Roles of SIRT1 in granulosa cell apoptosis during the process of follicular atresia in porcine ovary.

Ovarian follicular atresia is characterized by granulosa cell apoptosis, however, the exact mechanism is still unclear. Sirtuin 1 (SIRT1) is a NAD(+)-dependent deacetylase, which is associated with apoptosis in several of cell types, but its exact role in ovarian granulosa cell apoptosis is not clearly defined. In present study, we identified the involvement of SIRT1 in the process of follicle degeneration, which is known as "follicular atresia", both from in vivo models and cell culture data. The results of immunohistochemistry showed that SIRT1 was widely detected in non-apoptotic granulosa cells of follicles, but significantly decreased during the process of granulosa cell apoptosis. Quantitative real-time PCR and Western blot analysis showed that the expression levels of SIRT1 mRNA and protein were increased (P<0.05) during follicular atresia. In order to provide more evidences elucidating the roles of SIRT1 during the process of follicular atresia, granulosa cells were cultured in vitro with resveratrol which acts as a potent activator of SIRT1. Results showed that resveratrol caused a dose-dependent increase in both SIRT1 mRNA and protein levels. Meanwhile, apoptotic rate of granulosa cell was increased (P<0.01) in a dose-dependent manner. Additionally, resveratrol significantly increased the expression levels of Caspase-3 (P<0.01) and Bax mRNA (P<0.01), while Bcl-2 mRNA level was significantly decreased (P<0.01). Thus, our results suggest that SIRT1 may play important roles in the regulation of granulosa cell apoptosis during follicular atresia in porcine ovary.