含Ⅰ型血小板结合蛋白基序的解聚蛋白样金属蛋白酶10在压力超负荷诱导的心肌肥厚中的表达及其与心肌纤维化的关系  

Expression of a Disintegrin and Metalloproteinase With Thrombospondin-like Motifs 10 in Pressure Overload Induced Cardiac Hypertrophy and Its Relationship With Myocardial Fibrosis

作　　者：王震[1] 王梦龙[1] 刘剑芳[1] 叶晶[1] 徐瑶[1] 姜慧敏[1] 叶迪 张记收 万军[1] WANG Zhen;WANG Menglong;LIU Jianfang;YE Jing;XU Yao;JIANG Huimin;YE Di;ZHANG Jishou;WAN Jun(Department of Cardiology, Reiunin Hospital of Wuhan University, Cardiovascular Research Institute of Wuhan University, Hubei Key Laboratory of Cardiology, Wuhan ( 430060 ), Hubei, China)

机构地区：[1]武汉大学人民医院心血管内科

出　　处：《中国循环杂志》2019年第8期810-816,共7页Chinese Circulation Journal

基　　金：国家自然科学基金(81170208)

摘　　要：目的:探讨含Ⅰ型血小板结合蛋白基序的解聚蛋白样金属蛋白酶10(ADAMTS10)在压力超负荷诱导的心肌肥厚中的表达及其与心肌纤维化的关系。方法:将60只8周雄性C57BL/6J小鼠随机分成假手术组(n=20)和实验组(n=40)。应用主动脉弓缩窄术(TAC)建立压力负荷诱导的心肌肥厚模型。两组小鼠分别于第4周和第8周超声检测小鼠心功能,之后处死测量体质量和心脏质量,同时行心肌组织苏木素-伊红、天狼猩红、麦胚凝集素(WGA)和免疫荧光染色。应用免疫印迹(Westernblotting)法检测小鼠心肌组织ADAMTS10蛋白质的表达,应用实时聚合酶联反应(RT-PCR)法检测小鼠心肌组织ADAMTS10、心房钠尿肽(ANP)、β-肌球蛋白重链(β-MHC)、基质金属蛋白酶(MMP)2和MMP9mRNA的表达。结果:实验组与假手术组比较,小鼠第4周心脏质量显著升高(P<0.05),且在实验组第8周进一步升高(P<0.05);小鼠第4周左心室射血分数以及短轴缩短率均明显降低(P<0.05,且在第8周进一步降低,P<0.05);ADAMTS10mRNA和蛋白表达升高(P均<0.05)。免疫荧光结果提示ADAMTS10定位于成纤维细胞;RT-PCR结果提示ADAMTS10mRNA水平与心肌组织ANP水平显著正相关[r=0.6017,P<0.05(实验组4周)和r=0.6607,P<0.05(实验组8周)],与β-MHC水平显著正相关[r=0.6249,P<0.05(实验组4周)和r=0.7194,P<0.05(实验组8周)],与MMP2显著正相关[r=0.6971,P<0.05(实验组4周)和r=0.8689,P<0.05(实验组8周)],与MMP9水平显著正相关[r=0.6931,P<0.05(实验组4周)和r=0.8096,P<0.05(实验组8周)]。结论:在压力超负荷诱导的心肌肥厚模型,ADAMTS10通过调控细胞外基质重构,参与心肌肥厚和纤维化的发生发展。Objectives:To investigate the expression of a disintegrin and metalloproteinase with thrombospondin-like motifs 10(ADAMTS10) in cardiac hypertrophy induced by pressure overload and its relationship with myocardial fibrosis in mice.Methods:Eight weeks old C57 BL/6 J mice were randomly divided into sham group(n=20) and transverse aortic constriction(TAC) group(n=40).Echocardiography examination was performed at 4 or 8 weeks post-surgery,mice were sacrificed after echocardiographic examination respectively.The body weight and heart weight were measured.Meanwhile,heart sections were stained with hematoxylin eosin and Sirius red,wheat germ agglutinin(WGA) and immunofluorescence staining.The protein expression of ADAMTS10 protein in myocardium was detected by Western blotting.The mRNA expression of ADAMTS10,atrial natriuretic peptide(ANP),beta-myosin heavy chain(β-MHC),matrix metalloproteinases(MMP)2 and MMP9 in myocardium was detected by Real time-PCR.Results:Compared with the sham group,the heart weight was significantly increased in TAC 4-week group(P<0.05)and increased further in the TAC 8-week group(P<0.05).Compared with the sham group,left ventricular ejection fraction and fractional shortening were significantly decreased in TAC 4-week group(P<0.05),and further decreased in the TAC8-week group(all P<0.05).The myocardial mRNA and protein expression of ADAMTS10 in TAC groups was significantly upregulated as compared to sham group(P<0.05).Immunofluorescence results showed that ADAMTS10 was localized in cardiac fibroblasts.RT-PCR results indicated the there was a significant positive correlation between myocardial ADAMTS10 and ANP [r=0.6017,P<0.05(TAC 4 W) and r=0.6607,P<0.05(TAC 8 W)],between myocardial ADAMTS10 and β-MHC[r=0.6249,P<0.05(TAC 4 W) and r=0.7194,P<0.05(TAC 8 W)],between myocardial ADAMTS10 and MMP2[r=0.6971,P<0.05(TAC 4 W) and r=0.8689,P<0.05(TAC 8 W)],between myocardial ADAMTS10 and MMP9 [r=0.6931,P<0.05(TAC4 W) and r=0.8096,P<0.05(TAC 8 W)].Conclusions:The expression of ADAMTS10 is increased in mi

关 键 词：心肌肥厚 心肌纤维化 含Ⅰ型血小板结合蛋白基序的解聚蛋白样金属蛋白酶10 细胞外基质 

分 类 号：R541[医药卫生—心血管疾病]