Department of Three Cardiovascular Ward, The Second Affiliated Hospital of Shaanxi University of Traditional Chinese Medicine, Xianyang, Shaanxi, China.
Eur Rev Med Pharmacol Sci. 2018 Sep;22(18):6109-6118. doi: 10.26355/eurrev_201809_15950.
DJ-1-phosphate and tension homology deleted on chromosome ten/phosphatidylinositol-3-kinase/protein kinase B (PTEN/PI3K/AKT) signaling pathway plays a role in the regulation of ischemic reperfusion (I-R) injury. Bioinformatics analysis demonstrated that there is a complementary binding site between microRNA-192 (miR-192) and the 3'-UTR of DJ-1 mRNA. This study investigated the role of miR-192 in regulating DJ-1-PTEN/PI3K/AKT signaling pathway and myocardial I-R injury.
miR-122 and DJ-1 mRNA expressions in myocardial tissue were detected by Real-time PCR (RT-PCR). DJ-1, PTEN, and phosphorylated AKT (p-AKT) protein expressions were tested by Western blot. Reactive oxygen species (ROS) content was measured by flow cytometry. Malondialdehyde (MDA) content and superoxide dismutase (SOD) enzyme activity were detected by the kits. I-R treatment was performed at 72 h after transfection. Cell apoptosis was evaluated with flow cytometry.
Compared with sham group, miR-192, PTEN expressions and MDA content were significantly increased (p<0.05), while DJ-1, p-AKT levels and SOD activities were significantly reduced (p<0.05) in myocardial tissue of I-R group. Compared with control, I-R treatment significantly up-regulated miR-192 level, significantly decreased DJ-1 and p-AKT proteins, significantly elevated PTEN expression, and significantly induced apoptosis and ROS production in H9C2 cells (p<0.05). Transfection of miR-192 inhibitor significantly enhanced DJ-1 level, declined PTEN expression, elevated p-AKT level, and restrained apoptosis, ROS production and MDA content, and promoted SOD activity in H9C2 cells under I-R condition.
The expression of miR-192 increased significantly, while the expression of DJ-1 reduced obviously during I-R injury after myocardial infarction. Down-regulation of miR-192 markedly enhanced DJ-1 expression and PTEN/PI3K/AKT pathway activity, inhibited cell apoptosis and ROS generation, and reduced I-R injury in cardiomyocytes.
DJ-1 磷酸化和张力同源缺失于第十号染色体/磷酸肌醇-3-激酶/蛋白激酶 B（PTEN/PI3K/AKT）信号通路在调节缺血再灌注（I-R）损伤中发挥作用。生物信息学分析表明，微小 RNA-192（miR-192）和 DJ-1 mRNA 的 3'-UTR 之间存在互补结合位点。本研究探讨了 miR-192 在调节 DJ-1-PTEN/PI3K/AKT 信号通路和心肌 I-R 损伤中的作用。
通过实时 PCR（RT-PCR）检测心肌组织中 miR-122 和 DJ-1 mRNA 的表达。通过 Western blot 检测 DJ-1、PTEN 和磷酸化 AKT（p-AKT）蛋白的表达。通过流式细胞术测定活性氧（ROS）含量。通过试剂盒检测丙二醛（MDA）含量和超氧化物歧化酶（SOD）酶活性。转染 72 小时后进行 I-R 处理。通过流式细胞术评估细胞凋亡。
与假手术组相比，I-R 组心肌组织中 miR-192、PTEN 表达和 MDA 含量显著升高（p<0.05），而 DJ-1、p-AKT 水平和 SOD 活性显著降低（p<0.05）。与对照组相比，I-R 处理显著上调 miR-192 水平，显著下调 DJ-1 和 p-AKT 蛋白，显著上调 PTEN 表达，并显著诱导 H9C2 细胞凋亡和 ROS 产生（p<0.05）。转染 miR-192 抑制剂后，I-R 条件下 H9C2 细胞中的 DJ-1 水平显著升高，PTEN 表达下调，p-AKT 水平升高，凋亡、ROS 产生和 MDA 含量抑制，SOD 活性增强。
心肌梗死后 I-R 损伤时，miR-192 表达明显增加，DJ-1 表达明显降低。下调 miR-192 可显著增强 DJ-1 表达和 PTEN/PI3K/AKT 通路活性，抑制细胞凋亡和 ROS 生成，减轻心肌细胞 I-R 损伤。
