半胱氨酸丰富跨膜成骨蛋白调控因子1-骨形态发生蛋白4信号通路对血管紧张素Ⅱ诱导的心室肌细胞肥大的调控作用研究  

Study on the regulation effect of cysteine-rich transmembrane bone morphogenetic protein regulator 1-bone morphogenetic protein 4 on ventricular myocyte hypertrophy induced by angiotensinⅡ

作　　者：李语 杨龙[2] 何炯红[2] 欧巧巧 杨英[2] LI Yu;YANG Long;HE Jiong-hong;OU Qiao-qiao;YANG Ying(Graduate School of Guizhou Medical University,Guiyang 550002,China;Department of Cardiology,Guizhou Provincial People’s Hospital,Guiyang 550002,China)

机构地区：[1]贵州医科大学研究生院,贵州省贵阳市550025 [2]贵州省人民医院心内科

出　　处：《中国心血管病研究》2024年第12期1125-1130,共6页Chinese Journal of Cardiovascular Research

基　　金：贵州省科学技术基金(黔科合基础-ZK[2021]一般360);贵州省卫生健康委科学技术基金(jzwkj2022-043)。

摘　　要：目的探讨半胱氨酸丰富跨膜成骨蛋白调控因子1(Crim1)是否通过抑制骨形态发生蛋白4(BMP4)而调节血管紧张素Ⅱ(AngⅡ)诱导的心室肌细胞肥大。方法分离1 d龄SD大鼠乳鼠心室肌获心室肌细胞,按干预方式不同分九组:Ad-Null(腺病毒空载体)组、Ad-Null+AngⅡ、Ad-Null+AngⅡ+Nog(重组人头蛋白)组、Ad-Crim1(Crim1基因重组腺病毒载体)组、Ad-Crim1+AngⅡ组、Ad-Crim1+AngⅡ+Nog组、Ad-shCrim1(shRNA干扰介导沉默Crim1基因重组腺病毒载体)组、Ad-shCrim1+AngⅡ组和Ad-shCrim1+AngⅡ+Nog组。实时荧光定量逆转录PCR检测β-肌球蛋白重链(β-MHC)和BMP4的mRNA表达。Western免疫印迹法检测全细胞提取蛋白BMP4的表达。细胞经结晶紫染色后通过Image J软件测量细胞表面积。结果AngⅡ干预明显增加心室肌细胞β-MHC mRNA表达以及细胞面积,上调BMP4 mRNA/蛋白表达;Ad-Crim1和Nog干预明显抑制AngⅡ的上述效应。Ad-shCrim1干预沉默Crim1基因对心室肌细胞β-MHC mRNA表达,细胞面积及BMP4 mRNA/蛋白表达与AngⅡ干预效应相似,该效应为Nog干预明显抑制。结论Crim1通过下调BMP4的表达而抑制AngⅡ诱导的乳鼠心室肌细胞肥大。Objective To investigate whether cysteine rich transmembrane osteogenic protein regulatory factor 1(Crim1)regulates angiotensinⅡ(AngⅡ)-induced ventricular myocyte hypertrophy by inhibiting bone morphogenetic protein 4(BMP4).Methods The ventricular myocytes from 1-day-old SD rats were isolated and divided into nine groups according to different intervention methods:Ad-Null(adenovirus empty vector)group,Ad-Null+AngⅡgroup,Ad-Null+AngⅡ+Nog(recombinant human noggin)group,Ad-Crim1(recombinant adenovirus vector of Crim1 gene)group,Ad-Crim1+AngⅡgroup,Ad-Crim1+AngⅡ+Nog group,Ad-shCrim1(shRNA interference mediated silencing of Crim1 gene recombination adenovirus vector)group,Ad-shCrim1+AngⅡgroup and Ad-shCrim1+AngⅡ+Nog group.The mRNA expression of myosin heavy chain beta(β-MHC)and BMP4 was detected by RT-qPCR.The protein level of BMP4 in the whole-cell extracts was determined by Western blot analysis.The cells stained with crystal violet were measured for cell surface area using Image J software.Results AngII intervention significantly increasedβ-MHC mRNA expression,BMP4 mRNA/protein expression and cell area of ventricular myocytes;the above effects of AngII intervention were significantly inhibited by pre-using of Ad-Crim1 and Nog.Crim1 gene of ventricular myocytes silenced by Ad-shCrim1 intervention leading to the similar effects of AngⅡintervention onβ-MHC mRNA expression,BMP4 mRNA/protein expression and cell area,those were significantly inhibited by Nog intervention.Conclusions Crim1 inhibits AngⅡ-induced hypertrophy of neonatal rat ventricular myocytes by decreasing BMP4 expression.

关 键 词：心肌肥大 半胱氨酸丰富跨膜成骨蛋白调控因子1 骨形态发生蛋白4 血管紧张素Ⅱ 

分 类 号：Q95-33[生物学—动物学] R541.4[医药卫生—心血管疾病]