Resveratrol attenuates hydrogen peroxide-induced injury of rat ovarian 1 granulosa-lutein cells by resisting oxidative stress via the SIRT1/Nrf2/ARE 2 signaling pathway

Abstract Background : The ovarian granulosa-lutein cells injury and apoptosis induced by oxidative stress may be responsible for female luteal phase deficiency. The antioxidant function of resveratrol has been confirmed, however, its effect on the expression of antioxidant enzymes and regulatory mechanism in ovarian granulosa-lutein cells remains unclear. Results : The H2O2 treatment induced the primary ovarian granulosa-lutein cells injury shown as decreased cell viability, impaired cellular morphology, decreased levels of progesterone and estradiol. The H2O2 treatment also exacerbated cell apoptosis demonstrated as more apoptotic cells stained by Hoechst staining, decreased level of anti-apoptosis protein Bcl-2 and increased level of pro-apoptosis protein Bax. These effects of cell injury and apoptosis induced by H2O2 can be ameliorated by resveratrol. Resveratrol also alleviated oxidative stress induced by H2O2 supported by decreased superoxide anion and cellular total ROS, decreased levels of malondialdehyde and protein carbonyl, increased total antioxidant capacity and biological activity of SOD. Western blot results demonstrated resveratrol reversed the H2O2-induced decrease in levels of antioxidant enzymes containing ARE sequences and activated SIRT1/Nrf2 pathway. Further treatment by siRNA-Nrf2 suggested resveratrol was unable to activate the expression of antioxidant enzymes under a condition of inhibition of Nrf2. Conclusions : In conclusion, these results indicated that resveratrol attenuated oxidative stress to protect H2O2-induced rat ovarian granulosa-lutein cells injury and apoptosis via SIRT1/Nrf2/ARE signaling pathway.