--- - |2- The selective and potent inhibitor of Sirt1, ex‐527, reduces the infarction volume of ischemic brains and improves the survival rate. Treatment with ex‐527 effectively abolishes the elevation of the critical regulators of necroptosis. Ex‐527 relieves ischemia‐induced perturbation of necroptosis‐associated metabolic enzyme activity downstream. Necroptosis, a novel type of programmed cell death, is involved in ischemia–reperfusion‐induced brain injury. Sirtuin 1 (Sirt1), as a well‐known member of histone deacetylase class III, plays pivotal roles in inflammation, metabolism, and neuron loss in cerebral ischemia. We explored the relationship between Sirt1 and the necroptosis signaling pathway and its downstream events by administration of ex‐527, as a selective and potent inhibitor of Sirt1, and necrostatin‐1 (nec‐1), as a necroptosis inhibitor, in an animal model of focal cerebral ischemia. Our data showed different patterns of sirt1 and necroptosis critical regulators, including receptor‐interacting protein kinase 3 and mixed lineage kinase domain–like protein gene expressions in the prefrontal cortex and the hippocampus after ischemia–reperfusion. We found that ex‐527 microinjection reduces the infarction volume of ischemic brains and improves the survival rate, but not stroke‐associated neurological deficits. Additionally, treatment with ex‐527 effectively abolished the elevation of the critical regulators of necroptosis, whereas necroptosis inhibition through nec‐1 microinjection did not influence Sirt1 expression levels. Our data also demonstrated that the ex‐527 relieves ischemia‐induced perturbation of necroptosis‐associated metabolic enzymes activity in downstream. This study provides a new approach to the possible neuroprotective potential of ex‐527 orchestrated by necroptosis pathway inhibition to alleviate ischemia–reperfusion brain injury. - 'Journal of Cellular Physiology, Volume 234, Issue 2, Page 1816-1826, February 2019. '