【摘要】 目的：探讨睾酮在体外对颗粒细胞凋亡的诱导作用，并阐明其作用机制。方法：噻唑蓝（MTT）比色法检测人卵巢颗粒细胞SVOG的细胞增殖抑制率，筛选最佳药物作用浓度后，将SVOG细胞分为对照组、睾酮组(1.0×10^-5 mol·L^-1睾酮)、睾酮+氟他胺组(1.0×10^-5 mol·L^-1睾酮+1.0×10^-5 mol·L^-1氟他胺)、睾酮+牛磺熊去氧胆酸（TUDCA）组(1.0×10^-5 mol·L^-1睾酮+1.5 g·L^-1 TUDCA)。采用实时荧光定量PCR （RT-qPCR）法检测各组SVOG细胞中剪切型X盒结合蛋白1（XBP1s）、激活转录因子4 （ATF4）、转录因子C/EBP同源蛋白（CHOP）和死亡受体5（DR5）mRNA表达水平，流式细胞术检测各组SVOG细胞凋亡率。结果：与对照组比较，睾酮组SVOG细胞形态改变，异型细胞增多，胞质中出现较多颗粒，培养基中细胞碎片增多，SVOG细胞增殖抑制率升高（P<0.05）；与睾酮组比较，睾酮+氟他胺组和睾酮+TUDCA组SVOG细胞形态变化减少，细胞增殖抑制率降低（P<0.05）；流式细胞术，与对照组比较，睾酮组SVOG细胞凋亡率升高（P<0.05）；与睾酮组比较，睾酮+氟他胺组和睾酮+TUDCA组SVOG细胞凋亡率降低（P<0.05）。RT-qPCR法，作用48 h后，与对照组比较，睾酮组SVOG细胞中XBP1s、ATF4、CHOP和DR5 mRNA表达水平均明显升高（P<0.05）；与睾酮组比较，睾酮+氟他胺组和睾酮+TUDCA组SVOG细胞中XBP1s、ATF4、CHOP和DR5 mRNA表达水平降低（P<0.05）。结论：睾酮可诱导人卵巢颗粒细胞凋亡并引起卵泡闭锁，其作用机制可能是雄激素与雄激素受体（AR）结合激活CHOP-DR5通路，诱导内质网应激（ERS）发生，促进颗粒细胞凋亡。更多还原

【Abstract】 Objective:To discuss the inductive effect of testosterone on the apoptosis of the granulosa cells in vitro,and to clarify its mechanism. Methods:The inhibitory rates of proliferation of human ovarian granulosa cells SVOG were detected by methyl thiazolyl tetrazolium（MTT） assay. After screening the optimal concentrations of the drugs, the SVOG cells were divided into control group, testosterone group(1. 0×10^-5 mol·L^-1 testosterone),testosterone+fluorothiamine group(1. 0×10^-5 mol·L^-1 testosterone+1. 0×10^-5 mol·L^-1 fluorothiamine),and testosterone+tauroursodeoxycholic acid（TUDCA）group(1. 0×10^-5 mol·L^-1 testosterone+1. 5 g·L^-1 TUDCA). Real-time fluorescence quantitative PCR（RT-qPCR） method was used to detect the expression levels of spliced X-box binding protein 1（XBP1s）, activating transcription factor 4（ATF4）, transcription factor C/EBP homologous protein（CHOP）, and death receptor 5（DR5） mRNA in the SVOG cells in various groups; flow cytometry was used to detect the apoptotic rates of the SVOG cells in various groups. Results:Compared with control group, the SVOG cells in testosterone group showed morphological changes, the number of atypical cells was increased, the granularity in the cytoplasm was increased, the cell fragments in the culture medium was increased, and the inhibitory rate of proliferation of the SVOG cells was increased（P<0. 05）. Compared with testosterone group, the morphological changes of the cells in testosterone +fluorothiamine group and testosterone+TUDCA group were decreased and inhibitory rates of proliferation of the cells were decreased（P<0. 05）.The flow cytometry results showed that compared with control group, the apoptotic rate of the SVOG cells in testosterone group was significantly increased（P<0. 05）. Compared with testosterone group, the apoptotic rates of the SVOG cells in testosterone + TUDCA group and testosterone + fluorothiamine group were significantly decreased（P<0. 05）. The RT-qPCR analysis results showed that after treated for 48 h, compared with control group, the expression levels of XBP1s, ATF4, CHOP, and DR5 mRNA in the SVOG cells in testosterone group were increased（P<0. 05）. Compared with testosterone group, the expression levels of XBP1s,ATF4,CHOP, and DR5 mRNA in the SVOG cells in testosterone+TUDCA group and testosterone+fluorothiamine group were decreased（P<0. 05）. Conclusion:Testosterone can induce the apoptosis of human ovarian granulosa cells and cause follicular atresia,and its mechanism may be that androgen combined with adrogen receptor（AR） activates the CHOP-DR5 pathway, induces endoplasmic reticulum stress（ERS）, and promotes the apoptosis of the granulosa cells.更多还原