in the cardiac responses to cadmium and 5‐AZA. Together, our data demonstrate, for the first time, direct cardiac depressant effects following cadmium exposure, which may be rescued by inhibition of DNA methylation.
    September 23, 2013   doi: 10.1111/1440-1681.12158   open full text
  • Is there a role for Day‐to‐day home blood pressure variability in guiding management of hypertension?
    Takayoshi Ohkubo, Anastasia S Mihailidou.
    Clinical and Experimental Pharmacology and Physiology. September 21, 2013
    Although two population‐based studies have demonstrated the prognostic value of day‐to‐day home blood pressure (BP) variability, data are still limited. Thresholds of normality, target levels for treatment, and optimal number and times of home measurement needed for better assessment of day‐to‐day home BP variability also remain to be investigated. Although further prospective studies are required, home BP is a strong and modifiable risk factor and patient compliance should be encouraged. This article is protected by copyright. All rights reserved.
    September 21, 2013   doi: 10.1111/1440-1681.12172   open full text
  • Cholinergic stimulation with pyridostigmine improves autonomic function in infarcted rats.
    Raquel N La Fuente, Bruno Rodrigues, Ivana C Moraes‐Silva, Leandro E Souza, Raquel Sirvente, Cristiano Mostarda, Kátia De Angelis, Pedro P Soares, Silvia Lacchini, Fernanda Consolim‐Colombo, Maria‐Cláudia Irigoyen.
    Clinical and Experimental Pharmacology and Physiology. August 23, 2013
    In the present study we evaluated the effects of short‐term pyridostigmine bromide (0.14 mg/mL) treatment started early after myocardial infarction (MI) on left ventricular (LV) and autonomic functions in rats. Male Wistar rats were divided into control, pyridostigmine, infarcted and infarcted + pyridostigmine‐treated groups. Pyridostigmine was administered in the drinking water, starting immediately after MI or sham operation, for 11 days. Left ventricular function was evaluated indirectly by echocardiography and directly by LV catheterization. Cardiovascular autonomic control was evaluated by baroreflex sensitivity (BRS), heart rate variability (HRV) and pharmacological blockade. All evaluations started after 7 days pyridostigmine treatment and were finalized after 11 days treatment. Pyridostigmine prevented the impairment of +dP/dT and reduced the MI area in infarcted + pyridostigmine compared with infarcted rats (7 ± 3% vs 17 ± 4%, respectively). Mean blood pressure was restored in infarcted + pyridostigmine compared with infarcted rats (103 ± 3 vs 94 ± 3 mmHg, respectively). In addition, compared with the infarcted group, pyridostigmine improved BRS, as evaluated by tachycardic (1.6 ± 0.2 vs 2.5 ± 0.2 b.p.m./mmHg, respectively) and bradycardic (−0.42 ± 0.01 vs −1.9 ± 0.1 b.p.m./mmHg) responses, and reduced the low frequency/high frequency ratio of HRV (0.81 ± 0.11 vs 0.24 ± 0.14, respectively). These improvements are probably associated with increased vagal tone and reduced sympathetic tone in infarcted + pyridostigmine compared with infarcted rats. In conclusion, the data suggest that short‐term pyridostigmine treatment started early after MI can improve BRS, HRV and parasympathetic and sympathetic tone in experimental rats. These data may have potential clinical implications because autonomic markers have prognostic significance after MI.
    August 23, 2013   doi: 10.1111/1440-1681.12121   open full text
  • Exercise training upregulates nitric oxide synthases in the kidney of rats with chronic heart failure.
    Daisuke Ito, Osamu Ito, Nobuyoshi Mori, Pengyu Cao, Chihiro Suda, Yoshikazu Muroya, Kiyotaka Hao, Hiroaki Shimokawa, Masahiro Kohzuki.
    Clinical and Experimental Pharmacology and Physiology. August 23, 2013
    There is an interaction between heart and kidney diseases, which is a condition termed cardiorenal syndrome. Exercise training has cardioprotective effects, involving upregulation of endothelial (e) nitric oxide synthase (NOS) in the cardiovascular system. However, the effects of exercise training on NOS in the kidney with heart disease are unknown. The aim of the present study was to investigate whether exercise training upregulates NOS in the kidney, left ventricle and aorta of rats with chronic heart failure (CHF). Male Sprague‐Dawley rats underwent left coronary artery ligation (LCAL) to induce CHF and were randomly assigned to sedentary or treadmill exercise groups 4 weeks after LCAL. Three days after exercising for 4 weeks, urine samples were collected for 24 h and blood samples were collected following decapitation. Nitric oxide synthase activity and protein expression were examined. Significant interactions between CHF and exercise training were observed on parameters of cardiac and renal function. Exercise training improved cardiac function, decreased plasma B‐type natriuretic peptide levels, decreased urinary albumin excretion and increased creatinine clearance in CHF rats. Nitric oxide synthase activity, eNOS expression and neuronal (n) NOS expression were significantly decreased in the left ventricle and kidney of CHF rats. Exercise training significantly increased NOS activity and eNOS and nNOS expression. Upregulation of NOS in the kidney and left ventricle may contribute, in part, to the renal and cardiac protective effects of exercise training in cardiorenal syndrome in CHF rats.
    August 23, 2013   doi: 10.1111/1440-1681.12130   open full text
  • Oxidative stress inhibits adhesion and transendothelial migration, and induces apoptosis and senescence of induced pluripotent stem cells.
    Yi Wu, Xueqing Zhang, Xueling Kang, Ning Li, Rong Wang, Tiantian Hu, Meng Xiang, Xinhong Wang, Wenjun Yuan, Alex Chen, Dan Meng, Sifeng Chen.
    Clinical and Experimental Pharmacology and Physiology. August 23, 2013
    Oxidative stress caused by cellular accumulation of reactive oxygen species (ROS) is a major contributor to disease and cell death. However, how induced pluripotent stem cells (iPSC) respond to different levels of oxidative stress is largely unknown. Here, we investigated the effect of H2O2‐induced oxidative stress on iPSC function in vitro. Mouse iPSC were treated with H2O2 (25–100 μmol/L). IPSC adhesion, migration, viability, apoptosis and senescence were analysed. Expression of adhesion‐related genes, stress defence genes, and osteoblast‐ and adipocyte‐associated genes were determined by reverse transcription polymerase chain reaction. The present study found that H2O2 (25–100 μmol/L) decreased iPSC adhesion to matrix proteins and endothelial cells, and downregulated gene expression levels of adhesion‐related molecules, such as integrin alpha 7, cadherin 1 and 5, melanoma cell adhesion molecule, vascular cell adhesion molecule 1, and monocyte chemoattractant protein‐1. H2O2 (100 μmol/L) decreased iPSC viability and inhibited the capacity of iPSC migration and transendothelial migration. iPSC were sensitive to H2O2‐induced G2/M arrest, senescence and apoptosis when exposed to H2O2 at concentrations above 25 μmol/L. H2O2 increased the expression of stress defence genes, including catalase, cytochrome B alpha, lactoperoxidase and thioredoxin domain containing 2. H2O2 upregulated the expression of osteoblast‐ and adipocyte‐associated genes in iPSC during their differentiation; however, short‐term H2O2‐induced oxidative stress did not affect the protein expression of the pluripotency markers, octamer‐binding transcription factor 4 and sex‐determining region Y‐box 2. The present results suggest that iPSC are sensitive to H2O2 toxicity, and inhibition of oxidative stress might be a strategy for improving their functions.
    August 23, 2013   doi: 10.1111/1440-1681.12141   open full text
  • Icariin attenuates high glucose‐induced type IV collagen and fibronectin accumulation in glomerular mesangial cells by inhibiting transforming growth factor‐β production and signalling through G protein‐coupled oestrogen receptor 1.
    Yi‐Chen Li, Xuan‐Sheng Ding, Hui‐Mei Li, Cheng Zhang.
