miR-181a-5p在ACS患者血清中表达及对人HCASMC细胞增殖迁移的调控作用、与ADAMTS1靶向关系  

Expression of miR-181a-5p in serum of ACS patients and its regulatory effects on proliferation and migration of human HCASMC cells,and its targeting relationship with ADAMTS1

作　　者：颚璐莎 易华 张艳芳 E Lusha;YI Hua;ZHANG Yanfang(不详;Department of Cardiology,Inner Mongolia Autonomous Region People's Hospital,Hohhot 010017,China)

机构地区：[1]内蒙古自治区人民医院心内科,呼和浩特010017 [2]内蒙古自治区人民医院病理科

出　　处：《山东医药》2024年第6期19-23,共5页Shandong Medical Journal

基　　金：内蒙古自治区卫生健康科技计划项目(202201014)。

摘　　要：目的观察微小RNA-181a-5p(miR-181a-5p)在急性冠脉综合征(acute coronary syndrome,ACS)患者血清中的表达及对人冠状动脉平滑肌细胞(human coronary artery smooth muscle cell,HCASMC)增殖和迁移的调控作用,探讨miR-181a-5p与血小板反应蛋白解整合素金属肽酶1(ADAMTS1)的靶向关系。方法采集100例ACS患者[ACS组,其中急性心肌梗死(AMI)患者55例、不稳定心绞痛(unstable angina pectoris,UAP)患者44例]、40例冠脉造影结果阴性者(对照组)的外周静脉血,采用实时荧光定量PCR法检测两组血清miR-181a-5p。取对数生长期HCASMC细胞,分为一、二、三、四组,采用脂质体转染法分别转染miR-181a-5p模拟物(miR-181a-5p mimics)、模拟物阴性对照(mimics-NC)、miR-181a-5p抑制物(miR-181a-5p inhibitor)及抑制物阴性对照(inhibitor-NC),培养48 h时采用CCK-8法测算细胞增殖能力、采用Transwell迁移实验测算细胞迁移能力,培养24 h时采用采用Western Blotting法检测各组细胞ADAMTS1蛋白。取对数生长期HCASMC细胞分为A、B、C、D四组,A组先后转染突变型ADAMTS1荧光素酶报告基因质粒(ADAMTS1-MUT)、miR-181a-5p mimics;B组先后转染ADAMTS1-MUT、mimics-NC;C组先后转染野生型ADAMTS1荧光素酶报告基因质粒(ADAMTS1-WT)、miR-181a-5p mimics;D组先后转染ADAMTS1-WT、mimics-NC,培养24 h时取各组细胞采用双荧光素酶报告基因实验检测各组细胞相对荧光素酶活性。结果与对照组相比,ACS组患者血清miR-181a-5p相对表达量低(P<0.01);与UAP患者相比,AMI患者血清miR-181a-5p相对表达量低(P<0.01)。与二组相比,培养48 h时一组细胞增殖能力和迁移能力降低,培养24 h时一组细胞ADAMTS1蛋白相对表达量低(P均<0.01);与四组相比,培养48 h时三组细胞增殖能力和迁移能力高,培养24 h时三组细胞ADAMTS1蛋白相对表达量高(P均<0.01)。A、B、C、D组细胞荧光素酶活性分别为1.03±0.03、1.01±0.04、0.45±0.06、1.02±0.05;与B组相比,A组细胞荧光素�Objective To investigate the expression of microRNA-181a-5p(miR-181a-5p)in the serum of patients with acute coronary syndrome(ACS)and its regulatory effects on the proliferation and migration of human coronary artery smooth muscle cells(HCASMC),and to explore the targeting relationship between miR-181a-5p and a disintegrin and me‐talloproteinasewith thrombospondin motifs 1(ADAMTS1).Methods Peripheral venous blood was collected from 100 patients with ACS[ACS group,including 55 cases of acute myocardial infarction(AMI)and 44 cases of unstable angina pectoris(UAP)]and 40 cases of negative coronary angiography(control group).Real-time quantitative PCR was used to detect the serum miR-181a-5p in the two groups.HCASMC in the logarithmic growth phase were divided into groups I,II,III and IV,which were transfected with miR-181a-5p mimics,mimics-NC,miR-181a-5p inhibitor,and inhibitor-NC by Liposome transfection method.The cell proliferation ability was measured by CCK-8 at 48 h of culture,and the cell migra‐tion ability was measured by Transwell migration experiment.At 24 h of culture,the ADAMTS1 protein was detected by Western blotting.HCASMC in the logarithmic growth phase were divided into groups A,B,C and D.Cells in the group A were transfected with mutant ADAMTS1 luciferase reporter gene plasmid(ADAMTS1-MUT)and miR-181a-5p mimics;cells in the group B were transfected with ADAMTS1-MUT and mimics-NC;cells in the group C were transfected with wild-type ADAMTS1 luciferase reporter gene plasmid(ADAMTS1-WT)and miR-181a-5p mimics;cells in the group D were transfected with ADAMTS1-WT and mimics-NC,respectively.The relative luciferase activity of cells in each group was de‐tected by dual luciferase reporter gene assay at 24 h after culture.Results Compared with the control group,the relative expression of serum miR-181a-5p was lower in the ACS group(P<0.01);compared with UAP patients,the relative ex‐pression of serum miR-181a-5p in AMI patients was lower(P<0.01).Compared with group II,the proliferation and migra‐tio

关 键 词：微小RNA-181a-5p 急性冠脉综合征 细胞增殖 细胞迁移 血小板反应蛋白解整合素金属肽酶1 

分 类 号：R34[医药卫生—基础医学]