CTRP9对血管紧张素Ⅱ诱导的大鼠心肌成纤维细胞活化和增殖的影响  

Effects of CTRP9 on angiotensinⅡinduced rat cardiac fibroblasts activation and proliferation

作　　者：梁翠[1] 高路[1] 姚瑞[1] 李亚彭[1] 李鹏程 刘源[1] 张殿红[1] 王铮[1] 张彦周[1] LIANG Cui;GAO Lu;YAO Rui;LI Yapeng;LI Pengcheng;LIU Yuan;ZHANG Dianhong;WANG Zheng;ZHANG Yanzhou(Department of Cardiology,the First Affiliated Hospital,Zhengzhou University,Zhengzhou 450052)

机构地区：[1]郑州大学第一附属医院心血管内科,郑州450052

出　　处：《郑州大学学报（医学版）》2020年第2期228-232,共5页Journal of Zhengzhou University(Medical Sciences)

基　　金：国家自然科学基金项目(81600191)。

摘　　要：目的:探讨CTRP9对血管紧张素Ⅱ(AngⅡ)诱导的大鼠心肌成纤维细胞活化、增殖的影响。方法:分离培养原代大鼠心肌成纤维细胞,将其分为对照组、AngⅡ(1μmol/L)组和不同浓度(10、25、50、100、200 mg/L)CTRP9组,48 h后采用CCK-8法检测细胞增殖率。细胞分为对照组、AngⅡ(1μmol/L)组、50和200 mg/L CTRP9组,48 h后采用α-SMA染色观察成纤维细胞活化情况,qRT-PCR法检测Ⅰ型胶原、Ⅲ型胶原、纤维连接蛋白、结缔组织生长因子mRNA的表达,Western blot法检测AMPKa、P70和mTOR蛋白的磷酸化情况。采用AMPKa抑制剂复合物C处理细胞,将细胞分为对照组、AngⅡ(1μmol/L)组、CTRP9+复合物C(10μmol/L)组和CTRP9(200 mg/L)组,48 h后重复上述检测。结果:AngⅡ组心肌成纤维细胞增殖率高于对照组,各CTRP9组细胞增殖率低于AngⅡ组(P<0.05)。AngⅡ组细胞α-SMA表达高于对照组,50和200 mg/L CTRP9组α-SMA表达低于AngⅡ组。AngⅡ组细胞Ⅰ型胶原、Ⅲ型胶原、纤维连接蛋白和结缔组织生长因子mRNA的表达高于对照组,50和200 mg/L CTRP9组细胞Ⅰ型胶原、Ⅲ型胶原、纤维连接蛋白和结缔组织生长因子mRNA的表达低于AngⅡ组(P<0.05)。AngⅡ组细胞AMPKa蛋白磷酸化水平低于对照组,P70和mTOR蛋白磷酸化水平高于对照组;200 mg/L CTRP9组AMPKa蛋白磷酸化水平高于AngⅡ组,P70和mTOR蛋白磷酸化水平低于AngⅡ组(P<0.05)。结论:CTRP9通过激活AMPKa信号通路抑制AngⅡ诱导的大鼠心肌成纤维细胞活化、增殖功能。Aim:To investigate the effects of CTRP9 on angiotensinⅡ(AngⅡ)induced rat cardiac fibroblasts activation and proliferation.Methods:Primary cardiac fibroblasts were isolated and cultured.Cells were divided into control group,AngⅡ(1μmol/L)stimulation group and different concentrations of CTRP9(10,25,50,100,200 mg/L)treatment groups.After 48 hours,cell proliferation was detected by CCK-8 assay.Cells were divided into control group,AngⅡ(1μmol/L)group,50 and 200 mg/L CTRP9 groups.α-SMA expression was evaluated by immuno-staining to assess fibroblast activation.qRT-PCR was used to detect typeⅠcollagen,typeⅢcollagen,fibronectin and connective tissue growth factor mRNA.Western blot was used to detect AMPKa,P70,and mTOR protein.AMPKa inhibitor compound C,was used to reversed the phenotype.Cells were then divided into four groups:control group,AngⅡ(1μmol/L)group,CTRP9+compound C(10μmol/L)treatment group,and CTRP9(200 mg/L)treatment group.Then cell proliferation and activation were observed through the methods above.Results:The proliferation rate of cardiac fibroblasts in the AngⅡgroup was significantly higher than that in the control group(P<0.05).The proliferation rate of fibroblasts in 10,25,50,100,200 mg/L CTRP9 groups were lower than that in the AngⅡgroup(P<0.05).The expression ofα-SMA in AngⅡgroup was higher than that in control group,and the expression ofα-SMA in 50 and 200 mg/L CTRP9 groups was lower than that in AngⅡgroup.mRNA expressions of typeⅠcollagen,typeⅢcollagen,fibronectin and connective tissue growth factor were higher than those of the control group(P<0.05).mRNA expressions of typeⅠcollagen,typeⅢcollagen,fibronectin,and connective tissue growth factor in 50 and 200 mg/L CTRP9 groups were lower than those in AngⅡgroup(P<0.05).The expression of phosphorylated AMPKa protein in AngⅡgroup was lower than that in control group,and the expressions of phosphorylated P70 and mTOR protein were higher than those in control group(P<0.05).The expressions of phosphorylated AMPKa p

关 键 词：CTRP9 心肌成纤维细胞 心肌纤维化 血管紧张素Ⅱ AMPKa 大鼠 

分 类 号：R542.2[医药卫生—心血管疾病]