    Clinical and Experimental Pharmacology and Physiology. August 23, 2013
    Icariin has been shown to attenuate diabetic nephropathy in rats by decreasing transforming growth factor‐β (TGF‐β) and type IV collagen expression, but its mode of action in glomerular mesangial cells is uncertain. The present study aimed to investigate the effect of icariin on excess mesangial type IV collagen and fibronectin accumulation induced by high glucose, and to determine the mechanism underlying its protective effects. Under high‐glucose conditions, icariin diminished type IV collagen and fibronectin accumulation, as well as TGF‐β production in human and rat mesangial cells. Mesangial cells treated with icariin after TGF‐β1 exposure expressed less type IV collagen and fibronectin than those without icariin treatment, suggesting inhibition by icariin of TGF‐β1 downstream pathways. On TGF‐β1 stimulation, icariin inhibited TGF‐β canonical Smad signalling and extracellular signal‐regulated kinase (ERK)1/2 signalling by decreasing Smad2/3 and ERK1/2 phosphorylation in a dose‐dependent manner. U0126, which blocked the ERK1/2 pathway, exerted an additive effect on the icariin suppression of type IV collagen and fibronectin expression, enhancing the beneficial effects of icariin. The G protein‐coupled oestrogen receptor 1 (GPER) antagonist, G‐15, abolished the icariin‐induced inhibition of type IV collagen, and fibronectin overproduction and TGF‐β signalling. Treatment of cells with fulvestrant, a downregulator of the oestrogen receptor, enhanced the action of icariin. In conclusion, icariin decreased type IV collagen and fibronectin accumulation induced by high glucose in mesangial cells by inhibiting TGF‐β production, as well as Smad and ERK signalling in a GPER‐dependent manner.
    August 23, 2013   doi: 10.1111/1440-1681.12143   open full text
  • Pioglitazone alleviates the mitochondrial apoptotic pathway and mito‐oxidative damage in the d‐galactose‐induced mouse model.
    Atish Prakash, Anil Kumar.
    Clinical and Experimental Pharmacology and Physiology. August 23, 2013
    Chronic injection of d‐galactose can cause gradual deterioration in learning and memory capacity, and activates oxidative stress, mitochondrial dysfunction and apoptotic cell death in the brain of mice. Thus, it serves as an animal model of ageing. Recent evidence has shown that mild cognitive impairment in humans might be alleviated by treatment with piogliatzone (peroxisome proliferator‐activated receptor gamma (PPARγ) agonists). To continue exploring the effects of piogliatzone in this model, we focused on behavioural alteration, oxidative damage, mitochondrial dysfunction and apoptosis in d‐galactose‐induced mice. The ageing model was established by administration of d‐galactose (100 mg/kg) for 6 weeks. Pioglitazone (10 and 30 mg/kg) and bisphenol A diglycidyl ether (15 mg/kg) were given daily to d‐galactose‐induced senescent mice. The cognitive behaviour of mice was monitored using the Morris water maze. The anti‐oxidant status and apoptotic activity in the ageing mice was measured by determining mito‐oxidative parameters and caspase‐3 activity in brain tissue. Systemic administration of d‐galactose significantly increased behavioural alterations, biochemical parameters, mitochondrial enzymes, and activations of caspase‐3 and acetylcholinesterase enzyme activity as compared with the control group. Piogliatzone treatment significantly improved behavioural abnormalities, biochemical, cellular alterations, and attenuated the caspase‐3 and acetylcholinesterase enzyme activity as compared with the control. Furthermore, pretreatment of BADGE (PPARγ antagonist) with pioglitazone reversed the protective effect of pioglitazone in d‐galactose‐induced mice. The present study highlights the protective effects of pioglitzone against d‐galactose‐induced memory dysfunction, mito‐oxidative damage and apoptosis through activation of PPARγ receptors. These findings suggest that pioglitazone might be helpful for the prevention or alleviation of ageing.
    August 23, 2013   doi: 10.1111/1440-1681.12144   open full text
  • Supplementation of the maternal diet during pregnancy with chocolate and fructose interacts with the high‐fat diet of the young to facilitate the onset of metabolic disorders in rat offspring.
    Zhi‐Yun Zhang, Yun‐Bin Dai, Hao‐Nan Wang, Ming‐Wei Wang.
    Clinical and Experimental Pharmacology and Physiology. August 23, 2013
    Obesity and non‐alcoholic fatty liver disease are the most common metabolic disorders in society today. Previously, we found that supplementing the maternal diet during pregnancy with chocolate and fructose has negative effects on the well‐being of the offspring that were ameliorated if the offspring were fed a normal diet during postnatal life. In the present study, we investigated whether feeding offspring a high‐fat diet would augment the maternal programming effects and whether extra protein supply can correct the low birth weight resulting from the chocolate‐supplemented maternal diet. Pregnant Sprague‐Dawley rats were divided into three groups and fed either standard chow (normal nutrition; NN), chocolate‐ and fructose‐supplemented standard chow with casein sodium (overnutrition; ON) or the supplemented standard chow without casein sodium (malnutrition; MN) throughout pregnancy. Male offspring were weaned on either standard or high‐fat chow. Dams in the MN group exhibited moderate weight gain, consumed 50% less protein (P < 0.001) but more carbohydrates during gestation and delivered pups with a 12% lower birth weight (P < 0.05) than pups in the NN group, results that are consistent with previous findings. When fed on a high‐fat diet after birth, pups from dams in the MN group (MNHD) had 30% more body fat (P = 0.023) and liver triglyceride (TG) levels that were double (P < 0.01) those in offspring in the other groups, leading to fatty livers in these offspring at 14 weeks of age. Hepatic expression of the PPARα, ApoB100, MTTP, CPT1 and SREBP1c genes was significantly downregulated in the MNHD group (P < 0.05 for all), indicating changes in lipid metabolism. Although dams in the ON group exhibited marked gestational weight gain (P < 0.01), they gave birth to normal weight pups that only manifested mild increases in body fat and liver TG content (P < 0.05), without significant changes in the expression of most genes when fed with the high‐fat diet. The results suggest that the extra protein supply in the form of casein sodium was able to correct some negative programming effects of the chocolate and fructose supplementation of the maternal diet, which, in conjunction with a high‐fat diet in the offspring, may facilitate the onset of metabolic disorders, with impaired liver gene expression possibly a key contributor.
    August 23, 2013   doi: 10.1111/1440-1681.12147   open full text
  • Serum amyloid A stimulates cultured endothelial cells to migrate and proliferate: inhibition by the multikinase inhibitor BIBF1120.
    Xiaoping Cai, S Ben Freedman, Paul K Witting.
    Clinical and Experimental Pharmacology and Physiology. August 23, 2013
    In the present study, we tested whether serum amyloid A (SAA) protein, an established biomarker of inflammation, also plays a role in stimulating neovascularization. To evaluate this possibility, human carotid artery endothelial (HCtAE) cells were cultured and cellular migration and the proinflammatory and/or thrombotic activity of SAA (0, 1 or 10 μg/mL) on vascular endothelial cells was verified by determining gene regulation relative to control (in the absence of SAA). Exposure of HCtAE cells to SAA increased expression of the transcription factor nuclear factor‐κB (NFKB), tumour necrosis factor (TNF) and pro‐coagulative tissue factor (F3), and stimulated phosphorylation of the P65 subunit of the NFKB complex. Enhanced production of TNF and NFKB was paralleled by increased vascular endothelial growth factor (VEGF) mRNA and protein expression, as demonstrated by quantitative polymerase chain reaction, western blotting and ELISA. Administration of 10 μg/mL SAA enhanced endothelial cell migration (1.6‐fold vs control), stimulated regrowth of HCtAE cells after mechanical injury (˜1.2‐fold vs control) and increased endothelial tube formation relative to control after 6 h. The SAA‐mediated enhancement of endothelial cell migration, proliferation and tube formation were markedly inhibited by pretreatment of HCtAE cells with the multi‐angiokinase receptor inhibitor BIBF1120 (100 nmol/L), although SAA‐stimulated gene responses for F3 and NFKB were unaffected by 100 nmol/L BIBF1120 pretreatment. Overall, BIBF1120 inhibited the pro‐angiogenic activity of SAA on vascular endothelial cells in this experimental model of inflammation.
    August 23, 2013   doi: 10.1111/1440-1681.12148   open full text
  • Grb10 Inhibits Glucose‐stimulated Insulin Release from Pancreatic β‐Cells Associated with Suppression of Insulin/IGF‐1 Signaling Pathway.
    Ling Li, Xiaowen Li, Yunxia Zhu, Mingliang Zhang, Dechao Yin, Junxi Lu, Feng Liu, Chen Wang, Weiping Jia.
    Clinical and Experimental Pharmacology and Physiology. August 13, 2013
    Growth receptor binding protein 10 (Grb10) is an adaptor protein that interacts with the insulin receptor and insulin like growth factor‐1 receptor. Overexpression of Grb10 in muscle cells and adipocytes inhibits insulin signaling and transgenic mice overexpressing Grb10 presented with impaired glucose tolerance. However, the roles of Grb10 in β‐cells remain unknown. Aim of the study was to explore the effect of Grb10 on β‐cell function. Grb10 protein expression was detected in islets or INS 832/13 cells with western blot. The effects of Grb10 on glucose‐stimulated insulin secretion (GSIS) and the insulin/IGF‐1 signaling pathway in islets were investigated in rat islets and/or dispersed islet cells with Grb10 overexpresion by adenovirus transfection. Grb10 is expressed in both human and rat pancreas. Its expression is increased in islets isolated from rats fed with high fat plus high sugar diet compared with those isolated from rats fed with chow diet, and in INS 832/13 cells exposed to high level of glucose (20 mM), palmitate (1 mM), interleukin‐1β (50 U/ml). Overexpression of Grb10 in INS 832/13 cells or rat islets impaired GSIS compared with their respective controls (all P < 0.05). Moreover, the inhibition of GSIS by Grb10 overexpression was associated with decrease of insulin‐ or IGF‐1‐induced Akt and ERK1/2 phosphorylation. Our study demonstrates that Grb10 is an important negative regulator of insulin/IGF‐1 signaling in pancreatic β‐cells and a potential target to improve β‐cell function. This article is protected by copyright. All rights reserved.
