Abstract:Objective: To investigate the effect of the plant extract resveratrol on autophagy of granulosa cells in the rats with polycystic ovarian syndrome (PCOS) and to explore its mechanism of action. Methods: Forty female SD (Sprague Dawley) rats were randomized into two groups: PCOS model group (n=30) and model control group (n=10). PCOS granulosa cells identified and confirmed by immunocytochemistry were cultured in vitro, and dealt with different concentrations of resveratrol, and the changes of cell proliferation rate were observed by CCK-8. Electron microscope was used for the changes of autophagy level in granular cells. Western blot was used to detect the expression of light chain 3Ⅰ (LC3Ⅰ), light chain 3Ⅱ (LC3Ⅱ), Beclin1, total (phosphorylated) protein kinase B[T (p) -Akt], and total (phosphorylated) mammalian target of rapamyoin [T (p) -mTOR], and the expression of these proteins in cells after the action of Akt inhibitor LY294002 was detected. Results: Compared with the model control group, the immunocytochemical results showed that the expression of follicle-stimulating hormone (FSH) in ovarian granulosa cells in the model group was significantly enhanced, which indicated that the model was successfully constructed and isolated. The CCK-8 results showed that resveratrol could improve the proliferation rate of granulosa cells, with concentration-time dependence. The Western blot results showed that resveratrol could decrease the expression of LC3Ⅰ, LC3Ⅱand Beclin1, but increase the protein expression of p-Akt and p-mTOR. After LY249002 treatment, the expression of LC3I, LC3Ⅱand Beclin1 increased significantly, while the expression of p-Akt and p-mTOR decreased significantly. Conclusion: Resveratrol can significantly reduce the autophagy level of ovarian granulosa cells in PCOS and affect the cell proliferation rate, and its mechanism may be closely related to the upregulation of Akt/mTOR.
