本文關(guān)鍵詞：miR-34a調(diào)節(jié)人卵巢顆粒細胞凋亡　出處：《江蘇大學學報(醫(yī)學版)》2016年06期 　論文類型：期刊論文

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【摘要】：目的:探討miR-34a對人卵巢顆粒細胞的作用及其機制。方法:體外培養(yǎng)人卵泡顆粒細胞,分為模擬物轉(zhuǎn)染組、抑制物轉(zhuǎn)染組、模擬物陰性對照轉(zhuǎn)染組、抑制物陰性對照轉(zhuǎn)染組,分別轉(zhuǎn)染miR-34a模擬物、miR-34a抑制物、模擬物陰性對照和抑制物陰性對照,另設(shè)空白對照組不做任何處理;實時定量熒光PCR(qRT-PCR)檢測各組細胞miR-34a、Bcl-2、Bax、生存素、半胱氨酸天冬氨酸特異性蛋白酶-3(Caspase-3)的mRNA表達水平,蛋白質(zhì)印跡法檢測Bcl-2、Bax、生存素、Caspase-3、剪切體Caspase-3等凋亡相關(guān)蛋白的表達;流式細胞術(shù)檢測細胞凋亡率。結(jié)果:miR-34a模擬物轉(zhuǎn)染明顯下調(diào)Bcl-2 mRNA表達水平,降低Bcl-2、生存素蛋白表達,并增加Bax、Caspase-3蛋白表達,促進人顆粒細胞凋亡;miR-34a抑制物轉(zhuǎn)染作用則與之相反;然而,miR-34a模擬物和抑制物轉(zhuǎn)染對Bax、生存素、Caspase-3的mRNA水平均無明顯影響。結(jié)論:miR-34a可通過調(diào)控凋亡相關(guān)的蛋白與基因的表達,促進人顆粒細胞凋亡。
[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.
【作者單位】： 鎮(zhèn)江市婦幼保健院生殖醫(yī)學中心;
【基金】：國家自然科學基金資助項目(81402165) 江蘇省衛(wèi)生和計劃生育委員會項目(H201451) 鎮(zhèn)江市社會發(fā)展項目(SH2015057)
【分類號】：R714.8
【正文快照】： 顆粒細胞是支持卵細胞發(fā)育的重要細胞類型,影響卵泡啟動、發(fā)育、成熟及閉鎖的全過程,其過度凋亡阻止雄激素向雌激素轉(zhuǎn)化,引起卵泡內(nèi)局部激素環(huán)境變化,干擾正常卵泡發(fā)育,阻礙卵母細胞成熟,誘導(dǎo)卵泡閉鎖,降低卵母細胞的質(zhì)量及胚胎發(fā)育潛能,直接影響輔助生殖技術(shù)助孕的人群體外授

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