    August 13, 2013   doi: 10.1111/1440-1681.12160   open full text
  • Vascular effects of aldosterone: sorting out the receptors and the ligands.
    Ross D. Feldman, Robert Gros.
    Clinical and Experimental Pharmacology and Physiology. August 01, 2013
    Aldosterone has actions far beyond its role as a renal regulator of sodium reabsorption and broader mechanisms of action than simply a transcriptional regulator. Aldosterone has a number of vascular effects including regulation of vascular reactivity and vascular growth/development. Aldosterone‐mediated effects on vascular reactivity reflect a balance between its endothelial‐dependent vasodilator effects and its direct smooth muscle vasoconstrictor effects. The endothelial vasodilator effects of aldosterone are mediated by phosphoinositide 3 kinase (PI3K)‐dependent nitric oxide synthase (NOS) activation. GPER is a recently recognized G protein coupled receptor (GPCR) that is activated by steroid hormones. It was first recognized as the GPCR mediating the rapid effects of estrogens. GPER activation also mediates at least some of aldosterone's vascular effects in smooth muscle and in endothelial cells. In vascular endothelial cells aldosterone activation of GPER mediates vasodilation. In contrast endothelial MR activation has been linked to enhanced vasoconstrictor and/or impaired vasodilator responses. This article is protected by copyright. All rights reserved.
    August 01, 2013   doi: 10.1111/1440-1681.12157   open full text
  • Mineralocorticoid receptor and cardiac arrhythmia.
    Basile Gravez, Antoine Tarjus, Frederic Jaisser.
    Clinical and Experimental Pharmacology and Physiology. July 29, 2013
    Mineralocorticoid receptor (MR) activation has been shown to play a deleterious role in the development of heart disease in studies using specific MR antagonists (spironolactone, eplerenone) in experimental models and patients. Pharmacological MR blockade attenuates the transition to heart failure (HF) in models of systolic left ventricular dysfunction and myocardial infarction and of diastolic dysfunction, in rats and mice. In humans, MR antagonism is highly beneficial in patients with mild or advanced HF and post‐infarct HF. The consequences of aldosterone and MR activation for cardiac arrhythmia and its prevention/correction by MR antagonists are often underestimated. MR activation modulates cardiac electrical activity, causing atrial and ventricular arrhythmias. A pro‐arrhythmogenic effect of aldosterone (possibly partly dependent on fibrosis) has been suggested by several studies. Cardiac MR activation has important consequences for the control of cellular calcium homeostasis, action potential lengthening, the modulation of calcium transients and sarcoplasmic reticulum diastolic leaks, resulting in the promotion of rhythm disorders. Aldosterone/MR activation (in both cardiomyocytes and coronary vessels) results in vascular dysfunction, and also contributes to pro‐arrhythmogenic conditions. Taken together, these pro‐arrhythmic effects of aldosterone/MR may explain the highly beneficial effect of MR antagonism, decreasing the incidence of sudden death, observed in the RALES and EPHESUS studies. This article is protected by copyright. All rights reserved.
    July 29, 2013   doi: 10.1111/1440-1681.12156   open full text
  • Elevated vascular γ‐butyrobetaine levels attenuate the development of high glucose‐induced endothelial dysfunction.
    Reinis Vilskersts, Olga Zharkova‐Malkova, Rudolfs Mezhapuke, Solveiga Grinberga, Helena Cirule, Maija Dambrova.
    Clinical and Experimental Pharmacology and Physiology. July 29, 2013
    The aim of the present study was to investigate the effects of vascular tissue levels of l‐carnitine and its precursor, γ‐butyrobetaine (GBB), on the development of endothelial dysfunction induced by 5 μmol/L lysophosphatidylcholine (LPC), 10 mmol/L triglycerides (TG) or a high glucose concentration (44 mmol/L). Changes in vascular tissue levels of l‐carnitine and GBB were induced by administration of l‐carnitine (100 mg/kg), mildronate (100 mg/kg; an inhibitor of l‐carnitine synthesis) or their combination to male Wistar rats for 2 weeks. Treatment with l‐carnitine elevated vascular tissue levels of l‐carnitine, whereas administration of mildronate reduced l‐carnitine levels and increased GBB levels. Experimental animals that received the combination of both drugs showed elevated tissue levels of GBB. The results from organ bath experiments demonstrated that increased GBB levels with preserved l‐carnitine content in vascular tissues attenuated the development of endothelial dysfunction induced by high glucose. However, changes in vascular tissue l‐carnitine and GBB levels had no impact on endothelial dysfunction induced by TG or LPC. The results demonstrate that increased levels of GBB with preserved l‐carnitine content in vascular tissue attenuate the development of endothelial dysfunction induced by high glucose concentrations.
    July 29, 2013   doi: 10.1111/1440-1681.12127   open full text
  • Physiological mechanisms for the increase in renal solute‐free water clearance by a glucagon‐like peptide‐1 mimetic.
    Anna V Kutina, Anna S Marina, Elena I Shakhmatova, Yury V Natochin.
    Clinical and Experimental Pharmacology and Physiology. July 29, 2013
    The aim of the present study was to clarify the mechanisms mediating the effect of a glucagon‐like peptide‐1 (GLP‐1) mimetic on solute‐free water excretion in rats. The GLP‐1 mimetic exenatide (0.05–5.0 nmol/kg, i.m.), alone and in combination with either a vasopressin V2 receptor antagonist (15 nmol/kg, i.p.) or vasopressin (0.01 nmol/kg, i.m.), was injected into control and water‐loaded (water 10–50 mL/kg, p.o., or 50 mL/kg of 0.6% NaCl, i.p.) rats to evaluate the role of collecting duct water permeability in the hydrouretic effect. Urinary prostaglandin (PG) E2 excretion and the effects of diclofenac (5 mg/kg, i.m.) and GLP‐1 receptor antagonist (0.15 μmol/kg, i.p.) on exenatide action were assessed. The hydrouretic effect of exenatide was equivalent following oral or intraperitoneal water loading, and was proportional to the volume of water administered. Injection of exenatide, under conditions of a maximal decrease in collecting duct water permeability (V2 receptor antagonist administration in water‐loaded rats), additionally stimulated solute‐free water formation. The GLP‐1 receptor antagonist weakened the hydrouretic action of exenatide. Urinary PGE2 excretion increased following water loading (47 ± 6 vs 24 ± 4 ng/kg over a 30 min period) and was enhanced as a result of additional exenatide injection (69 ± 10 ng/kg). Diclofenac and vasopressin delayed the hydrouretic effect of exenatide. The effect of exenatide on solute‐free water clearance in water‐loaded rats is presumably mediated by stimulation of PGE2 secretion and reinforcement of tubular fluid influx from the proximal tubule to the distal segment of the nephron and collecting duct.
    July 29, 2013   doi: 10.1111/1440-1681.12119   open full text
  • Efffect of the ABCC3 –211C/T polymorphism on clopidogrel responsiveness in patients with percutaneous coronary intervention.
    Jian‐Jun Zou, Hong‐Wei Fan, Shao‐Liang Chen, Jie Tan, Bang‐Shun He, Hong‐Guang Xie.
