Yes相关蛋白在血管紧张素Ⅱ诱导的心肌纤维化中的作用及机制研究  

Role of Yes-related protein in AngⅡ-induced myocardial fibrosis and its mechanism

作　　者：赵倩茹 曹梦菲 孙侠 钱晗 吕书梅 仲威[1] 邵晨[1] 王中群[1] 袁伟[1] Zhao Qianru;Cao Mengfei;Sun Xia;Qian Han;LüShumei;Zhong Wei;Shao Chen;Wang Zhongqun;Yuan Wei(Department of Cardiology,Affiliated Hospital of Jiangsu University,Zhenjiang 212001,Jiangsu Province,China)

机构地区：[1]江苏大学附属医院心血管内科,镇江212001

出　　处：《中华老年心脑血管病杂志》2022年第3期306-310,共5页Chinese Journal of Geriatric Heart,Brain and Vessel Diseases

基　　金：江苏省333工程科研项目(BRA2020395);江苏省重点研发计划项目(BE2021694)。

摘　　要：目的探讨Yes相关蛋白(YAP)在血管紧张素Ⅱ(AngⅡ)诱导的心肌纤维化中的作用及其机制。方法C57/B6小鼠30只,随机分为观察组、盐水组和模型组[腹腔注射AngⅡ1.5 mg/(kg·d)]4周,构建心肌纤维化模型,每组10只。将出生1~3 d SD乳鼠,差速贴壁法提取原代心肌成纤维细胞(CF),随机分为4组,对照组、AngⅡ组、抑制组(20μmol/L PD98059加1μmol/L AngⅡ)和DMSO组,均刺激细胞24 h。Western blot检测α平滑肌肌动蛋白(α-SMA)和磷酸化细胞外信号调节激酶(p-ERK)及YAP表达。免疫荧光观察α-SMA表达;CCK-8检测CF增殖活力;Transwell实验检测细胞迁移能力。结果与观察组相比,模型组小鼠p-ERK(1.02±0.09 vs 0.61±0.15)及YAP(1.00±0.10 vs 0.63±0.05)表达明显升高(P<0.01)。与对照组比较,AngⅡ组CF的α-SMA荧光强度、A值及迁移细胞明显升高(P<0.05);抑制组上述指标较AngⅡ组明显降低(P<0.05)。与对照组比较,AngⅡ组CF的p-ERK、总YAP、核YAP表达明显升高(P<0.05),抑制组CF的p-ERK(0.64±0.08 vs 1.12±0.09)、总YAP(0.67±0.06 vs 1.06±0.11)、核YAP(0.84±0.20 vs 1.32±0.12)表达较AngⅡ组明显降低(P<0.05,P<0.01)。结论AngⅡ可能通过激活ERK与YAP信号通路诱导心肌纤维化。Objective To study the role of Yes-related protein(YAP)in AngⅡ-induced myocardial fibrosis and its mechanism.Methods Thirty C57/B6 mice were randomly divided into observation group,normal saline group and model group(10 in each group).The myocardial fibrosis model was established by intraperitoneal injection with AngⅡ[1.5 mg/(kg·d)].Primary myocardial fibroblasts were divided into control group,AngⅡgroup,inhibition group and DMSO group,and simulated for 24 h after they were extracted from 1-3 days old SD mice and cultured with differential adherence method.The expressions ofα-SMA,p-ERK and YAP were detected by Western blot.The expression ofα-SMA was detected with immunoflrescence method.The proliferating activity of myocardial fibroblasts was assayed with CCK-8.The migration ability of myocardial fibroblasts was tested in Transwell experiment.Results The expression levels of p-ERK and YAP were significantly higher in model group than in observation group(1.02±0.09 vs 0.61±0.15,1.00±0.10 vs 0.63±0.05,P<0.01).The fluorescence intensity and A value were significantly higher and the number of migrated myocardial fibroblsts was significantly greater in AngⅡgroup than in control group(P<0.05).The fluorescence intensity and A value were significantly lower and the number of migrated myocardial fibroblsts was significantly smaller in inhibition group than in AngⅡgroup(P<0.05).The expression levels of p-ERK and YAP were sig-nificantly higher in AngⅡgroup than in control group(P<0.05)and significantly lower in inhi-bition group than in AngⅡgroup(0.64±0.08 vs 1.12±0.09,0.67±0.06 vs 1.06±0.11,0.84±0.20 vs 1.32±0.12,P<0.05,P<0.01).Conclusion AngⅡcan induce myocardial fibrosis by activating the ERK and YAP signaling pathways.

关 键 词：血管紧张素Ⅱ 心内膜心肌纤维化症 成纤维细胞 细胞运动 

分 类 号：R542.2[医药卫生—心血管疾病]