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高血压诱导血管损伤的分子机制:微小RNA-34a的表达及作用

Molecular mechanisms in vascular injury induced by hypertension: Expression and role of microRNA-34a.

作者信息

Liu Siguan, Yi Fanfan, Cheng Wenwei, Qu Xin, Wang Chunting

机构信息

Department of Critical Care Medicine, Shandong Provincial Hospital Affiliated to Shandong University, Jinan, Shandong 250101, P.R. China.

Emergency Department, Zaozhuang Municipal Hospital, Zaozhuang, Shandong 277101, P.R. China.

出版信息

Exp Ther Med. 2017 Dec;14(6):5497-5502. doi: 10.3892/etm.2017.5216. Epub 2017 Sep 27.

Abstract

The aim of the present study was to investigate the expression and function of microRNA (miR)-34a in patients with primary hypertension. The expression of miR-34a was measured in the peripheral blood of 50 patients with primary hypertension and 28 normal controls by reverse transcription quantitative polymerase chain reaction. In addition, human umbilical vein endothelial cells (HUVECs) were transfected with an miR-34a inhibitor to suppress the expression of miR-34a, and the proliferation, migration and cell cycle distribution of HUVECs were measured by Cell Counting Kit-8, Transwell and flow cytometry assays. The target of miR-34a was also predicted by bioinformatics analysis and verified by a dual-luciferase reporter gene assay and western blot analysis. miR-34a was significantly upregulated in the peripheral blood of patients with hypertension when compared with controls (P<0.05), and upregulation of miR-34a was associated with a higher clinical stage of hypertension (phase III; P<0.05). experiments demonstrated that inhibition of miR-34a promoted the proliferation, migration and G1/S transition of HUVECs, relative to scramble-miR controls (P<0.05). Furthermore, transforming growth factor β-induced factor homeobox 2 (TIGF2) was predicted and verified to be a direct target of miR-34a. Collectively, these data suggested that miR-34a was upregulated in the peripheral blood of patients with hypertension, and that upregulated miR-34a may promote vascular endothelial injury by targeting TIGF2.

摘要

本研究的目的是调查微小RNA(miR)-34a在原发性高血压患者中的表达及功能。采用逆转录定量聚合酶链反应检测50例原发性高血压患者和28例正常对照者外周血中miR-34a的表达。此外,用miR-34a抑制剂转染人脐静脉内皮细胞(HUVECs)以抑制miR-34a的表达,通过细胞计数试剂盒-8、Transwell和流式细胞术检测HUVECs的增殖、迁移及细胞周期分布。通过生物信息学分析预测miR-34a的靶标,并通过双荧光素酶报告基因检测和蛋白质免疫印迹分析进行验证。与对照组相比,高血压患者外周血中miR-34a显著上调(P<0.05),且miR-34a的上调与高血压的更高临床分期(Ⅲ期;P<0.05)相关。实验表明,相对于乱序miR对照组,抑制miR-34a可促进HUVECs的增殖、迁移及G1/S期转变(P<0.05)。此外,预测并验证转化生长因子β诱导因子同源盒2(TIGF2)是miR-34a的直接靶标。总的来说,这些数据表明高血压患者外周血中miR-34a上调,且上调的miR-34a可能通过靶向TIGF2促进血管内皮损伤。

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