不同浓度白藜芦醇激动SIRT1对卵巢颗粒细胞凋亡的调控

Effects of different concentrations of resveratrol on SIRT1 activation in ovarian granulosa cells

目的:研究SIRT1激动剂白藜芦醇(RES)调控卵巢颗粒细胞凋亡的最佳作用浓度。方法:选取人卵巢颗粒细胞瘤细胞系COV434细胞,分别采用不同浓度20、50、75μmol/L和100μmol/L RES激动SIRT1,qRT-PCR法检测RES对SIRT1基因的激活作用,Western blot法检测凋亡相关蛋白Caspase-3及Bcl-2表达,CCK-8毒性实验检测细胞增殖状况,Annxin V-FITC/PI双染流式细胞术测定细胞凋亡。结果:随着RES作用浓度的增加,COV434细胞数增多,SIRT1表达增高,细胞凋亡抑制因子Bcl-2表达增加,促凋亡因子Caspase-3表达降低,细胞凋亡率降低,细胞增殖率升高;RES作用浓度达50μmol/L时,与对照组比较,差异有统计学意义(P<0.05)。结论:50μmol/L RES作为研究SIRT1对卵巢颗粒细胞凋亡作用的有效激活浓度,可作为研究早发性卵巢功能不全的药物干预浓度依据。

Objective： To study the effect of resveratrol on the apoptosis of ovarian granulosa cells by agitating SIRT1.Methods： Human ovarian granulosa cell tumor COV434 cell line were treated with different concentrations of 20μmol/L,50μmol/L,75μmol/L,100μmol/L resveratrol to stimulate SIRT1.qRT-PCR was used to evaluate the activation of SIRT1 gene by resveratrol.The expressions of Caspase-3 and Bcl-2 were detected by Western blot.The cell proliferation and apoptosis were detected by CCK-8 toxicity test and flow cytometry.Results： In the COV434 cells,the expression of SIRT1 was increased with the increase of the concentration of resveratrol,and the expression of Bcl-2 was increased and Caspase-3 was decreased. Flow cytometry showed that apoptosis decreased with rising drug concentration. The result of CCK-8 showed that the cell proliferation was in a concentration-time-dependent manner.The expression of SIRT1 mRNA was significantly increased when the concentration of resveratrol reached 50μmol/L,the apoptotic rate of treating with resveratrol was decreased compared with the blank control group and with the increase of concentration,the apoptosis rate decreased gradually.Conclusion： The effective concentration of 50μmol/L as an effective activator for the study of SIRT1 on ovarian granulosa cells is used to be a new drug for intervention of premature ovarian insufficiency.