miR-143在人颈动脉粥样硬化患者斑块组织中表达变化及作用机制  

Expression of miR-143 in plaque tissue of patients with carotid atherosclerosis and its mechanism

作　　者：宋坚 王先法[1] 朱越锋[1] Song Jian;Wang Xianfa;Zhu Yuefeng(Department of General Surgery,Sir Run Run Shaw Hospital,School of Medicine,Zhejiang University,Hangzhou 310016,Zhejiang;Department of General Surgery,Ningbo fourth hospital,Ningbo 315700,Zhejiang)

机构地区：[1]浙江大学医学院附属邵逸夫医院普外科,杭州310016 [2]宁波市第四医院普外科,宁波315700

出　　处：《中国组织化学与细胞化学杂志》2019年第6期508-513,共6页Chinese Journal of Histochemistry and Cytochemistry

基　　金：浙江省医药卫生科技项目(2018ZH016)。

摘　　要：目的探讨miR-143在人颈动脉粥样硬化患者斑块组织中的表达变化及可能的作用机制。方法收集我院血管外科因颈动脉狭窄行颈动脉内膜剥脱术(CEA)切除的颈动脉斑块组织及斑块旁内膜组织56例,根据术前颈动脉超声检查及术后斑块病理学检查将所取得的斑块组织又分为稳定斑块组26例和不稳定斑块组30例。另外选择同期年龄、性别相匹配的在我院因外伤、门脉高压或原发性血小板增多症而行脾脏切除术患者的正常的脾动脉内膜组织20例作为正常对照组,通过实时荧光定量PCR(qRT-PCR)检测组织中miR-143的表达水平。体外培养人主动脉血管平滑肌细胞(HAVSMCs),分别转染mi R-143模拟物、模拟物阴性对照、miR-143抑制物、抑制物阴性对照,采用qRT-PCR检测转染后HAVSMC细胞miR-143表达水平的变化,CCK-8法和Transwell迁移实验分别检测转染后细胞增殖活性和迁移能力的变化。通过生物信息学预测miR-143的可能靶基因,双荧光素酶报告基因实验观察miR-143与ADAMTS4的靶向调控关系,Western blot观察miR-143对ADAMTS4表达水平的影响。结果颈动脉粥样硬化斑块组织中miR-143表达水平明显低于斑块旁内膜组织和正常动脉血管内膜组织,并且不稳定斑块组患者斑块组织中miR-143的表达水平明显低于稳定斑块组斑块组织。转染miR-143模拟物后,HAVSMCs中miR-143表达水平明显升高,增殖活性和迁移能力明显降低;转染miR-143抑制物后,HAVSMCs中miR-143表达水平明显降低,增殖活性和迁移能力明显升高。双荧光素酶报告基因显示ADAMTS4是miR-143的直接靶基因,Western blot显示miR-143能够在转录后水平负向调控ADAMTS4表达。结论miR-143在颈动脉粥样硬化斑块组织中表达降低,其可能通过靶向调控ADAMTS4影响血管平滑肌细胞增殖和迁移能力,与颈动脉斑块的形成和不稳定性有关。Objective To investigate the expression of miR-143 in human carotid atherosclerosis plaque and its possible mechanism.Methods 56 cases of carotid plaque tissues and adjacent intima tissues resected by carotid endarterectomy(CEA)for carotid stenosis in our hospital were collected.According to preoperative carotid ultrasonography and postoperative plaque pathology examination,the obtained plaque tissues were divided into stable plaque group(26 cases)and unstable plaque group(30 cases).In addition,20 cases of normal splenic artery intima tissue of patients undergoing splenectomy due to trauma,portal hypertension or idiopathic thrombocytosis in our hospital were selected as normal control group.Quantitative Real-time fluorescence PCR(qRT-PCR)was used to detect the expression of miR-143 in these tissues.Human aortic vascular smooth muscle cells(HAVSMCs)were cultured in vitro and transfected with miR-143 mimics,mimics negative control,miR-143 inhibitor and inhibitor negative control,respectively.The expression of miR-143 in HAVSMC cells after transfection was detected by qRT-PCR.The proliferation activity and migration ability of HAVSMC cells after transfection were detected by CCK-8 method and transwell migration test,respectively.Bioinformatics was used to predict the possible target genes of miR-143,double luciferase reporter gene experiment was used to observe the targeting regulation relationship between miR-143 and ADAMTS4,and Western blot was used to observe the effect of miR-143 on the expression level of ADAMTS4 protein.Results The expression level of miR-143 in carotid atherosclerotic plaque tissue was significantly lower than that in para-plaque intima tissue and normal arterial intima tissue.The expression level of miR-143 was significantly lower in unstable plaque group than that in stable plaque group.After transfection of miR-143 mimics,the expression level of miR-143 in HAVSMC cells increased significantly,proliferation activity and migration ability decreased significantly;after transfection of miR-143

关 键 词：MIR-143 颈动脉粥样硬化斑块 增殖 迁移 ADAMTS4 

分 类 号：R543.4[医药卫生—心血管疾病]