Department of Pharmacology (State-Province Key Laboratories of Biomedicine-Pharmaceutics of China, Key Laboratory of Cardiovascular Medicine Research, Ministry of Education), College of Pharmacy, Harbin Medical University, Harbin, China.
Basic Res Cardiol. 2017 Sep 8;112(6):60. doi: 10.1007/s00395-017-0649-7.
The cardioprotection of protein kinase Cepsilon (PKCε) 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, PKCε expression, and targeted regulation of miRNA-143 (miR-143) to PKCε have been evaluated using animal MI and cellular hypoxic models conjugated with series of state-of-art molecular techniques. The results demonstrated that PKCε 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 PKCε. 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 PKCε in all tested models and this downregulation was reversed in the presence of antagomiR or AMO. The direct targeted regulation of miR-143 on PKCε was confirmed by luciferase reporter and miRNA-masking tests. In conclusion, MI-mediated upregulation of miR-143 inhibits PKCε expression and consequently interference with the cardioprotection of PKCε to mitochondrial, and leads to mitochondrial membrane potential dissipation and myocardial death eventually.
蛋白激酶 Cepsilon（PKCε）通过其抗凋亡特性对心肌梗死（MI）的心脏保护作用及其受 microRNA（miRNA）靶向调控的潜在机制尚未确定。使用动物 MI 和细胞缺氧模型，结合一系列最先进的分子技术，评估了 MI 诱导的损伤、PKCε 的表达以及 miRNA-143（miR-143）对 PKCε 的靶向调节。结果表明，PKCε 在 MI 模型中显著下调，同时梗死面积增加，凋亡和坏死损伤增加，并且建立了 miR-143 与 PKCε 之间的靶向关系和潜在结合模式。体内和体外缺血试验表明，miR-143 诱导细胞凋亡和坏死，而 antagomiR-143 或 AMO-143 逆转了这种作用。体外转染 miR-143 上调 miR-143 也诱导细胞丢失，而 miR-143 的这种作用被 miR-143 与 AMO-143 共转染完全逆转。在动物 MI 和细胞缺氧模型中，以及在 miR-143 过表达模型中和通过 antagomiR 或 AMO 转化的模型中，也观察到 miR-143 对线粒体膜电位和 ATP 合成的相同有害作用。重要的是，在所有测试模型中，miR-143 的过表达均下调 PKCε，并且在存在 antagomiR 或 AMO 的情况下，这种下调得到逆转。miR-143 对 PKCε 的直接靶向调节通过荧光素酶报告基因和 miRNA 掩蔽试验得到证实。总之，MI 介导的 miR-143 上调抑制 PKCε 的表达，从而干扰 PKCε 对线粒体的心脏保护作用，导致线粒体膜电位耗散和心肌死亡。
