Yin Jie, Liu Hongyan, Huan Lei, Song Suping, Han Liying, Ren Faxin, Zhang Zengtang, Zang Zhiqiang, Zhang Junye, Wang Shu
Department of Cardiology, Laiwu People's Hospital, Laiwu, Shangdong 271100, P.R. China.
Department of The Second Medicine, Laiwu People's Hospital, Laiwu, Shangdong 271100, P.R. China.
Exp Ther Med. 2017 Oct;14(4):2751-2756. doi: 10.3892/etm.2017.4886. Epub 2017 Aug 4.
The present study aimed to investigate the role and mechanism of micro RNA (miR)-128 in hypertension-induced myocardial injury. The peripheral blood of patients with hypertension was collected and the expression of miR-128 was detected using fluorescence reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Primary myocardial cells isolated from rat were cultured under conditions of hypoxia and glucose deprivation, and miR-128 expression was measured by RT-qPCR. The expression of c-Met protein was measured using western blot analysis and the apoptosis of transfected cells was measured by flow cytometry in rat myocardial cells following transfection with miR-128 mimics or c-Met siRNA. A luciferase assay was applied to assess the binding of miR-128 to c-Met mRNA. miR-128 expression was significantly higher in hypertension patients compared with controls (P<0.05). miR-128 expression was higher in patients with stage III/IV hypertension compared with patients with stage II hypertension. Similarly, miR-128 expression in primary cardiomyocytes cultured under deprivation of oxygen and glucose increased with the culture time and reached a peak at 12 h. c-Met expression decreased significantly (P<0.05) and the ratio of apoptotic cells increased significantly (P<0.05), following transfection of miR-128 mimics. The number of apoptotic cells also increased when c-Met expression was knocked down by siRNA. The dual luciferase assay indicated that fluorescence intensity decreased significantly in miR-128 mimics and wild type c-Met group (P<0.05), indicating that miR-128 can directly target c-Met. Therefore, the results of the current study suggest that miR-128 may promote myocardial cell injury by regulating c-Met expression.
本研究旨在探讨微小RNA（miR）-128在高血压所致心肌损伤中的作用及机制。收集高血压患者的外周血，采用荧光逆转录定量聚合酶链反应（RT-qPCR）检测miR-128的表达。分离大鼠原代心肌细胞，在缺氧和无糖条件下培养，用RT-qPCR检测miR-128的表达。用蛋白质免疫印迹法检测c-Met蛋白的表达，用流式细胞术检测转染miR-128模拟物或c-Met小干扰RNA（siRNA）后大鼠心肌细胞的凋亡情况。应用荧光素酶报告基因检测法评估miR-128与c-Met mRNA的结合情况。与对照组相比，高血压患者的miR-128表达显著升高（P<0.05）。与Ⅱ期高血压患者相比，Ⅲ/Ⅳ期高血压患者的miR-128表达更高。同样，在缺氧和无糖培养条件下的原代心肌细胞中，miR-128表达随培养时间增加而升高，并在12小时达到峰值。转染miR-128模拟物后，c-Met表达显著降低（P<0.05），凋亡细胞比例显著增加（P<0.05）。当用siRNA敲低c-Met表达时，凋亡细胞数量也增加。双荧光素酶报告基因检测表明，miR-128模拟物与野生型c-Met组的荧光强度显著降低（P<0.05），表明miR-128可直接靶向c-Met。因此，本研究结果提示，miR-128可能通过调节c-Met表达促进心肌细胞损伤。
