MicroRNA-125b Affects Vascular Smooth Muscle Cell Function by Targeting Serum Response Factor
MicroRNA-125b Affects Vascular Smooth Muscle Cell Function by Targeting Serum Response Factor
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MicroRNA-125b 通过靶向血清反应因子影响血管平滑肌细胞功能
DOI：
10.1159/000489203
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发表时间：
2018-01-01
影响因子：
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通讯作者：
Chang, Guangqi
中科院分区：
文献类型：
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作者：
Chen, Zhibo;Wang, Mian;Chang, Guangqi
背景/目的：越来越多的证据表明microRNA与外周血管疾病的发病机制有关。我们最近发现microRNA-125 b（miR-125 b）是下肢动脉硬化闭塞症（阿索）患者动脉中最显著下调的microRNA之一。然而，其在阿索过程中的作用尚不清楚。本研究旨在探讨miR-125 b在ASO发生发展过程中的表达、调控机制及功能。方法：采用组织块贴壁法培养血管平滑肌细胞。建立大鼠颈动脉球囊损伤模型以模拟血管新生内膜的发展，并使用慢病毒转导系统过表达血清反应因子（SRF）或miR-125 b。采用实时荧光定量PCR（qRT-PCR）检测miR-125 b和SRF mRNA的表达水平。Western blotting检测SRF和Ki 67的表达水平。采用原位杂交技术分析miR-125 b的定位和表达水平。CCK-8和EdU测定用于评估细胞增殖，并且进行transwell和伤口闭合测定以测量细胞迁移。流式细胞术用于评估细胞凋亡，并进行双荧光素酶报告基因测定以检查miR-125 b对SRF的影响。采用免疫组化和免疫荧光法分析SRF和Ki 67的定位和表达水平。结果：在阿索动脉和血小板衍生生长因子（PDGF）BB刺激的VS MCs中，miR-125 b表达降低。miR-125 b抑制VSMC增殖和迁移，但促进VSMC凋亡。SRF被确定为是miR-125 b的直接靶标。外源性miR-125 b表达可调节球囊损伤大鼠颈动脉SRF表达并抑制血管新生内膜形成。结论：这些发现证明了miR-125 b/SRF通路在调节VSMC功能中的特定作用，并表明调节miR-125 b水平可能是治疗阿索的新方法。(C)2018作者由S Karger AG，巴塞尔出版
Background/Aims: Increasing evidence links microRNAs to the pathogenesis of peripheral vascular disease. We recently found microRNA-125b (miR-125b) to be one of the most significantly down-regulated microRNAs in human arteries with arteriosclerosis obliterans (ASO) of the lower extremities. However, its function in the process of ASO remains unclear. This study aimed to investigate the expression, regulatory mechanisms, and functions of miR-125b in the process of ASO. Methods: Using the tissue explants adherent method, vascular smooth muscle cells (VSMCs) were prepared for this study. A rat carotid artery balloon injury model was constructed to simulate the development of vascular neointima, and a lentiviral transduction system was used to overexpress serum response factor (SRF) or miR-125b. Quantitative real-time PCR (qRT-PCR) was used to detect the expression levels of miR-125b and SRF mRNA. Western blotting was performed to determine the expression levels of SRF and Ki67. In situ hybridization analysis was used to analyze the location and expression levels of miR-125b. CCK-8 and EdU assays were used to assess cell proliferation, and transwell and wound closure assays were performed to measure cell migration. Flow cytometry was used to evaluate cell apoptosis, and a dual-luciferase reporter assay was conducted to examine the effects of miR-125b on SRF. Immunohistochemistry and immunofluorescence analyses were performed to analyze the location and expression levels of SRF and Ki67. Results: miR-125b expression was decreased in ASO arteries and platelet-derived growth factor (PDGF) BB-stimulated VS MCs. miR-125b suppressed VSMC proliferation and migration but promoted VSMC apoptosis. SRF was determined to be a direct target of miR-125b. Exogenous miR-125b expression modulated SRF expression and inhibited vascular neointimal formation in balloon injured rat carotid arteries. Conclusions: These findings demonstrate a specific role of the miR-125b/SRF pathway in regulating VSMC function and suggest that modulating miR-125b levels might be a novel approach for treating ASO. (C) 2018 The Author(s) Published by S Karger AG, Basel