miR-182-5p促进心肌梗死大鼠心肌细胞凋亡的机制研究  

Mechanism of miR-182-5p Promoting Myocardial Cell Apoptosis in Rats with Myocardial Infarction

作　　者：裴亚军 王艳红[1] PEI Yajun;WANG Yanhong(Shanxi Medical University,Taiyuan 030001,Shanxi,China;Third Hospital of Shanxi Medical University(Shanxi Bethune Hospital,Shanxi Academy of Medical Sciences,Tongji Shanxi Hospital),Taiyuan 030032,Shanxi,China)

机构地区：[1]山西医科大学,太原030001 [2]山西医科大学第三医院(山西白求恩医院山西医学科学院同济山西医院),太原030032

出　　处：《中西医结合心脑血管病杂志》2024年第17期3143-3150,3157,共9页Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease

摘　　要：目的:探究miR-182-5p对心肌细胞凋亡的影响,进而以更好的阐明miR-182-5p在心肌梗死中的作用和意义。方法:构建大鼠心肌梗死模型,末端脱氧核苷酸转移酶介导的dUTP缺口末端标记测定法(TUNEL)法检测心肌凋亡、实时荧光定量逆转录聚合酶链式反应(qRT-PCR)及蛋白免疫印迹(Western Blot)法检测miR-182-5p及凋亡相关分子的表达。使用miR-182-5p mimics及inhibitor转染大鼠心肌细胞H9c2,构建缺氧模型,细胞计数试剂盒-8(CCK-8)法检测心肌细胞活力,Western Blot法检测凋亡相关指标表达水平,半胱氨酰蛋白酶3/7(Caspase-3/7)活性检测试剂盒检测Caspase-3/7的活性。使用pAAV9-CMV腺相关病毒构建miR-182-5p的过表达病毒,通过尾静脉注射至心肌梗死模型小鼠体内,检测AAV9-miR-182-5p对心肌梗死诱导的心肌凋亡水平的影响。结果:大鼠心肌梗死模型中miR-182-5p表达水平减少,且细胞凋亡水平增加。在H9c2心肌缺氧模型中,miR-182-5p可以加重缺氧对心肌细胞活力的抑制,并减少抗凋亡分子B细胞淋巴瘤2(Bcl-2)、增加促凋亡分子Bcl-2相关X蛋白(Bax)的表达,同时对Caspase-3/7的活性有明显的促进作用。而AAV9-miR-182-5p的注射可以显著加重小鼠心肌梗死模型中心肌细胞的凋亡水平。结论:miR-182-5p可以加重心肌梗死下的心肌损伤,其途径是通过促进凋亡来实现的。Objective:To investigate the mechanism of miR-182-5p promoting myocardial cell apoptosis in rats with myocardial infarction.Methods:Myocardial infarction model of rats was constructed.Myocardial apoptosis was detected by terminal deoxyribonucleotidyl transferase(TdT)-mediated dUTP nick end labeling(TUNEL).The expression of miR-182-5p and apoptosis-related molecules were detected by real-time quantitative fluorescent reverse transcription polymerase chain reaction(qRT-PCR)and Western Blot.miR-182-5p mimics and inhibitor transfected rat cardiomyocytes H9c2 to construct hypoxia model.Cell count Kit 8(CCK-8)was used to detect cardiomyocyte cell viability.The expression levels of apoptosis-related factor were detected by Western Blot.Caspase-3/7 activity detection kit was used to detect the activity of Caspase-3/7.The overexpression virus of miR-182-5p was constructed using pAAV9-CMV adeno-associated virus and injected into the mouse model of myocardial infarction through the tail vein to detect the effect of AAV9-miR-182-5p on the level of myocardial apoptosis induced by myocardial infarction.Results:The expression level of miR-182-5p decreased and the apoptosis level increased in the rat model of myocardial infarction.In H9c2 myocardial hypoxia model,miR-182-5p could aggravate the inhibition of hypoxia on cardiomyocyte viability,reduce the anti-apoptotic molecule B-cell lymphoma 2(Bcl-2),and increase the expression of pro-apoptotic molecule Bcl-2 associated X protein(Bax).At the same time,the activity of Caspase-3/7 was significantly promoted.The injection of AAV9-miR-182-5p significantly increased the level of apoptosis of cardiomyocytes in mouse myocardial infarction models.Conclusion:miR-182-5p can aggravate myocardial damage after myocardial infarction by promoting apoptosis.

关 键 词：心肌梗死 miR-182-5p 心肌缺氧 细胞凋亡 实验研究 

分 类 号：R542.22[医药卫生—心血管疾病]