Downregulation of miR-200a protects cardiomyocyte against apoptosis
Downregulation of miR-200a protects cardiomyocyte against apoptosis
复制标题
miR-200a 下调可保护心肌细胞免于凋亡
DOI：
10.1016/j.biopha.2019.109303
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发表时间：
2020-03-01
影响因子：
7.5
通讯作者：
Bai, Yunlong
中科院分区：
文献类型：
--
作者：
Wang, Yansong;Jiang, Yanan;Bai, Yunlong
背景/目标：急性心肌梗死（AMI）是缺血性心脏病的主要临床表现，是发病率和死亡率的重要原因。然而，缺血性心脏病发病机制中的关键调节因子仍存在争议。本研究旨在探讨miR-200 a在急性心肌梗死（AMI）中的作用及其机制。TTC染色测定梗死面积。采用H2 O2体外诱导AMI模型。将miR-200 a模拟物、抗miR-200 a反义寡脱氧核糖核苷酸（AMO-200 a）以及相应的阴性对照转染到心肌细胞中以观察miR-200 a的作用。流式细胞术检测细胞凋亡。实时荧光定量PCR、免疫荧光和蛋白质印迹法分别在RNA和蛋白质水平检测心肌细胞凋亡基因的表达。miR-200 a在小鼠心肌梗死周围区和H2 O2处理的心肌细胞中的表达均上调。AMO-200 a联合应用可减少凋亡细胞的数量，并改变凋亡相关蛋白的表达。有趣的是，生物信息学分析结果显示，miR-200 a可以与Fus mRNA的3 '-非翻译区（3'-UTR）结合。此外，Fus的表达在AMI小鼠模型和H2 O2处理的心肌细胞中下调。miR-200 a的改变负调控心肌细胞中Fus的表达。结论：MiR-200 a依赖的细胞凋亡信号通路在AMI损伤的发病机制中起重要作用，可能是一个激动人心的潜在治疗靶点。
Background/aims: Acute myocardial infarction (AMI) is a major clinical manifestation of ischemic heart disease and represents a significant cause of morbidity and mortality. However, key regulators in the pathogenesis of ischemic heart disease remain controversial. The present study was designed to investigate the involvement of miR-200a and its related mechanism in AMI.Methods: Left coronary artery (LCA) ligation was conducted to induce an AMI mouse model. The infarct size was measured by TTC staining. H2O2 was used to induce an AMI model in vitro. miR-200a mimics, anti-miR-200a antisense oligodeoxyribonucleotides (AMO-200a), as well as corresponding negative controls were transfected into cardiomyocytes to observe the effect of miR-200a. Flow cytometry was used to detect cell apoptosis. Real-time PCR, immunofluorescence and western blot assays were used to evaluate gene expression at RNA or protein levels, respectively.Results: Apoptosis was activated in AMI models. The expression of miR-200a was upregulated both in the peri-infarcted region of mice myocardium and H2O2-treated cardiomyocytes. The co-administration of AMO-200a decreased the number of apoptosis cells and altered the expression of apoptosis related proteins. Interestingly, bioinformatics analysis results revealed that miR-200a could bind to the 3'-untranslated regions (3'-UTR) of Fus mRNA. In addition, the expression of Fus was downregulated in the AMI mouse models and in H2O2-treated cardiomyocytes. The alteration of miR-200a negatively regulated Fus expression in cardiomyocytes. Also, the protective effect of AMO-200a was observed through its regulation of Fus.Conclusion: MiR-200a-dependent apoptosis signaling pathway plays an important role in the pathogenesis of AMI injury and could be an exciting potential therapeutic target.