    Clinical and Experimental Pharmacology and Physiology. July 29, 2013
    Multidrug resistance protein 3 (MPR3), encoded by the ATP‐binding cassette, subfamily C (CFTR/MRP), member 3 (ABCC3) gene, functions as an important drug efflux transporter. The ABCC3 –211C/T polymorphism is associated with decreased MRP3 mRNA expression, and low MRP3 mRNA expression is associated with increased clopidogrel response in patients. The aim of the present study was to determine whether the –211C/T polymorphism is associated with altered antiplatelet effects and clinical outcomes in clopidogrel‐treated patients. A subcohort of 249 patients not carrying the CYP2C19*2, *3 or *17 variant was identified from a total of 617 consecutive clopidogrel‐treated patients undergoing percutaneous coronary intervention and then categorized into three groups on the basis of their ABCC3 –211C/T genotype. Baseline data, clinical characteristics and DNA samples were collected for all patients. Light transmittance aggregometry was used to determine ADP‐induced maximum platelet aggregation (MPA) in blood samples obtained from patients on Day 3 after starting daily clopidogrel maintenance doses. Genotyping of CYP2C19*2, *3 and *17 variants and the ABCC3 –211C/T polymorphism was performed using matrix‐assisted laser desorption ionization time‐of‐flight (MALDI‐TOF) mass spectrometry. The primary clinical end‐point was a definite stent thrombosis (ST) episode, whereas secondary end‐points were other major adverse cardiovascular events within 12 months after stenting. There were no differences in MPA values according to ABCC3 –211C/T genotype. A multiple linear regression model revealed that the ABCC3 –211C/T polymorphism was not independently associated with ADP‐induced MPA measurements; a multiple logistic regression model revealed that carrying the ABCC3 –211C allele was not associated with the risk of developing an ST event in clopidogrel‐treated patients not harbouring CYP2C19*2, *3 and *17 variants. In conclusion, the ABCC3 –211C/T polymorphism seems not to be associated with altered antiplatelet effects and clinical outcomes in clopidogrel‐treated patients.
    July 29, 2013   doi: 10.1111/1440-1681.12118   open full text
  • Protective arms of the renin–angiotensin‐system in neurological disease.
    Colin Sumners, Masatsugu Horiuchi, Robert E Widdop, Claudia McCarthy, Thomas Unger, Ulrike M Steckelings.
    Clinical and Experimental Pharmacology and Physiology. July 29, 2013
     In recent years it has been firmly established that apart from the classic renin–angiotensin system (RAS) comprising angiotensin (Ang) II, angiotensin converting enzyme (ACE; responsible for AngII generation) and the angiotensin AT1 receptor (AT1R), there also exist protective arms of the RAS that comprise the angiotensin AT2 receptor (AT2R), Ang‐(1–7), ACE2 (mainly responsible for Ang‐(1–7) synthesis) and Mas, the receptor for Ang‐(1–7).  Stimulation of AT2R promotes neuronal differentiation, neurite outgrowth and axonal regeneration, which results in an acceleration of repair and improved functional outcome after injury of peripheral nerves or the spinal cord.  Stimulation of AT2R and the receptor Mas has been shown to reduce infarct size and ameliorate neurological deficits in various animal models of stroke. The underlying mechanisms of action are comprised of activation of direct neurotrophic, anti‐inflammatory and anti‐oxidant pathways, as well as the augmentation of cerebral blood flow.  Cognitive function is improved by AT2R stimulation due, at least in part, to increased cerebral blood flow. There is indirect evidence that Ang‐(1–7) could also play a role in protection against cognitive decline, but studies confirming this have not yet been published.  In view of the data reviewed in this article, it can be assumed that the protective arms of the RAS are putative targets in the treatment of neurological diseases, which involve tissue damage or cognitive impairment.
    July 29, 2013   doi: 10.1111/1440-1681.12137   open full text
  • Brain and retinal microglia in health and disease: An unrecognized target of the renin–angiotensin system.
    Claudia A McCarthy, Robert E Widdop, Devy Deliyanti, Jennifer L Wilkinson‐Berka.
    Clinical and Experimental Pharmacology and Physiology. July 29, 2013
    Microglia are the resident immune cells within the brain and retina, commonly known as the macrophages of the central nervous system (CNS). Microglia survey the surrounding milieu to eliminate invading microbes, clear cellular debris and enforce programmed cell death by removing apoptotic cells. Complementary to their ‘house‐keeping’ role, microglia are capable of releasing brain‐derived neurotrophic factor (BDNF), as well as various anti‐inflammatory cytokines that sustain and support neuronal survival. Although microglia are essential for maintaining a healthy CNS, paradoxically they may undergo phenotypic changes to influence numerous neurodegenerative disorders, including Parkinson's disease and Alzheimer's disease. Understanding the underlying mechanisms that determine whether microglia are supportive or toxic could elucidate novel and more effective therapeutic targets to treat an array of neurological and retinal diseases. Although relatively little is known about the influences that evoke phenotypic changes in the microglial population, there is accumulating evidence illustrating an interaction with the renin‐angiotensin system (RAS). The angiotensin AT1 and AT2 receptors may have differential roles in mediating the activity of microglia. Understanding the actions of these angiotensin receptors will be important in defining whether microglia are an important therapeutic target for RAS blockade in brain and ocular diseases.
    July 29, 2013   doi: 10.1111/1440-1681.12099   open full text
  • Do intravenous and subcutaneous angiotensin II increase blood pressure by different mechanisms?
    Duncan J Campbell.
    Clinical and Experimental Pharmacology and Physiology. July 29, 2013
     Angiotensin (Ang) II plays a key role in blood pressure regulation. Mechanisms of the pressor effect of chronic intravenous AngII administration include vasoconstriction, stimulation of the sympathetic nervous system and aldosterone production, as well as direct effects on renal excretion of sodium and water.  Chronic AngII administration by subcutaneous minipump at doses higher than required to increase blood pressure by the intravenous route has identified additional pressor mechanisms, including the immune system, cytokines and matrix metalloproteinases. However, pressor doses of subcutaneous AngII may exceed the angiotensinogen synthesis rate and produce inflammation, fibrosis and necrosis of skin overlying the minipump.  Evidence that chronic subcutaneous and intravenous AngII increase blood pressure by different mechanisms includes the prevention of the pressor effects of subcutaneous, but not intravenous, AngII by angiotensin‐converting enzyme inhibition. Furthermore, low doses of subcutaneous AngII reduce blood pressure of female, but not male, rodents and higher doses are less pressor in females than in males, whereas intravenous AngII is equally pressor in males and females.  Pressor doses of chronic subcutaneous AngII produce greater weight loss, anorexia and reduced kidney weight and cause greater vascular, cardiac and renal pathology than equally pressor doses of chronic intravenous AngII.  The different effects of chronic intravenous and subcutaneous AngII suggest that these two models of hypertension give different information and may differ in their relevance to blood pressure regulation in physiological and pathological states such as hypertension in humans.
    July 29, 2013   doi: 10.1111/1440-1681.12085   open full text
  • Emerging markers in cardiovascular disease: Where does angiotensin‐converting enzyme 2 fit in?
    Sheila K Patel, Elena Velkoska, Louise M Burrell.
    Clinical and Experimental Pharmacology and Physiology. July 29, 2013
    The renin–angiotensin system plays a major role in the pathophysiology of cardiovascular disease (CVD). The enzyme angiotensin‐converting enzyme (ACE) converts angiotensin (Ang) I into the vasoconstrictor AngII and was thought, until recently, to be the main effector of the system. The enzyme ACE2, discovered in 2000, can counterbalance the effects of ACE through degradation of AngII and generation of Ang‐(1–7). Angiotensin‐converting enzyme 2 is abundantly expressed in the heart and localized to the endothelial cells of coronary vessels and smooth muscle cells. Its catalytically active ectodomain undergoes shedding, resulting in ACE2 in the circulation. There are 10 studies to date that have measured circulating ACE2 activity in humans, including in healthy subjects and those with heart failure, Type 1 diabetes, implantable cardioverter/defibrillator, elderly subjects undergoing emergency orthopaedic surgery and kidney transplant patients. The results suggest that circulating ACE2 activity may be a marker of CVD, with low levels in healthy individuals and increased levels in those with cardiovascular risk factors or disease. Whether increased plasma ACE2 activity reflects increased synthesis from tissue ACE2 mRNA or increased shedding of tissue ACE2 remains to be determined. Angiotensin‐converting enzyme 2 is located on the X‐chromosome and circulating ACE2 levels are higher in men than in women. Large clinical studies in CVD are needed to more precisely clarify the role of ACE2 as a biomarker of CVD, determine the prognostic significance of circulating ACE2 activity and assess whether the measurement of ACE2 will improve CVD risk prediction.
    July 29, 2013   doi: 10.1111/1440-1681.12069   open full text
  • Unmasking the potential of the angiotensin AT2 receptor as a therapeutic target in hypertension in men and women: What we know and what we still need to find out.
    Lucinda M Hilliard, Katrina M Mirabito, Kate M Denton.
