雄激素诱导小鼠颗粒细胞凋亡及其机制探讨

Effect and mechanism of androgen on cultured mouse ovarian granulosa cell apoptosis

目的通过睾酮诱导小鼠颗粒细胞凋亡,观察热休克蛋白(HSP)10在高雄激素下的表达,探讨HSP10在高雄激素诱导颗粒细胞凋亡中的作用。方法原代培养小鼠颗粒细胞,用0、10-8M、10-6M、10-5M睾酮处理24h,检测HSP10基因表达;处理48h,检测细胞活力及活性Caspase-3的表达。用AdCMV-H1-siRNA/HSP10腺病毒在小鼠颗粒细胞中干涉HSP10的表达,MTT检测细胞活力,流式细胞术检测细胞周期,并检测Ki67、Bcl-2、Bax基因水平变化。结果与0M睾酮处理组比较,10-6M、10-5M睾酮处理组颗粒细胞的细胞活力降低,且Caspase-3表达增加,HSP10表达降低(P<0.05);感染AdCMV-H1-siRNA/HSP10腺病毒后,颗粒细胞的细胞活力降低,且细胞增殖阻滞在G2期,Bcl-2、Ki67表达降低而Bax表达增加(P<0.05)。结论高雄激素可诱导小鼠颗粒细胞凋亡。其可能的机制是睾酮能降低HSP10表达,经由降低Bcl-2和增加Bax表达的通路。

Objective To investigate the roles of heat shock protein 10（HSP10） in apoptosis of granulosa cells induced by androgen.Methods After the mouse primary cultured granulosa cells were exposed to 0,10-8M,10-6M and 10-5M testosterone for 24 h,the expression of HSP10 gene was detected.The cell viability and expression of active Caspase-3 were examined after 48 h exposure,which was infected by AdCMV-H1-siRNA/HSP10 adenovirus to downregulate the expression of HSP10,and cell viability was detected by MTT,cell cycle was detected by flow cytometry,and gene expressions of Ki67,Bcl-2 and Bax were detected by RT-PCR.Results Compared with group 0 M testosterone,the cell viability reduced,Caspase-3 increased and HSP10 expression decreased in the testosterone-treated groups of 10-6M and 10-5M（P0.05）.After infected with AdCMV-H1-siRNA/HSP10,the cell viability reduced,cell cycle was arrested at G2 with decreased expressions of Bcl-2 and Ki67 genes and increased expression of Bax gene（P0.05）.Conclusion Hyperandrogen can induce the apoptosis of mouse granulosa cells,which may be related to reduced HSP10 expression by reducing Bcl-2 expression and increasing Bax expression.