Chen Bairong, Luo Liyun, Zhu Weiping, Wei Xiaoliang, Li Songbiao, Huang Yin, Liu Mao, Lin Xiufang
Department of Cardiology, The Fifth Affiliated Hospital of Sun Yat-sen University, Zhuhai, Guangdong, China.
Medicine (Baltimore). 2016 Aug;95(33):e4418. doi: 10.1097/MD.0000000000004418.
The aim of this study is to determine miR-22 expression levels in peripheral blood mononuclear cells (PBMCs) of patients with coronary artery disease (CAD) and to investigate whether MCP-1 expression is regulated by miR-22. miR-22 expression in PBMCs from 60 CAD patients including stable angina pectoris (SAP) (n = 29), unstable angina pectoris (UAP) or non-ST elevation myocardial infarction (NSTEMI) (n = 17), or ST-elevation MI (STEMI) (n = 14) and 20 non-CAD subjects by real-time polymerase chain reaction (qRT-PCR). The luciferase activity assays were employed to determine whether miR-22 binds to 3'UTR of MCP-1. miR-22 mimics and inhibitors were transfected into healthy PBMCs. MCP-1 mRNA and protein levels were determined by qRT-PCR and enzyme-linked immuno sorbent assay, respectively. The qRT-PCR results showed that miR-22 levels in PBMCs were decreased in CAD patients, and MCP-1 was augmented in CAD patients and was inversely correlated with miR-22 levels. The luciferase activity assays indicated that MCP-1 was a target of miR-22. Overexpression of miR-22 could significantly repress MCP-1 expression at both mRNA and protein levels in PBMCs, whereas inhibition of miR-22 showed the opposite effects. This study revealed that miR-22 is downregulated in PBMCs from patients with CAD and that miR-22 may participate in inflammatory response by targeting MCP-1, therefore contributing CAD.
本研究旨在测定冠心病(CAD)患者外周血单个核细胞(PBMCs)中miR-22的表达水平,并研究MCP-1表达是否受miR-22调控。采用实时聚合酶链反应(qRT-PCR)检测60例CAD患者(包括稳定型心绞痛(SAP)患者29例、不稳定型心绞痛(UAP)或非ST段抬高型心肌梗死(NSTEMI)患者17例、ST段抬高型心肌梗死(STEMI)患者14例)及20例非CAD受试者PBMCs中miR-22的表达。采用荧光素酶活性测定法确定miR-22是否与MCP-1的3'UTR结合。将miR-22模拟物和抑制剂转染至健康PBMCs中。分别通过qRT-PCR和酶联免疫吸附测定法测定MCP-1 mRNA和蛋白水平。qRT-PCR结果显示,CAD患者PBMCs中miR-22水平降低,CAD患者中MCP-1升高,且与miR-22水平呈负相关。荧光素酶活性测定表明MCP-1是miR-22的靶标。miR-22过表达可显著抑制PBMCs中MCP-1在mRNA和蛋白水平的表达,而抑制miR-22则产生相反的效果。本研究表明,CAD患者PBMCs中miR-22表达下调,miR-22可能通过靶向MCP-1参与炎症反应,从而促进CAD的发生。