Abstract:Objective: To investigate the effect of plant extract resveratrol on autophagy of the granulosa cells in rat model with polycystic ovary syndrome and to explore its regulation on SIRT1/AMPK pathway. Methods: Ovarian granulosa cells of Sprague Dawley rats with polycystic ovary syndrome (PCOS) were cultured in vitro and treated with different concentrations of resveratrol (5, 10, 20, 40 μmol/L) for different times (24, 48, 72 h). The proliferation rate of ovarian granulosa cells was observed by MTT method, the changes of intracellular autophagy were observed by electron microscope, and the expression changes of silent information regulator T1 (SIRT1), adenosine monophosphate-activated protein kinase (AMPK), phosphorylated AMPK (pAMPK), Beclin1 and p62 were detected by Western blot. Finally, the expression of the above proteins in the cells was detected after the application of EX527, as a specific SIRT1 inhibitor. Results: MTT results showed that resveratrol promoted the proliferation rates of the granulosa cells of PCOS rats (P<0.05), and it was in a concentration-time dependent manner (P<0.05). Electron microscope showed that resveratrol could reduce the number of autophagosome in cells (P<0.05). Western blot results showed that resveratrol could enhance the protein expression levels of SIRT1 and pAMPK in ovarian granulosa cells (P<0.05), inhibit the expression of Beclin1 (P<0.05), and increase the expression of p62. After combined with SIRT1 specific inhibitor EX527, the protein expression levels of SIRT1 and pAMPK in granulosa cells decreased significantly, the expression of Beclin1 increased significantly (P<0.05), while p62 decreased significantly (P<0.05). Conclusion: Resveratrol could inhibit the level of autophagy and promote the proliferation of ovarian granulosa cells of the rats with PCOS, which is closely related to the activation of SIRT1/AMPK pathway by resveratrol.
