心肌成纤维细胞DDR2对整合素β1调控的机制研究  

Regulation Mechanisms of DDR2 on the Expression of Integrinβ1 in the Cardiac Fibroblast

作　　者：郝春媛 李同华 李霞[2] 葛兴利 张誉洋 HAO Chunyuan;LI Tonghua;LI Xia(Department of Cardiovascular,The First Affiliated Hospital of Northwest University&No.1 Hospital of Xi'an City,Shaanxi 710002,China)

机构地区：[1]西北大学附属第一医院暨西安市第一医院心血管内科,710002 [2]西北大学附属第一医院暨西安市第一医院心血管研究室,710002 [3]西北大学附属第一医院暨西安市第一医院心脏重症监护室,710002

出　　处：《医学研究杂志》2023年第8期94-99,共6页Journal of Medical Research

基　　金：陕西省自然科学基础研究计划面上项目(2021-JM587)。

摘　　要：目的探究心肌成纤维细胞盘状结构域受体(discoid domain receptor 2,DDR2)对于整合素(integrin)β1表达的调控分子机制。方法体外培养成年SD大鼠心肌成纤维细胞,使用RNA干扰和高表达载体分别下调和上调DDR2的表达,Western blot法检测血管紧张素Ⅱ(angiogensinⅡ,AngⅡ)作用细胞后其整合素β1、细胞外信号调节蛋白激酶(extracellular signal-regulated kinase,Erk1/2)和转化生长因子-β1(transforming growth factor-β1,TGF-β1)的表达水平。其后使用Erk1/2的特异性抑制剂PD98059以及TGF-β1的RNA干扰阻断相应信号通路,Western blot法再次检测AngⅡ刺激细胞后整合素β1的表达变化。结果AngⅡ可以上调心肌成纤维细胞DDR2以及整合素β1的表达水平。DDR2的RNA干扰可以阻断AngⅡ引起的心肌成纤维细胞整合素β1的升高效应,同时降低了磷酸化Erk1/2(p-Erk1/2)和TGF-β1的表达水平。使用Erk1/2的特异性抑制剂PD98059可以削弱AngⅡ引起的心肌成纤维细胞TGF-β1和整合素β1的升高效应,而TGF-β1的RNA干扰可以削弱AngⅡ引起的心肌成纤维细胞整合素β1的升高效应。高表达DDR2可以显著性升高TGF-β1和整合素β1的表达水平,而TGF-β1的RNA干扰可以削弱DDR2上调引起的心肌成纤维细胞整合素β1的升高效应。结论AngⅡ通过DDR2-Erk1/2-TGF-β1信号途径调控心肌成纤维细胞整合素β1的表达水平,该信号途径可能在心肌损伤后结构改建中起着重要作用。Objective To investigate the regulatory molecular mechanism of discoid domain receptor(DDR2)on the expression of integrinβ1 in the cardiac fibroblasts.Methods Adult SD rat myocardial fibroblasts were cultured in vitro,and RNA interference and expression vector of DDR2 was transfected into the rat to downregulate or upregulate the expression of DDR2.Western blot was performed to detect the expression of integrinβ1,extracellular signal-regulated kinase(Erk1/2)and transforming growth factor-β1(TGF-β1)in the cardiac fibroblasts treated with angiogensinⅡ(AngⅡ).After that,the specific inhibitor of Erk1/2 PD98059 and RNA interference of TGF-β1 were used to block the corresponding signal pathway,Western blot was also performed to detect the expression changes of integrinβ1 in the cardiac fibroblasts treated with AngⅡ.Results AngⅡcould upregulate the expression levels of DDR2 and integrinβ1 in cardiac fibroblasts.RNA interference of DDR2 blocked the elevated effect of integrinβ1 in cardiac fibroblasts induced by AngⅡ,and decreased the expression levels of p-Erk1/2 and TGF-β1.It was shown that the specific inhibitor of Erk1/2 PD98059 attenuated the elevated effect of TGF-β1 and integrinβ1 in cardiac fibroblasts induced by AngⅡ,while RNA interference of TGF-β1 was able to attenuate the elevated effect of integrinβ1 in cardiac fibroblasts induced by AngⅡ.Upregulation of DDR2 expression was demonstrated to significantly increase the expression level of TGF-β1 and integrinβ1,while RNA interference of TGF-βI was able to attenuate the elevated effect of integrinβ1 in cardiac fibroblasts by DDR2 upregulation.Conclusion AngⅡis capable of regulating the expression level of integrinβ1 in cardiac fibroblasts through the DDR2-Erk1/2-TGF-βIsignaling pathway,which may play an important role in the ventricular remolding process after myocardial injury.

关 键 词：心肌成纤维细胞 盘状结构域受体2 整合素Β1 ERK1/2 TGF-Β1 

分 类 号：R541.6[医药卫生—心血管疾病]