    Clinical and Experimental Pharmacology and Physiology. July 29, 2013
    Major sex differences exist in the development and progression of hypertension and cardiovascular disease. Prior to menopause, women have lower arterial pressure and, furthermore, are protected from hypertension and cardiovascular disease relative to age‐matched men. However, after menopause this cardiovascular protection in women is lost. These sex differences have been linked to sexual dimorphism in the physiological mechanisms that regulate arterial pressure, including the renin–angiotensin system (RAS), which can also impact on the male and female response to different therapeutic approaches. This suggests that antihypertensive regimens need to be tailored according to sex. Newly discovered components of the RAS have emerged in recent years, allowing us to look beyond the classical RAS for novel therapeutic targets for hypertension. In this context, it is now well established that the angiotensin AT2 receptor (AT2R) elicits depressor and natriuretic effects and that these effects are greater in females due to enhanced AT2R levels modulated by oestrogen. In light of knowledge that AT2R expression is regulated by oestrogen and that the prevalence of hypertension and cardiovascular risk is greater in women after menopause, AT2R agonist therapy may represent an innovative therapeutic approach to treat hypertension. Consequently, understanding how ageing and changes in the sex hormone balance influence the RAS is vital if we are to evaluate the potential of the AT2R as a therapeutic target in women and also in men.
    July 29, 2013   doi: 10.1111/1440-1681.12067   open full text
  • Antifibrotic peptide N‐acetyl‐Ser‐Asp‐Lys‐Pro (Ac‐SDKP): Opportunities for angiotensin‐converting enzyme inhibitor design.
    Ross G Douglas, Mario R Ehlers, Edward D Sturrock.
    Clinical and Experimental Pharmacology and Physiology. July 29, 2013
     The renin–angiotensin system (RAS) is central to regulation of blood pressure and electrolyte homeostasis.  Angiotensin‐converting enzyme (ACE), a key protease in the RAS, has a range of substrates, including N‐acetyl‐Ser‐Asp‐Lys‐Pro (Ac‐SDKP). The peptide Ac‐SDKP is cleared almost exclusively by ACE, and specifically by the N‐domain active site of this enzyme.  N‐Acetyl‐Ser‐Asp‐Lys‐Pro is a negative regulator of haematopoietic stem cell differentiation and is a potent antifibrotic agent. In this review, the physiological actions of Ac‐SDKP are presented, together with the potential clinical usefulness of raising Ac‐SDKP levels. This emphasizes the possible opportunity of N‐domain‐selective ACE inhibitors or ACE‐resistant Ac‐SDKP analogues for the treatment of fibrosis.
    July 29, 2013   doi: 10.1111/1440-1681.12062   open full text
  • Role of angiotensin AT2 receptors in natriuresis: Intrarenal mechanisms and therapeutic potential.
    Robert M Carey, Shetal H Padia.
    Clinical and Experimental Pharmacology and Physiology. July 29, 2013
    The renin–angiotensin system is a coordinated hormonal cascade critical for the regulation of blood pressure (BP) and kidney function. Angiotensin (Ang) II, the major angiotensin effector peptide, binds to two major receptors, namely AT1 and AT2 receptors. The AT1 receptors engender antinatriuresis and raise BP, whereas AT2 receptors oppose these effects, inducing natriuresis and reducing BP. There is high AT2 receptor expression in the adult kidney, especially in the proximal tubule. In AT2 receptor‐null mice, long‐term AngII infusion results in pressor and antinatriuretic hypersensivivity compared with responses in wild‐type mice. The major endogenous receptor ligand for AT2 receptor‐mediated natriuretic responses appears to be des‐aspartyl1‐AngII (AngIII) instead of AngII. Recent studies have demonstrated that AngII requires metabolism to AngIII by aminopeptidase A to induce natriuresis and that inhibition of aminopeptidase N increases intrarenal AngIII and augments AngIII‐induced natriuresis. The renal dopaminergic system is another important natriuretic pathway. Renal proximal tubule the D1 and D5 receptor subtypes (D1‐like receptors (D1LIKER)) control approximately 50% of basal sodium excretion. Recently, we have found that natriuresis induced by proximal tubule D1LIKER requires AT2 receptor activation and that D1LIKER stimulation induces recruitment of AT2 receptors to the apical plasma membrane via a cAMP‐dependent mechanism. Initial studies using the potent AT2 receptor non‐peptide agonist Compound 21 demonstrate natriuresis in both the presence and absence of AT1 receptor blockade, indicating the therapeutic potential of this compound in fluid‐retaining states and hypertension.
    July 29, 2013   doi: 10.1111/1440-1681.12059   open full text
  • Introduction.
    Duncan J Campbell.
    Clinical and Experimental Pharmacology and Physiology. July 29, 2013
    There is no abstract available for this paper.
    July 29, 2013   doi: 10.1111/1440-1681.12134   open full text
  • Regulation of Aldosterone Biosynthesis by the Kir3.4 (KCNJ5) Potassium Channel.
    Carolina Velarde‐Miranda, Elise P. Gomez‐Sanchez, Celso E. Gomez‐Sanchez.
    Clinical and Experimental Pharmacology and Physiology. July 05, 2013
    1.The G‐protein‐activated inwardly rectifying potassium channel Kir3.4 is expressed in the zona glomerulosa cell membrane and transports potassium out of the cell. 2.Angiotensin II stimulation of aldosterone secretion is mediated in part by suppression of the transcription of KCNJ5, the gene coding for Kir3.4, and blocking channel activity. This results in membrane depolarization, mobilization of intracellular calcium, activation of the calcium‐calmodulin pathway, and increasing gene transcription of steroidogenic enzymes required for aldosterone secretion. 3.In 40‐60% of aldosterone‐producing adenomas there is a somatic mutation in the region of the KCNJ5 gene that codes for the selectivity filter that decreases potassium selectivity, allowing sodium to leak into the cells, thus depolarizing the membrane and initiating events that result in increased aldosterone synthesis. 4.The mechanism by which mutated KCNJ5 induces cell proliferation and adenoma formation remains unclear. This article is protected by copyright. All rights reserved.
    July 05, 2013   doi: 10.1111/1440-1681.12151   open full text
  • Replacing Manual Sphygmomanometers with Automated Blood Pressure Measurement in Routine Clinical Practice.
    Martin G. Myers.
    Clinical and Experimental Pharmacology and Physiology. July 02, 2013
    Conventional manual measurement of blood pressure (BP) in clinical practice is no longer considered to be the best method for evaluating a patient's BP status. Home BP and 24‐hour ambulatory BP monitoring are now recommended for the diagnosis and management of hypertension. Recent studies provide an alternative to conventional office BP, automated office (AO) BP, which involves multiple BP readings taken with a fully automated device with the patient resting quietly alone. AOBP is preferable to routine manual office BP in that it exhibits improved accuracy and a stronger relationship to both ambulatory BP and to target organ damage. Having the patient alone eliminates conversation between the patient and the observer, a cause of ‘white coat hypertension’. Use of an automated device improves accuracy, reduces digit preference, minimizes observer bias and facilitates the recording of multiple BP readings. Comparative BP data obtained in clinical studies in both research settings and in routine community practice support the use of a cut‐point of 135/85 mmHg for defining hypertension using AOBP, which is the same cut‐point currently recommended for awake ambulatory BP and home BP. Reduction of the white coat response using AOBP should reduce the need to monitor patients with ambulatory BP and home BP after initiation of antihypertensive therapy. There is now sufficient evidence to consider replacing manual office BP with AOBP in routine clinical practice. This article is protected by copyright. All rights reserved.
    July 02, 2013   doi: 10.1111/1440-1681.12149   open full text
  • Aldosterone, Organ Damage, and Dietary Salt.
    Cristiana Catena, GianLuca Colussi, Leonardo A. Sechi.
    Clinical and Experimental Pharmacology and Physiology. June 26, 2013
    Long‐term exposure to elevated aldosterone levels or activation of the mineralocorticoid receptors results in cardiac, vascular, and renal tissue injury with mechanisms that are independent of blood pressure levels. This evidence has been obtained in experiments conducted in hypertensive animal models and clinical studies involving patients with heart failure, essential hypertension, and primary aldosteronism. Animal studies have indicated that aldosterone causes cardiovascular and renal tissue damage only in the context of an inappropriate salt status. It has been also suggested that some of the untoward effects of high‐salt intake might depend on activation of mineralocorticoid receptors resulting from increased generation of reactive oxygen species and changes in the intracellular redox potential. Although the interaction between dietary salt intake and circulating aldosterone in causing organ damage has received robust support from the results of animal experiments, the evidence of such interaction in the clinical setting is only preliminary and will require further investigation in appropriately designed studies. This article is protected by copyright. All rights reserved.
    June 26, 2013   doi: 10.1111/1440-1681.12145   open full text
  • The clinical significance of home blood pressure measurements for the prevention and management of high blood pressure.
    Yutaka Imai, Miki Hosaka, Noha Elnagar, Michihiro Satoh.
