Effect of resveratrol on oxidative stress of rat primary cortical neurons during different time windows of oxygen-glucose deprivation/reperfusion injury

Abstract

Objective To investigate the effect of resveratrol on oxidative stress of rat primary cortical neurons during different time windows of oxygen-glucose deprivation/reperfusion(OGD/RP) injury.MethodsRat cortical neurons were cultured under oxygen and glucose deprivation for 150 min and returned to normal culture for 24 h.The experiment was divided into 6 groups,including the normal group,model group,pre-treatment group(treated with resveratrol for 24 h prior to OGD),OGD-treatment group(treated with resveratrol during 150 min of OGD and 24 h of reperfusion),post-treatment group(treated with resveratrol during 24 h of reperfusion),and the whole processing group(treated with resveratrol for 24 h prior to OGD,during 150 min of OGD,and 24 h of reperfusion).Invert microscope was used to observe cell morphology.Chemical colorimetry was used to detect the activity of superoxide dismutase(SOD) and the content of nitric oxide(NO).Immunofluorescence was used to detect the nuclear translocation of nuclear factor erythroid 2-related factor 2(Nrf2).Western blotting analysis was used reveal the the protein expressions of Nrf2,heme oxygenase-1(HO-1) and NADP(H):quinone oxidoreductase-1(NQO-1).ResultsThe stereoscopic effect and refraction of neurons were enhanced at the sixth day of culture.Compared with the model group,resveratrol of various concentrations(10,20,40,60 and 80 μmol/L) significantly elevated the activity of SOD and decreased NO content in the whole processing group(P 0.05),with the effect being the best when at 40 μmol/L.Compared with model group,resveratrol significantly alleviated neuronal OGD/RP injury,promoted translocation of Nrf2 from the cytoplasm into the nuclei,and upregulated Nrf2,HO-1 and NQO-1 protein expression at all stages of OGD/RP injury,with the best effect found in the whole processing group,followed by the pre-treatment group.ConclusionResveratrol has a dose-dependent anti-oxidative stress effect on rat cortical neurons during OGD/RP injury,with the best effect seen in the whole processing group,followed by the pre-treatment group.The mechanism might be associated with activation of Nrf2/antioxidant response element(ARE) signaling pathway and the sbusequent upregulation of antioxidant protein expression.