MicroRNA-143 promotes cardiac ischemia-mediated mitochondrial impairment by the inhibition of protein kinase Cepsilon
MicroRNA-143 promotes cardiac ischemia-mediated mitochondrial impairment by the inhibition of protein kinase Cepsilon
复制标题
MicroRNA-143 通过抑制蛋白激酶 Cepsilon 促进心肌缺血介导的线粒体损伤
DOI：
10.1007/s00395-017-0649-7
复制
发表时间：
2017-11-01
影响因子：
9.5
通讯作者：
Zhang, Rong
中科院分区：
文献类型：
--
作者：
Hong, Hong;Tao, Ting;Zhang, Rong
蛋白激酶Cepsilon(PKC Epsilon)对心肌梗死(MI)的保护作用是通过其抗细胞凋亡的特性和microRNA(MiRNA)靶向调控的潜在机制来实现的。利用动物心肌梗死和细胞缺氧模型，结合一系列先进的分子技术，对MI诱导的损伤、PKCE的表达以及miRNA-143(miR-143)对PKCE的靶向调节进行了评估。结果表明，随着心肌梗死面积的增加以及细胞的凋亡和坏死损伤，PKCE的表达显著下调，并建立了miR-143与PKCE之间的靶向关系和潜在的结合图谱。体内和体外缺血实验均显示miR-143可诱导细胞凋亡和坏死，而AnagomiR-143或AMO-143可逆转这一作用。在体外，miR-143基因的转染上调miR-143细胞的表达也会导致细胞损失，而miR-143与AMO-143共转染后这种作用被完全逆转。在动物心肌梗死和细胞缺氧模型中，也观察到miR-143对线粒体膜电位和ATP合成的同样的有害作用，以及miR-143过表达并被antagomiR或AMO转换的模型。重要的是，在所有测试的模型中，miR-143的过表达下调了PKCE，并且这种下调在antagomiR或AMO的存在下被逆转。荧光素酶报告基因和miRNA掩蔽实验证实miR-143对PKCE具有直接靶向调控作用。综上所述，MI介导的miR-143上调抑制了PKCE的表达，从而干扰了PKCE对线粒体的心肌保护作用，最终导致线粒体膜电位耗散和心肌死亡。
The cardioprotection of protein kinase Cepsilon (PKC epsilon) against myocardial infarction (MI) mediated by its anti-apoptotic property and underlying mechanism of targeted regulation by microRNA (miRNA) are not established. MI-induced injury, PKCe expression, and targeted regulation of miRNA-143 (miR-143) to PKCe have been evaluated using animal MI and cellular hypoxic models conjugated with series of state-of-art molecular techniques. The results demonstrated that PKCe significantly downregulated along with increased infarcted area and apoptotic and necrotic damage in MI model, and the targeted relationship and potential binding profile were established between miR-143 and PKCe. Both in vivo and in vitro ischemic tests showed that miR-143 induced apoptosis and necrosis, which was reversed by antagomiR-143 or AMO-143. The upregulation of miR-143 by transfection of miR-143 in vitro also induced cell loss, and this effect of miR-143 was completely reversed by co-transfection of miR-143 with AMO-143. The identically deleterious action of miR-143 on mitochondrial membrane potential and ATP synthesis was also observed in both animal MI and cellular hypoxic models, as well as miR-143 overexpressed models and converted by either antagomiR or AMO. Importantly, overexpression of miR-143 downregulated PKCe in all tested models and this downregulation was reversed in the presence of antagomiR or AMO. The direct targeted regulation of miR-143 on PKCe was confirmed by luciferase reporter and miRNA-masking tests. In conclusion, MI-mediated upregulation of miR-143 inhibits PKCe expression and consequently interference with the cardioprotection of PKCe to mitochondrial, and leads to mitochondrial membrane potential dissipation and myocardial death eventually.