高雄激素对性成熟前小鼠颗粒细胞增殖和凋亡的影响  

Effect of hyperandrogen on the proliferation and apoptosis of ovarian granulosa cells of pre-adolescent mouse

作　　者：江帆[1] 李梅[1] 赵考考[1] 刁飞扬[1] 马翔[1] 崔毓桂[1] 刘嘉茵[1] 

机构地区：[1]南京医科大学第一附属医院生殖医学科,210029

出　　处：《中华临床医师杂志（电子版）》2012年第5期73-77,共5页Chinese Journal of Clinicians(Electronic Edition)

基　　金：国家自然科学基金(81070465);973项目(2012CB944902)

摘　　要：目的观察高浓度雄激素对性成熟前小鼠卵巢颗粒细胞增殖、凋亡的影响,初步探讨其机制,为研究多囊卵巢综合征(PCOS)病理生理机制积累更多的实验资料。方法体外培养21 d龄小鼠卵巢颗粒细胞,用不同浓度的睾酮(10-5,10-6,10-8mol/L)作用24、48、72 h,噻唑蓝(MTT)比色法检测细胞活力,流式细胞术检测细胞周期和细胞凋亡,Realtime PCR检测Caspase-3/9、Bcl-2、Bax等凋亡因子的变化。结果 10-5mol/L睾酮显著增加颗粒细胞的细胞活力;10-5mol/L睾酮作用48 h,细胞周期由G1期进入S期,细胞早期凋亡率减少1.36%,Caspase-3活性降低,Caspase-3/9、Bcl-2、Bax mRNA水平表达均下降。结论高浓度雄激素增加性成熟前的卵巢颗粒细胞的细胞活力,促进细胞增殖,抑制细胞凋亡,其作用可能通过抑制线粒体通路相关的凋亡因子的表达。本研究提示,性成熟前的高雄激素作用,可能是导致青春期PCOS发生的原因之一。Objective To observe the effect of testosterone on the proliferation and apoptosis of ovarian granulose cells（GCs）of pre-adolescent mouse,so as to investigate the pathophysiological mechanism of adolescent polycystic ovary syndrome（PCOS）.Methods GCs from mouse pre-adolescent ovary were differently treated with testosterone at higher concentration of 10-5,10-6,10-8 mol/L for 24,48,72 h.Cell viability and growth rate of GCs were assayed by MTT methods.After exposure to testosterone（10-5 mol/L）for 48 h,cell cycle and apoptosis rate of GCs were detected by flow cytometry.Caspase-3 activity was detected by absorption spectrometry.Expressions of Caspase-3/9,Bcl-2 and Bax mRNAs in the treated GCs were determined by real-time RT-RCR.Results Cell viability of pre-adolescent mouse GCs was significantly increased in the group treated with testosterone at 10-5 mol/L（P〈0.05）.When compared with the control group,the cell cycle from G1 phase into S phase in the testosterone exposure group significantly changed,while the early apoptosis rate significantly decreased by 1.36%（P〈0.05）.Caspase-3 activity significantly decreased in the testosterone treated group,and the expression levels of Caspase-3/9,Bcl-2 and Bax mRNAs significantly decreased（P〈0.05）.Conclusions Androgen at higher level can promote cell viability and inhibit the early apoptosis of GCs by inhibiting the mitochondrial pathway of apoptosis-related factor expression.This study suggests that hyperandrogenism in pre-adolescent ovary may be part of pathophysiological mechanism of adolescent PCOS.

关 键 词：多囊卵巢综合征 粒层细胞 细胞增殖 细胞凋亡 雄激素增多症 

分 类 号：R711.75[医药卫生—妇产科学]