肝细胞生长因子和转化生长因子-β1对人心房成纤维细胞基质金属蛋白酶-2及组织抑制剂-2表达的影响  

Effects of hepatocyte growth factor and transforming growth factor-β1 on expression of matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-2 in human atrial fibroblasts

作　　者：陈建泉[1] 张建成[1] 陈林[1] 林亚洲[1] 

机构地区：[1]福建省立医院心血管内科 福建医科大学省立临床医学院,福州350001

出　　处：《中华心律失常学杂志》2017年第6期489-495,共7页Chinese Journal of Cardiac Arrhythmias

基　　金：福建省卫生厅重点创新项目（2007-CX-1）

摘　　要：目的 研究肝细胞生长因子（hepatocyte growth factor,HGF）和转化生长因子 β1（transforming growth factor-beta1,TGF-β1）对体外培养的人心房成纤维细胞基质金属蛋白酶-2（matrix metalloproteinase-2,MMP-2）及其组织抑制因子-2（tissue inhibitor of metalloproteinase-2,TIMP-2）表达的影响.方法 福建省立医院心外科住院行瓣膜置换术的风湿性心脏病（rheumatic heart disease,RHD）患者共20例,窦性心律（SR）和持续性心房颤动（CAF）患者各10例,记录患者临床资料.换瓣术中取适量右心耳,酶消化及差速贴壁法原代培养人心房成纤维细胞并免疫荧光鉴定,100 ng/ml HGF和10 ng/ml TGF-β1分别干预48 h并实验分组;用RT-PCR法检测人心房成纤维细胞MMP-2和TIMP-2 mRNA表达水平.结果 与SR组比较,CAF组患者左心房内径（left atrial diameter,LAD）扩大,SR组（4.43±0.77）cm,CAF组（5.76±1.37）cm,P〈0.05）;心房成纤维细胞MMP-2 mRNA表达增加,TIMP-2 mRNA减少（P〈0.01）;MMP-2 mRNA相对含量与LAD大小呈正相关（r=0.786）,与心房颤动（房颤）持续时间呈正相关（r=0.763）,与TIMP-2 mRNA相对含量呈负相关（r=-0.826）.与CAF组相比,TGF-β1组MMP-2 mRNA表达增加,TIMP-2 mRNA减少（P〈0.01）;TGF-β1＋HGF组MMP-2 mRNA表达降低,TIMP-2 mRNA升高（P〈0.05）;HGF组MMP-2 mRNA表达降低,TIMP-2 mRNA升高（P〈0.01）.与TGF-β1＋HGF组相比,HGF组MMP-2 mRNA相对含量减少,TIMP-2 mRNA增加（P〈0.05）;TGF-β1组MMP-2 mRNA相对含量增加,TIMP-2 mRNA减少（P〈0.01）.以上差异均有统计学意义.结论 RHD合并房颤患者LAD明显增大,MMP-2/TIMP-2表达比率增加;TGF-β1和HGF分别提高和减低人心房成纤维细胞MMP-2/TIMP-2表达率,参与促进或抑制房颤患者心房重构,HGF能够部分抑制TGF-β1的作用,有助于探索心房纤维化的分子机制.Objective To investigate the effects of hepatocyte growth factor（ HGF） and transforming growth factor beta1（ TGF-β1 ） on the expression of matrix metalloproteinase-2（ MMP-2） and tissue inhibitor of metalloproteinase-2（ TIMP-2） in cultured human atrial fibroblasts. Methods We enrolled 20 patients with rheumatic heart disease（ RHD） ,10 with sinus rhythm（ SR） and 10 with constant atrial fibrillation（ CAF） ,and clinical data of all patients were recorded.Right atrial appendage tissue was harvested from patients undergoing valve replacement.Primary human atrial fibroblasts were cultured with enzyme digestion and differential adherence method, and identified by immunofluorescence staining. 100 ng/ml HGF and 10 ng/ml TGF-β1 were respectively administered to cultured human atrial fibroblasts and incubated for 48 h,then divided into different experimental groups.The expressions of MMP-2 and TIMP-2 mRNA in human atrial fibroblasts were detected by RT-PCR. Results Compared with SR patients,CAF patients had enlarged left atrium diameter （LAD）[SR （4. 43±0. 77） cm vs.CAF （5. 76±1. 37） cm,P〈0. 05].The expression of MMP-2 mRNA increased and TIMP-2 mRNA decreased in atrial fibroblasts of CAF group（ P〈0. 01）;the relative content of MMP-2 mRNA was positively correlated with size of LAD（ r=0. 786） and duration of atrial fibrillation（ AF, r=0. 763）,and was negatively correlated with the relative content of TIMP-2 mRNA（r=-0. 826）.Compared with CAF group,the expression of MMP-2 mRNA increased and TIMP-2 mRNA decreased in TGF-β1 group （ P〈0. 01）; the expression of MMP-2 mRNA decreased and TIMP-2 mRNA increased in TGF-β1 with HGF intervention group （ P〈0. 05 ）; in HGF group, the expression of MMP-2 mRNA decreased and TIMP-2 mRNA increased（P〈0. 01）.Compared with TGF-β1 plus HGF group,the relative level of MMP-2 mRNA was lower and TIMP-2 mRNA was higher in HGF group（ P〈0. 05）;the content of MMP-2 mRNA was higher and TIMP-2 mRNA was lower in TGF-β1 g

关 键 词：肝细胞生长因子 转化生长因子-Β1 基质金属蛋白酶-2 金属蛋白酶组织抑制因子-2 

分 类 号：R541.2[医药卫生—心血管疾病] R541.75[医药卫生—内科学]