miR-34a调节人卵巢颗粒细胞凋亡

发布时间：2018-01-09 20:23

本文关键词：miR-34a调节人卵巢颗粒细胞凋亡　出处：《江苏大学学报(医学版)》2016年06期 　论文类型：期刊论文

【摘要】：目的:探讨miR-34a对人卵巢颗粒细胞的作用及其机制。方法:体外培养人卵泡颗粒细胞,分为模拟物转染组、抑制物转染组、模拟物阴性对照转染组、抑制物阴性对照转染组,分别转染miR-34a模拟物、miR-34a抑制物、模拟物阴性对照和抑制物阴性对照,另设空白对照组不做任何处理;实时定量荧光PCR(qRT-PCR)检测各组细胞miR-34a、Bcl-2、Bax、生存素、半胱氨酸天冬氨酸特异性蛋白酶-3(Caspase-3)的mRNA表达水平,蛋白质印迹法检测Bcl-2、Bax、生存素、Caspase-3、剪切体Caspase-3等凋亡相关蛋白的表达;流式细胞术检测细胞凋亡率。结果:miR-34a模拟物转染明显下调Bcl-2 mRNA表达水平,降低Bcl-2、生存素蛋白表达,并增加Bax、Caspase-3蛋白表达,促进人颗粒细胞凋亡;miR-34a抑制物转染作用则与之相反;然而,miR-34a模拟物和抑制物转染对Bax、生存素、Caspase-3的mRNA水平均无明显影响。结论:miR-34a可通过调控凋亡相关的蛋白与基因的表达,促进人颗粒细胞凋亡。
[Abstract]:Objective: to investigate the effect of miR-34a on human ovarian granulosa cells and its mechanism. Methods: human follicular granulosa cells were cultured in vitro and divided into three groups: mimic transfection group, inhibitor transfection group and mimic negative control group. In the negative control group, the miR-34a mimic was transfected with miR-34a inhibitor, the mimic negative control group and the inhibitor negative control group, and the blank control group was set up without any treatment. Real-time quantitative fluorescence PCR qRT-PCR was used to detect Bcl-2 Bax. survivin. The mRNA expression level of cysteine aspartate specific protease-3 (caspase-3) was detected by Western blot method. Bcl-2 Baxand survivin Caspase-3 were detected by Western blotting. The expression of apoptosis-related proteins such as Caspase-3; Results the expression of Bcl-2 mRNA, Bcl-2, survivin protein and Bax were significantly down-regulated by the transfection of the mimic of 1: miR-34a by flow cytometry. The expression of Caspase-3 protein promotes the apoptosis of human granulosa cells. The transfection effect of miR-34a inhibitor was opposite. However, miR-34a mimics and inhibitors were transfected into Bax-survivin. Conclusion Caspase-3 can promote apoptosis of human granulosa cells by regulating the expression of apoptosis-related proteins and genes.
【作者单位】：镇江市妇幼保健院生殖医学中心;
【基金】：国家自然科学基金资助项目(81402165) 江苏省卫生和计划生育委员会项目(H201451) 镇江市社会发展项目(SH2015057)
【分类号】：R714.8
【正文快照】： 颗粒细胞是支持卵细胞发育的重要细胞类型,影响卵泡启动、发育、成熟及闭锁的全过程,其过度凋亡阻止雄激素向雌激素转化,引起卵泡内局部激素环境变化,干扰正常卵泡发育,阻碍卵母细胞成熟,诱导卵泡闭锁,降低卵母细胞的质量及胚胎发育潜能,直接影响辅助生殖技术助孕的人群体外授

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