    Clinical and Experimental Pharmacology and Physiology. June 13, 2013
    1. Ambulatory blood pressure (ABP) monitoring (M) provides BP information at many points on any particular day during unrestricted routine daily activities, whereas home blood pressure (HBP) monitoring provides a lot of BP information obtained under fixed times and conditions over a long period of time, thus mean values of HBP provide high reproducibility, and thus an overall superiority compared with ABP. 2. HBP is at least equally or better able than ABP to predict hypertensive target organ damage and prognosis of cardiovascular disease. 3. HBPM allows for ongoing disease monitoring by patients, improves adherence to antihypertensive treatment, and can provide health care providers with timely clinical data and direct and immediate feedback regarding diagnosis and treatment of hypertension. 4. HBPM provides BP information in relation to time, i.e., BP in the morning, in the evening, and at night during sleep, and it is an essential tool for the diagnosis of white‐coat and masked hypertension. 5. HBPM yields minimal alerting effects and a placebo effect, and can therefore distinguish small but significant serial changes in BP. It is thus the most practical way to monitor BP in the day‐by‐day management of hypertension. 6. The superiority of HBPM over ABPM and clinic BPM is apparent from almost all practical and clinical research perspectives. This article is protected by copyright. All rights reserved.
    June 13, 2013   doi: 10.1111/1440-1681.12142   open full text
  • Multiscale Measurement of Cardiac Energetics.
    Soyeon Goo, Toan Pham, June‐Chiew Han, Poul Nielsen, Andrew Taberner, Anthony Hickey, Denis Loiselle.
    Clinical and Experimental Pharmacology and Physiology. June 08, 2013
    We describe our laboratories’ experimental methods for interrogating cardiac energetics – at the organ (whole‐heart), tissue (trabecula) and perforated fibre (mitochondrial) levels. In whole‐heart and trabecula experiments, we focus on measuring pressure‐volume (force‐length) work and oxygen consumption (heat production) from which mechanical efficiency is derived. In both preparations, i.e., across scales differing by three orders of magnitude, we find efficiency values of 10‐15%. Mitochondrial experiments invoke a trio of titration protocols to yield information on oxygen consumption, ATP flux, membrane potential, electron leak and ROS production, the latter two of which index energy transfer inefficiencies. This article is protected by copyright. All rights reserved.
    June 08, 2013   doi: 10.1111/1440-1681.12139   open full text
  • Usefulness of ambulatory blood pressure monitoring (ABPM) in daily clinical practice: Data from the Spanish ABPM registry.
    Julian Segura, Jose R Banegas, Luis M Ruilope.
    Clinical and Experimental Pharmacology and Physiology. May 27, 2013
    1. Hypertension is one of the most important challenges for public health systems because of its high prevalence and its association with the risk of cardiovascular and renal diseases. 2. Adequate control of hypertension is low in population and medical settings, with physicians frequently misclassifying patients’ blood pressure status based on readings taken in the clinic rather than ambulatory blood pressure measurements (ABPM). 3. Data from the Spanish Society of Hypertension ABPM registry support ABPM as a feasible option in the primary care setting, providing valuable information for the diagnosis and management of hypertension. By using ABPM rather than office BP monitoring, BP control can be doubled. This is an encouraging message to clinicians, although there is still a relatively large degree of undetected controlled and uncontrolled hypertension. 4. This short review describes the design, development and main results of the Spanish Society of Hypertension ABPM registry, a project based on a large‐scale network of Spanish physicians trained in ABPM. This article is protected by copyright. All rights reserved.
    May 27, 2013   doi: 10.1111/1440-1681.12126   open full text
  • Direct Contribution of Vascular Mineralocorticoid Receptors to Blood Pressure Regulation.
    K. Barrett, A. McCurley, I.Z. Jaffe.
    Clinical and Experimental Pharmacology and Physiology. May 25, 2013
    Hypertension is an extremely prevalent cardiovascular risk factor and current antihypertensive therapies do not adequately treat hypertension in many affected individuals. Thus a better understanding of mechanisms of hypertension could lead to novel therapies. Mineralocorticoid receptors (MR) are known to regulate blood pressure by responding to aldosterone in the kidney to regulate sodium retention. Recent evidence supports a direct contribution of the vasculature to control of BP and suggests the possibility that MR antagonists may also lower blood pressure by acting on extra‐renal MR. This review summarizes existing research considering the role of vascular mineralocorticoid receptor (MR) in regulating vasoreactivity and blood pressure. Multiple studies indicate a role for vascular MR in modulating vasoconstriction and vasorelaxation. Activation of MR in vascular endothelial and smooth muscle cells leads to increased reactive oxygen species production and decreased availability of nitric oxide, important regulators of vascular reactivity. Transgenic mouse models, including an endothelial MR over‐expressing mouse and a smooth muscle cell‐specific MR knockout mouse, support a direct role for vascular MR in control of blood pressure. This new evidence demonstrating that vascular MR directly contributes to control of vasoreactivity and blood pressure supports vascular MR, and the pathways it controls, as novel therapeutic targets to treat hypertension. This article is protected by copyright. All rights reserved.
    May 25, 2013   doi: 10.1111/1440-1681.12125   open full text
  • White‐Coat Hypertension.
    Catherine A. Martin, Barry P. McGrath.
    Clinical and Experimental Pharmacology and Physiology. May 18, 2013
    Aim A number of studies have examined if WCHT is associated with increased cardiovascular risk but with definitions of WCHT that were not sufficiently robust, and results have been inconsistent. This review aims to standardise the evidence by only including studies which used a definition of WCHT consistent with international guidelines. Method Published studies were reviewed for data on vascular dysfunction, target organ damage, risk of future sustained hypertension and cardiovascular events. Findings WCHT has a population prevalence of approximately 15% and is associated with non‐smoking and slightly elevated clinic blood pressure. Compared to normotensives, subjects with WCHT are at increased cardiovascular risk due to a higher prevalence of glucose dysregulation, increased left ventricular mass index and increased risk of future diabetes and hypertension. Conclusion Management of a subject with WCHT should focus on cardiovascular risk factors, particularly glucose intolerance, not blood pressure alone. This article is protected by copyright. All rights reserved.
    May 18, 2013   doi: 10.1111/1440-1681.12114   open full text
  • Molecular insights from dysregulation of the thiazide‐sensitive WNK/SPAK/NCC pathway in the kidney: Gordon syndrome and Thiazide‐Induced Hyponatraemia.
    Mark Glover, Kevin M. O'Shaughnessy.
    Clinical and Experimental Pharmacology and Physiology. May 18, 2013
    Human blood‐pressure is dependent on balancing dietary salt intake with its excretion by the kidney. Mendelian‐syndromes of altered blood pressure demonstrate the importance of the distal nephron in this process and of the thiazide‐sensitive pathway in particular. Gordon‐Syndrome(GS), the phenotypic inverse of the salt‐wasting Gitelman syndrome, is a condition of hyperkalaemic hypertension that is reversed by low dose thiazide diuretics or a low salt diet. Variants within at least four genes (the With‐No‐lysine‐(K)‐kinases, WNKs1&4, Cullin3 and KeLch‐like3) can cause the phenotype of GS. Details are still emerging for some of these genes, but it is likely that they all cause a gain‐of‐function in the thiazide sensitive Na‐Cl Cotransporter (NCC) and hence salt retention. We discuss the key role of a kinase, STE20(sterile 20)/SPS1‐related proline/Alanine‐rich Kinase(SPAK), that functions as an intermediary between the WNK kinases and NCC and whose loss‐of‐function mutation produces a Gitelman‐type phenotype in a mouse model. In addition to Mendelian blood pressure syndromes, the study of patients who develop Thiazide‐Induced‐Hyponatraemia(TIH) may give‐further molecular insights into the role of the thiazide‐sensitive pathway for salt reabsorption. We discuss the key features of TIH including its high degree of reproducibility on rechallenge, possible genetic predisposition and mechanisms involving excessive saliuresis and water‐retention. Taken together, the study of Gordon syndrome and TIH may increase understanding of the molecular regulation of sodium trafficking via the thiazide‐sensitive pathway and have important implications for hypertensive patients, both in the identification of new anti‐hypertensive drug targets and avoidance of hyponatraemic side‐effects. This article is protected by copyright. All rights reserved.
    May 18, 2013   doi: 10.1111/1440-1681.12115   open full text
  • Extra‐renal Roles of the WNKs.
    Keith Siew, Kevin M. O'Shaughnessy.
