| 朱继荣,杨波,张博媛.miR-451a对慢性心衰细胞模型心肌细胞H9c2凋亡与自噬的影响.[J].中南医学科学杂志.,2023,(5):650-654. |
| miR-451a对慢性心衰细胞模型心肌细胞H9c2凋亡与自噬的影响 |
| Effects of miR-451a on apoptosis and autophagy of cardiomyocytes H9c2 in chronic heart failure |
| 投稿时间：2022-11-23 修订日期：2023-07-09 |
| DOI：10.15972/j.cnki.43-1509/r.2023.05.006 |
| 中文关键词: miR-451a mTOR信号通路 心肌细胞 凋亡 自噬 心衰 [ |
| 英文关键词:miR-451a mTOR signaling pathway cardiomyocyte apoptosis autophagy heart failure |
| 基金项目:广东省基础与应用基础研究基金项目(2021A1515010696) |
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| 中文摘要: |
| 目的探究miR-451a对慢性心衰细胞模型心肌细胞H9c2凋亡与自噬的作用机制。 方法采用1 μmol/L阿霉素(DOX)建立慢性心衰细胞模型。将细胞实验分为Control组(正常心肌细胞H9c2)、DOX组(DOX培养)、DOX+miR-NC组(DOX培养转染对照miR-NC)、DOX+miR-451a组(DOX培养转染miR-451a模拟物miR-451a)、DOX+miR-451a+CCI-779组(DOX和CCI-779培养转染miR-451a)。qRT-PCR检测各组细胞miR-451a相对表达水平；CCK-8和流式细胞术分别检测细胞增殖活性与凋亡；Western blotting检测剪切的Caspase-3(cleaved Caspase-3)、Bcl-2相关X蛋白(Bax)、Beclin-1、微管相关蛋白1轻链3I(LC3I)、LC3II、哺乳动物雷帕霉素靶蛋白(mTOR)、p70s6k、磷酸化mTOR(p-mTOR)、p-p70s6k蛋白表达。 结果成功建立慢性心衰细胞模型。与Control组比较,DOX组miR-451a相对表达水平、细胞活性、p-mTOR、p-p70s6k蛋白表达降低,而凋亡率、cleaved-Caspase-3、Bax、Beclin-1、LC3II/I蛋白表达升高。与DOX+miR-NC组比较,DOX+miR-451a组miR-451a相对表达水平、细胞活性、p-mTOR、p-p70s6k蛋白表达升高,而凋亡率、cleaved-Caspase-3、Bax、Beclin-1、LC3II/I蛋白表达降低。与DOX+miR-451a组比较,DOX+miR-451a+CCI-779组细胞活性降低,凋亡率、cleaved-Caspase-3、Bax、Beclin-1、LC3II/I蛋白表达升高。 结论miR-451a通过抑制mTOR信号通路促进慢性心衰细胞模型心肌细胞H9c2凋亡和自噬。 |
| 英文摘要: |
| AimTo explore the effects of miR-451a on apoptosis and autophagy of cardiomyocytes H9c2 in chronic heart failure. MethodsChronic heart failure cell model was constructed with 1 μmol/L doxorubicin (DOX). The cell experiments were divided into control group (normal myocardial cells H9c2), DOX group (DOX culture), DOX+miR-NC group (DOX culture transfected with miR-NC), DOX+ miR-451a group (DOX culture transfected with miR-451a mimic miR-451a) and DOX+miR-451a+CCI-779 group (DOX and CCI-779 culture transfected with miR-451a). The relative expression level of miR-451a was detected by qRT-PCR. The activity of cell proliferation and apoptosis were detected by CCK-8 and flow cytometry, respectively. The expressions of cleaved Caspase-3 (cleaved-Caspase-3), Bcl-2-associated X protein (Bax), Beclin-1, microtubule-associated protein 1 light chain 3I (LC3I), LC3II, phosphorylated mammalian target of rapamycin (p-mTOR), mammalian target of rapamycin (mTOR), p-p70s6k and p70s6k proteins were detected by Western blotting. ResultsThe model of chronic heart failure was successfully constructed. Compared with Control group, relative expression level of miR-451a, cell activity, expressions of p-mTOR and p-p70s6k proteins were significantly decreased, while apoptosis rate, expressions of cleaved-Caspase-3, Bax, Beclin-1 and LC3II/I proteins were significantly increased in DOX group. Compared with DOX+miR-NC group, relative expression level of miR-451a, cell activity, expressions of p-mTOR and p-p70s6k proteins were significantly increased, while apoptosis rate, expressions of cleaved-Caspase-3, Bax, Beclin-1 and LC3II/I proteins were significantly decreased in DOX+miR-451a group. Compared with DOX+miR-451a group, cell activity was significantly decreased, while apoptosis rate, expressions of cleaved-Caspase-3, Bax, Beclin-1 and LC3II/I proteins were significantly increased in DOX+miR-451a+CCI-779 group. ConclusionmiR-451a can promote apoptosis and autophagy of cardiomyocytes in chronic heart failure by inhibiting mTOR signaling pathway. |
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