孕酮回植时间对小鼠月经样模型中子宫内膜基质细胞凋亡的影响

摘要

目的:探讨在小鼠月经样模型中孕酮回植对子宫内膜基质细胞凋亡的影响.方法:在月经模型小鼠月经发生关键时期的前后,即孕酮撤退后的12h和16h回植孕酮,24h收集小鼠子宫角,TUNEL法原位检测小鼠子宫内膜基质细胞的凋亡；Western Blotting法检测凋亡相关蛋白B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)和半胱氨酸蛋白酶-3(Caspase-3)的表达.结果:孕酮撤退后12h回植孕酮,小鼠子宫内膜中发生凋亡的基质细胞数远低于16h回植组和对照组；而16h回植孕酮则子宫内膜基质细胞的凋亡仅仅稍低于对照组.凋亡抑制蛋白Bcl-2的表达,12h回植组明显高于16h回植组和对照组(P＜0.01),而后两组之间比较差异无统计学意义(P＞0.05).凋亡促进蛋白Bax和Caspase-3的表达,12h回植组均低于16h回植组和对照组(P均＜0.01),而后两组之间比较差异无统计学意义(P均＞0.05).结论:在月经发生关键时期之前,即孕酮撤退后12h回植孕酮可能通过上调Bcl-2/Bax比例而阻断Caspase-3的蛋白表达,从而有效抑制小鼠子宫内膜基质细胞的凋亡,阻断月经发生的进程,而16h回植则不能阻断小鼠月经的发生.%Objective:To explore effects of progesterone replacement in different time after progesterone withdrawal on endometrial stromal cells apoptosis in mice menstrual-like model.Methods:Based on determined of the critical period of progesterone withdrawal precedes endometrial breakdown and shedding in mouse menstrual-like model,progesterone added back to mouse before (at 12h) or after (at 16h) the critical period(progesterone withdrawal).Mouse uterine horns were collected for the detection of mouse endometrial stromal cell apoptosis in situ by TUNEL technology.Moreover,expressions of apoptoticrelated proteins,Bcl-2,Bax and Caspase-3,were evaluated by Western Blotting in mouse endometrial stromal cells.Results:When progesterone added back to the mouse at 12h after progesterone withdrawal,the number of apoptotic cells was significantly reduced in mouse endometrial stromal cells but that was not decreased obviously when progesterone was replaced at 16h.The expression of Bcl-2,a antiapoptotic protein,in 12h replacement group was significantly higher than those of 16h replacement group and the control group (P ＜ 0.01).While there was no significant difference between the latter two groups (P ＞ 0.05).As for proapoptotic protein,expressions of Bax and Caspase-3 in 12h replacement group were significantly lower than those in 16h replacement group and the control group (P all ＞0.05),while no differences were found between the latter two groups(P all ＜ 0.01).Conclusion:Before the critical period of precedes endometrial breakdown and shedding,i.e.,progesterone replacement at 12h after progesterone withdrawal,can inhibit the apoptosis of endometrial stromal cells by raising the Bcl-2/Bax proportion and blocking the expression of Caspase-3,resulting in blocking the process of menstrual like changes in mouse.