心肌重构分子机制解析  

Analysis of Molecular Mechanism of Myocardial Remodeling

作　　者：王立英[1] 张馨予 刘家铭 吴昕嵘 张广超 杨明硕 周阳 张艳琦[2] WANG Li-Ying;ZHANG Xin-Yu;LIU Jia-Ming;WU Xin-Rong;ZHANG Guang-Chao;YANG Ming-Shuo;ZHOU Yang;ZHANG Yan-Qi(Basic Medicine College of Beihua University,Jilin 132013,China;Chinese Institute of Standardization,Beijing 100191,China;College of Chemistry&Chemical Engineering,Lanzhou University,Lanzhou 730000,China)

机构地区：[1]北华大学基础医学院,吉林132013 [2]中国标准化研究院,北京100191 [3]兰州大学化学化工学院,兰州730000

出　　处：《分析化学》2022年第5期711-719,共9页Chinese Journal of Analytical Chemistry

基　　金：吉林省教育厅“十三五”科学技术研究项目(No.JJKH20220067KJ);吉林省科技发展计划项目(No.20190303038SF);吉林市科技发展计划项目(No.20190302018);吉林省卫生健康委员会项目(No.2019J051)资助。

摘　　要：考察了在体及离体情况下多种信号分子在心肌重构(Myocardial remodeling,MR)中的作用,并基于多种检测技术探讨了其作用机制。首先应用异丙肾上腺素(Isoprenaline,Iso)建立了Wistar大鼠MR的在体模型,并通过血管紧张素Ⅱ(AngiotensinⅡ,AngⅡ)诱导新生乳鼠心肌成纤维细胞(Myocardial fibroblasts,myoFbs)增殖,导致心肌纤维化(Myofibrosis,MF),从而建立了MR离体模型。进一步,基于Western blot技术进行相关蛋白表达检测以及甲基偶氮唑盐(Methyl thiazolyl tetrazolium,MTT)法进行的体外myoFbs增殖检测,结果表明,Iso可明显增加心肌组织中羟脯氨酸(Hydroxyproline,Hyp)含量和蛋白激酶Cα(Protein kinase Cα,PKCα)的表达。研究发现,10^(-7)mol/L AngⅡ即可显著促进myoFbs增殖;进一步应用免疫细胞化学染色及激光共聚焦显微镜方法进行p-PKCα、细胞外信号调节蛋白激酶(Extracellular signal regulated protein kinases,ERKl/2)及活性氧(Reactive oxygen species,ROS)在myoFbs中的表达及含量检测,结果表明,MR的形成与ROS、p-PKCα和p-ERKl/2表达量增多有关。The roles of various signaling molecules in myocardial remodeling(MR)in vivo and in vitro were investigated and the mechanisms based on the combination of various detection techniques was explored in this study.First,the in vivo MR model of Wistar rat was established using isoprenaline(Iso).AngiotensinⅡ(AngⅡ)induced proliferation of neonatal rat myofibroblasts(myoFbs)and thus set up the MR model in vitro.Further,Western blot was applied to detection of the expression of PKCα(protein kinase Ca,PKCα)in vivo,while methyl thiazolyl tetrazolium(MTT)assay was adopted for detection of myoFbs proliferation in vitro.Iso increased both the content of hydroxyproline(Hyp)and the expression of PKCαin myocardial tissue significantly.At the same time,it was found that AngⅡ(10^(-7)mol/L)promoted the proliferation of myoFbs significantly.Immunocytochemical staining and laser confocal microscopy were used to detect expression and content of p-PKCα,extracellular signal regulating protein kinase(ERKl/2)and reactive oxygen species(ROS)induced by AngⅡin myoFbs.The results showed that the formation of MR was related to the increase of expression of ROS,p-PKCαand p-ERKl/2.

关 键 词：心肌重构 心肌纤维化 心肌成纤维细胞 蛋白激酶CΑ 活性氧 

分 类 号：R542.2[医药卫生—心血管疾病]