    Clinical and Experimental Pharmacology and Physiology. May 11, 2013
    Identified over a decade ago, the With‐No‐Lysine[K] (WNK)s have been the subsequent focus of intense research into the renal handling of Na+, Cl‐, K+ and several rare monogenetic diseases. However the potential extra‐renal roles for WNKs have been less well explored. Thiazides and Gordon syndrome are known to have effects on bone mineral density, Ca2+ and PO43‐ homeostasis, which were originally assumed to be an indirect effect through the kidney. However, current data suggests a complex and direct role for WNKs in the physiology of bone. The WNKs also modulate systemic blood pressure at several levels including the vascular resistance vessels where they cause vasoconstriction by altering the abundance of the transient receptor potential canonical channel (TRPC) 3 and/or phosphorylation of the Na+‐K+‐2Cl‐ cotransporter (NKCC1) in vascular smooth muscle (VSM) cells. The WNKs and many of the cation‐coupled Cl‐ cotransporters they regulate are highly expressed in the central nervous system and recent work has suggested that WNK dysfunction may have a role in the development of autism, schizophrenia and hereditary sensory and autonomic neuropathy type 2 (HSAN2). Finally, the WNK‐Sterile 20 kinase (STE20) signalling axis represents an evolutionarily ancient mechanism for maintaining osmotic homeostasis but a rapidly expanding body of evidence also shows a role in immunity and cellular regulation. This article is protected by copyright. All rights reserved.
    May 11, 2013   doi: 10.1111/1440-1681.12108   open full text
  • Is there a role for ambulatory blood pressure monitoring in pregnancy?
    Mark Brown.
    Clinical and Experimental Pharmacology and Physiology. May 08, 2013
    Ambulatory blood pressure monitoring (ABPM) has been used in pregnancy for just over 20 years now and is generally well tolerated. Normal values have been established for different gestations; these are slightly higher than conventional BP in normal pregnancy, presumably reflecting greater activity during the 24 hr of ABPM recordings. ABPM is a better predictor than conventional BP for the development of pre‐eclampsia and fetal growth restriction but is not sensitive or specific enough to recommend ABPM for these purposes in routine practice. ABPM studies have shown that sleep hypertension is common in women with gestational hypertension or pre‐eclampsia, but detecting this in routine clinical practice does not aid usual decision making in the pregnancy, including timing of delivery. Studies utilising ABPM have found that pregnant women who are working outside the home have higher BPs than non‐working women but these data should not be interpreted to mean that not working during pregnancy will prevent gestational hypertension or pre‐eclampsia. The best role for ABPM is to determine whether women with office hypertension in early pregnancy have true (usually essential) hypertension, or white‐coat hypertension. The latter can be managed without antihypertensives and pregnancy outcomes appear good, though possibly with a slightly increased incidence of pre‐eclampsia Women who have had pre‐eclampsia are at greater life time risk for cardiovascular diseases; several years post‐partum these women have slightly higher ABPM‐measured BPs than women who had normal pregnancies and a greater propensity to metabolic syndrome. This article is protected by copyright. All rights reserved.
    May 08, 2013   doi: 10.1111/1440-1681.12106   open full text
  • Automated measurement of office, home and ambulatory blood pressure in atrial fibrillation.
    Anastasios Kollias, George S. Stergiou.
    Clinical and Experimental Pharmacology and Physiology. May 04, 2013
    Hypertension and atrial fibrillation (AF) often coexist and are strong risk factors for stroke. Current guidelines for blood pressure (BP) measurement in AF recommend repeated measurements using the auscultatory method, whereas the accuracy of the automated devices is regarded as questionable. This review presents the current evidence on the feasibility and accuracy of automated BP measurement in the presence of AF, and the potential for automated detection of undiagnosed AF during such measurements. The studies that evaluated the use of automated BP monitors in AF are limited and have significant heterogeneity in methodology and protocols. Overall, the oscillometric method is feasible for static (office or home) and ambulatory use and appears to be more accurate for systolic than diastolic BP measurement. Given that systolic hypertension is particularly common and important in the elderly, the automated BP measurement method might be acceptable for self‐home and ambulatory monitoring, but not for professional office or clinic measurement. An embedded algorithm for the detection of asymptomatic AF during routine automated BP measurement with high diagnostic accuracy has been developed and appears to be a useful screening tool for elderly hypertensives. This article is protected by copyright. All rights reserved.
    May 04, 2013   doi: 10.1111/1440-1681.12103   open full text
  • Blood pressure variability in risk stratification: what does it add?
    Kei Asayama, Rudolph Schutte, Yan Li, Tine W Hansen, Jan A Staessen.
    Clinical and Experimental Pharmacology and Physiology. April 09, 2013
    In this mini‐review, we will address the predictive value of blood pressure variability, over and beyond level of pressure, in randomly selected population samples. All reviewed studies had sufficient power, long follow‐up duration and a wide age range. We assessed blood pressure variability derived from home visit, self‐measured home pressure, and 24‐h ambulatory monitoring. The conclusions are mainly based on novel indices of blood pressure variability — variability independent of the mean, difference between maximum and minimum blood pressure, and average real variability. None of these variability indices or morning surge in blood pressure substantially refined risk profiling over and beyond the blood pressure level. In risk stratification, clinicians should concentrate on blood pressure level, the predominant risk factor modifiable by lifestyle measures and antihypertensive drug treatment. This article is protected by copyright. All rights reserved.
    April 09, 2013   doi: 10.1111/1440-1681.12091   open full text
  • Aldosterone induces p21‐regulated apoptosis via increased synthesis and secretion of tumour necrosis factor‐α in human proximal tubular cells.
    Kento Kitada, Daisuke Nakano, Hirofumi Hitomi, Hiroyuki Kobori, Kazushi Deguchi, Hirohito Mori, Tsutomu Masaki, Akira Nishiyama.
    Clinical and Experimental Pharmacology and Physiology. September 12, 2012
    Aldosterone has been shown to mediate p21‐dependent cellular senescence in rat kidney proximal tubules in vivo and in cultured human proximal tubular cells. p21‐induced senescent cells expressed higher levels of apoptotic cytokines, such as tumour necrosis factor‐α (TNF‐α), compared to non‐senescent cells. The aim of this study was to investigate the hypothesis that aldosterone increases proximal tubular apoptosis by increasing the secretion of apoptosis‐inducing factors through a p21‐dependent mechanism. Human proximal tubular cells were incubated with aldosterone (10 nmol/L) and cell senescence was detected by senescence‐associated β‐galactosidase staining and expression of p21. Apoptosis was analysed by terminal nick‐end labelling and annexin/propidium iodide staining. p21 localisation was observed by immunofluorescence. Aldosterone for 3 or 5 days increased senescence‐associated β‐galactosidase staining, expression of p21 and TNF‐α mRNAs, and secretion of TNF‐α into the culture medium. These changes were abolished by gene silencing of p21. Aldosterone failed to increase apoptotic cells at day 3, but did increase them at day 5. Neutralising antibody against TNF‐α prevented the aldosterone‐induced apoptotic changes. Aldosterone did not affect the localisation of p21. These findings indicate that aldosterone increases TNF‐α synthesis and secretion in proximal tubular cells via p21/senescence‐dependent cell phenotypic changes, and that secreted TNF‐α plays an important role as a paracrine factor in mediating cell apoptosis, indicating a possible involvement in aldosterone‐induced renal damage. © 2012 The Authors Clinical and Experimental Pharmacology and Physiology © 2012 Wiley Publishing Asia Pty Ltd
    September 12, 2012   doi: 10.1111/1440-1681.12001   open full text
  • Technetium‐99 Conjugated with Methylene Diphosphonate Inhibits RANKL‐induced Osteoclastogenesis.
    Wei Gong, Huan Dou, Xianqin Liu, Lingyun Sun, Yayi Hou.
    Clinical and Experimental Pharmacology and Physiology. August 22, 2012
    We have investigated the effects of technetium‐99 conjugated with methylene diphosphonate (99Tc‐MDP), an agent for radionuclide therapy, on receptor activator of nuclear factor‐κB ligand (RANKL)‐induced osteoclastogenesis, and explored its underlying mechanisms. A murine macrophage cell line RAW264.7 and bone marrow derived‐macrophages from C57BL/6 mice (BMMs) were used as models for osteoclastogenesis in vitro. The expression of some key factors in RANKL (50 ng/ml)‐induced osteoclastogenesis from RAW264.7 was investigated by flow cytometry and real‐time RT‐PCR. To detect multinucleated osteoclast formation, RAW264.7 was induced with RANKL for four days and BMMs were induced by RANKL (50 ng/ml) and macrophage colony‐stimulating factor (M‐CSF, 20 ng/ml) for seven days, and the cells were then stained with tartrate‐resistant acid phosphatase (TRAP). Osteoclast markers including CD51, matrix metalloproteinases 9 (MMP9) and Cathepsin K were used to evaluate osteoclastogenesis. 99Tc‐MDP (0.01 μg/ml) significantly inhibited RANKL‐induced osteoclastogenesis without any cytotoxicity. 99Tc‐MDP also dramatically abolished the appearance of multinucleated osteoclasts. Expressions of transcription factors, c‐Fos and nuclear factor of activated T‐cells (NFATc1), as well as inflammatory factors induced by RANKL were measured by real‐time RT‐PCR. 99Tc‐MDP inhibited the expressions of c‐Fos, NFATc1 and inflammatory factors such as interleukin 6 (IL‐6), tumor necrosis factor α (TNF‐α) and interleukin 1β (IL‐1β). Moreover, 99Tc‐MDP also inhibited the activation of mitogen‐activated protein kinases (MAPKs) in RAW264.7 cells under RANKL stimulation. In conclusion, 99Tc‐MDP possesses anti‐osteoclastogenic activity on RANKL‐induced osteoclast formation. © 2012 The Authors Clinical and Experimental Pharmacology and Physiology © 2012 Blackwell Publishing Asia Pty Ltd
    August 22, 2012   doi: 10.1111/1440-1681.12006   open full text
  • Matrix metalloproteinase‐2 inhibition by temocapril and its important role in peritoneal transport.
