Spexin is a central modulator of nociception. The aim of the present study was to investigate the effect of intra hippocampal CA3 (IHCA3) injection of spexin and spexin‐progesterone co‐administration on pain sensitivity in ovariectomized rat. Thirty‐five adult female rats were divided into five groups. Sham: the animals received injection of 0.5 μL ACSF by IHCA3. Experiments 1 and 2: the animals received injection of 0.5 μL of spexin bilaterally (10 and 30 nM respectively). Experiments 3 and 4: the animals received injection of 0.5 μL of spexin bilaterally (10 and 30 nM respectively) + S.C. injection of progesterone (5 mg/kg). Ovariectomy was performed in all groups to eliminate the effects of cyclic changes in the female rats. The formalin test (formalin 2.5%) was performed following the administration of spexin and progesterone. Results showed that bilateral injection of spexin in IHCA3 at both concentrations a significant (P<0.05) decrease in the pain sensitivity in the two phases of formalin test. Similarly, the bilateral injection of spexin in IHCA3 at both concentrations following the S.C. injection of progesterone significantly (P<0.05) decreases pain sensitivity in two phases of the formalin test. This pain attenuation due to the co‐administration of spexin and progesterone was more potent than spexin‐induced analgesia. According to the present results, spexin has a modulatory effect on pain sensitivity, which becomes more pronounced by progesterone administration. This article is protected by copyright. All rights reserved.