Downregulation of microRNA-126 in endothelial progenitor cells from diabetes patients, impairs their functional properties, via target gene Spred-1
Downregulation of microRNA-126 in endothelial progenitor cells from diabetes patients, impairs their functional properties, via target gene Spred-1
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糖尿病患者内皮祖细胞中 microRNA-126 的下调，通过靶基因 Spred-1 损害其功能特性
DOI：
10.1016/j.yjmcc.2012.04.003
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发表时间：
2012-07-01
影响因子：
5
通讯作者：
Wang, C. -Q.
中科院分区：
文献类型：
--
作者：
Meng, S.;Cao, J. -T.;Wang, C. -Q.
糖尿病(DM)对循环内皮祖细胞的数量和功能有不利影响。因此，这些细胞的修复机制也会减少，这是糖尿病血管疾病发展的关键和启动因素。本研究的目的是分析糖尿病患者内皮祖细胞miR的表达谱，选择最受调控的miR，以研究其在EPC功能障碍中的可能作用，并阐明其作用机制。内皮祖细胞来自II型糖尿病患者和非糖尿病对照组。从内皮祖细胞中提取总RNA，通过基因芯片筛选获得5个候选miRNAs，并用TaqMan实时定量聚合酶链式反应进行定量。将表达miR-126和miR-126抑制剂的慢病毒载体(抗miR-126)导入内皮祖细胞，检测内皮祖细胞的集落形成能力、增殖活性、迁移活性、分化能力和凋亡敏感性，并进行Western blotting和mRNA实时定量聚合酶链式反应分析。为了研究其作用机制，制备了表达Spred-1的慢病毒载体和针对Spred-1的siRNA。糖尿病患者内皮祖细胞中有5个MIR异常下调。这些miR包括miR-126、miR-21、miR-27a、miR-27b和miR-130a。抗miR-126能抑制内皮祖细胞的增殖、迁移和促进细胞凋亡。糖尿病大鼠内皮祖细胞miR-126表达恢复，促进内皮祖细胞增殖、迁移，抑制内皮祖细胞凋亡能力。尽管如此，miR-126对EPC的分化没有影响。MIR-126过表达显著下调内皮祖细胞中Spred-1的表达。下调糖尿病内皮祖细胞Spred-1的表达，可促进细胞增殖、迁移，抑制细胞凋亡。MiR-126影响内皮祖细胞的信号通路部分是通过调节Ras/ERK/VEGF和PI3K/Akt/eNOS来实现的。本研究首次提出糖尿病患者内皮祖细胞miR-126表达下调的证据，并通过其靶点Spred-1、Ras/ERK/VEGF和PI3K/Akt/eNOS信号通路损害内皮祖细胞的功能。(C)爱思唯尔有限公司出版的2012年。
Diabetes mellitus (DM) adversely affects the number and function of circulating endothelial progenitor cells (EPCs). Consequently, there is also a reduction in the repair mechanism of these cells, which is a critical and initiating factor in the development of diabetic vascular disease. The aim of the present study was to analyze miR expression profiles in EPCs from patients with DM and choose the most significantly regulated miR to study its possible role on EPC dysfunction and elucidate its mechanism of action. EPCs were collected from subjects with Type II DM and non-diabetic control subjects. Total RNA was harvested from EPCs, and a total of 5 candidate miRNAs were identified by microarray screening and were quantified by TaqMan real-time PCR. Lentiviral vectors expressing miR-126 and miR-126 inhibitor (anti-miR-126) were transfected into EPCs, and the EPC colony-forming capacity, proliferation activity, migratory activity, differentiation capacity, and apoptotic susceptibility were determined and Western Blotting and mRNA real-time PCR analyses were performed. To study the mechanisms, lentiviral vectors expressing Spred-1 and a short interfering RNA (siRNA) targeting Spred-1 were prepared. Five miRs were aberrantly downregulated in EPCs from DM patients. These miRs included miR-126, miR-21, miR-27a, miR-27b and miR-130a. Anti-miR-126 inhibited EPC proliferation, migration, and enhanced apoptosis. Restored miR-126 expression in EPCs from DM promoted EPC proliferation, migration, and inhibited EPC apoptosis ability. Despite this, miR-126 had no effect on EPC differentiation. miR-126 overexpression significantly downregulated Spred-1 in EPCs. The knockdown of Spred-1 expression in EPCs from DM promoted proliferation, migration, and inhibited apoptosis of the cells. The signal pathway of miR-126 effecting on EPCs is partially mediated through Ras/ERK/VEGF and PI3K/Akt/eNOS regulation. This study provides the first evidence that miR-126 is downregulated in EPCs from diabetic patients, and impairs EPCs-mediated function via its target, Spred-1, and through Ras/ERK/VEGF and PI3K/Akt/eNOS signal pathway. (c) 2012 Published by Elsevier Ltd.