    Daisuke Yamamoto, Shinji Takai, Tetsu Akimoto, Ichiro Hirahara, Chiharu Ito, Shigeaki Muto, Eiji Kusano.
    Clinical and Experimental Pharmacology and Physiology. August 18, 2012
    (MMP)‐2 plays an important role in tissue remodeling during peritoneal injury caused by peritoneal dialysis (PD) therapy, but MMP‐2 inhibitors have not yet been used clinically. Recently, it was reported that captopril, an angiotensin converting enzyme (ACE) inhibitor, could inhibit MMP‐2 To investigate the potential usefulness of ACE inhibitors during PD therapy, molecular interaction between the MMP‐2 active site and the active form of temocapril (temocaprilat) was studied using molecular modeling. Furthermore, the effect of temocapril on MMP‐2 activity in peritoneal effluents and the peritoneal solute transport rate of PD patients were measured Temocaprilat bound to the MMP‐2 active centre and recognized two hydrophobic substrate‐binding sites in the MMP‐2 molecular model. MMP‐2 activity in peritoneal effluents was directly inhibited by temocaprilat (IC50; 0.47 μmol/L). In one patient given temocapril, the peritoneal solute transport rate decreased gradually during PD may prove to be an important candidate for development as a novel therapeutic agent for MMP‐2 inhibition, to prevent peritoneal injury caused by PD © 2012 The Authors Clinical and Experimental Pharmacology and Physiology © 2012 Blackwell Publishing Asia Pty Ltd
    August 18, 2012   doi: 10.1111/1440-1681.12003   open full text
  • Adenosine A2A receptor antagonists are broad facilitators of antinicotinic neuromuscular blockade monitored either with 2‐Hz train‐of‐four or 50‐Hz tetanic stimuli.
    MW Pereira, P Correia‐de‐Sá, W Alves‐Do‐Prado.
    Clinical and Experimental Pharmacology and Physiology. August 18, 2012
    he 2‐Hz train‐of‐four ratio (TOFratio) is used to monitor the degree of patient curarization. Using a rat phrenic nerve‐hemidiaphragm preparation, we showed that antinicotinic agents, such as hexamethonium, D‐tubocurarine, and pancuronium but not cisatracurium, decreased contractions produced by physiological nerve activity patterns (50 Hz) more efficiently than those caused by 2‐Hz trains. Uncertainty about the usefulness of the TOFratio to control safe recovery from curarization prompted us to investigate the muscarinic and adenosine neuromodulation of tetanic (50 Hz) fade induced by antinicotinic agents at concentrations that cause a 25% reduction in the TOFratio (TOFfade). Tetanic fade caused by D‐tubocurarine, pancuronium, and hexamethonium was attenuated by blocking presynaptic inhibitory muscarinic M2 and adenosine A1 receptors with methoctramine and DPCPX, respectively. These compounds enhanced rather than decreased tetanic fade induced by cisatracurium, but they consistently attenuated cisatracurium‐induced TOFfade. The effect of the M1 antagonist pirenzepine on fade produced by antinicotinic agents at 50‐Hz was opposite to that observed with TOF stimulation. Blockade of adenosine A2A receptors with ZM 241385 attenuated TOFfade caused by all tested antinicotinic drugs, with the exception of the “pure” presynaptic nicotinic antagonist hexamethonium. ZM 241385 was the only compound tested in this series that facilitated recovery from tetanic fade produced by cisatracurium. The data suggest that distinct antinicotinic relaxants interfere with fine‐tuning neuromuscular adaptations to motor nerve stimulation patterns via presynaptic muscarinic and adenosine receptor activation. These results support the use of A2A receptor antagonists together with atropine to facilitate recovery from antinicotinic neuromuscular blockade. © 2012 The Authors Clinical and Experimental Pharmacology and Physiology © 2012 Blackwell Publishing Asia Pty Ltd
    August 18, 2012   doi: 10.1111/1440-1681.12004   open full text
  • Inhibition of extracellular signal‐regulated kinase activity improves cognitive function in Tg2576 mice.
    Peng Jin, Dong‐Young Choi, Jin Tae Hong.
    Clinical and Experimental Pharmacology and Physiology. August 06, 2012
    Deposition of β‐amyloid (Aβ) peptide is a defining pathological hallmark of Alzheimer's disease (AD) and is involved in memory impairment. Evidence suggests that activation of an extracellular signal‐regulated kinase (ERK) pathway is related to Aβ accumulation. Thus, the aim of the present study was to investigate the effects of an ERK inhibitor (U0126) on amyloidogenesis and cognitive function in Tg2576 mice. Tg2576 mice received injection of U0126 (20 mg/kg, i.p.) or vehicle (1% dimethyl sulfoxide in sterile saline) once a day for 7 days and then cognitive function was assessed by the Morris water maze test and passive avoidance test. In addition, immunostainings, western blots, Enzyme‐linked immunosorbent assay (ELISA) and enzyme activity assay were employed to examine a degree of Aβ deposition in the brains. Our results showed that U0126 attenuated memory impairment and inhibited Aβ deposition in the brains of Tg2576 mice. Further experiments revealed that the inhibition of Aβ deposition by U0126 was due to a reduction in β‐secretase and amyloid precursor protein expression in the brains of U0126‐treated Tg2576 mice. These results suggest that the ERK pathway is associated with Aβ accumulation and consequent memory dysfunction in Tg2576 mice and that inhibition of the ERK pathway may be an appropriate intervention in the treatment of AD © 2012 The Authors Clinical and Experimental Pharmacology and Physiology © 2012 Blackwell Publishing Asia Pty Ltd
    August 06, 2012   doi: 10.1111/cep.12000   open full text
  • Chronic exercise partially restores the transmural heterogeneity of action potential duration in left ventricular myocytes of spontaneous hypertensive rats.
    Danilo Roman‐Campos, Miguel A. Carneiro‐Júnior, Thales N. Prímola‐Gomes, Karina A. Silva, Judson F. Quintão‐Júnior, Antonio N.S. Gondim, Hugo L. Duarte, Jader S. Cruz, Antonio J. Natali.
    Clinical and Experimental Pharmacology and Physiology. January 03, 2012
    1. Hypertension leads to electrophysiological changes in the heart. Chronic exercise induced by a treadmill‐running program (TRP) is considered as a potential non‐pharmacological treatment for hypertension and may have implications in heart remodeling. However, it is not known if TRP is able to improve the electrophysiological properties of the heart in spontaneously hypertensive rats (SHR). In the present study, we tested whether TRP affects the electrical properties of left ventricular (LV) myocytes isolated from different layers of the LV wall of SHR.2. Male SHR were divided into exercised (chronic treadmill running for 8 weeks; CEX‐SHR) and sedentary (SED‐SHR) groups. Age matched normotensive Wistar male rats served as controls. Action potentials (AP) and transient outward potassium current (Ito) were recorded in sub‐epicardial (EPI) and sub‐endocardial (ENDO) LV myocytes.3. We found that in normotensive controls, AP duration (APD) in ENDO cells were longer than in EPI cells. Such a transmural heterogeneity in LV was not observed in sedentary SHR but was partially restored in SHR subject to chronic exercise. This partial recovery was associated with an increase in Ito density in EPI cells but not in ENDO cells. The electrophysiological changes observed in the CEX‐SHR group were not accompanied by amelioration of systolic blood pressure neither by reduction in heart hypertrophy.4. These findings imply that TRP is able to improve electrophysiological parameters of isolated cardiac myocytes in SHR. Such adaptation contributes to an overall improvement of heart physiology in this model.© 2012 The Authors Clinical and Experimental Pharmacology and Physiology© 2012 Blackwell Publishing Asia Pty Ltd
    January 03, 2012   doi: 10.1111/j.1440-1681.2012.05669.x   open full text