血管紧张素Ⅱ经AT_1受体和ERK1/2上调心肌细胞Cx43间隙连接  

AngiotensinⅡ up-regulates cardiomyocyte Cx43 gap expression via AT_1 receptor and ERK1/2

作　　者：王业松[1] 马虹[1] 胡苑[1] 曾武涛[1] 张惠忠[2] 陈健文[3] 

机构地区：[1]中山大学第一医院心内科 [2]中山大学肿瘤医院病理科,广东广州510080 [3]中山大学肿瘤医院生理实验科,广东广州510080

出　　处：《中国病理生理杂志》2006年第1期58-62,共5页Chinese Journal of Pathophysiology

基　　金：广州市科委基金资助项目(No.2002J1C0181)

摘　　要：目的:探讨血管紧张素Ⅱ对培养新生鼠心室肌细胞Cx43间隙连接的影响及其机制。方法:AngⅡ处理培养心肌细胞24h。缬沙坦、PD98059在AngⅡ刺激细胞前1h加到培养基中,对照组加等体积药物溶剂DMSO。用Westernblotting分析、代谢标记和免疫沉淀测定、电镜观察心肌细胞Cx43表达、合成和间隙连接。结果:Westernblotting分析显示用10-9-10-6mol/LAngⅡ刺激细胞24h,Cx43的表达与对照组相比呈浓度依赖性增加;用AngⅡ0.1μmol/L刺激心肌细胞24h,磷酸化ERK1/2(P-ERK1/2)活性高于对照组(P<0.01),AT1受体拮抗剂缬沙坦(1μmol/L)能完全阻断AngⅡ增加P-ERK1/2活性;用AngⅡ0.1μmol/L刺激心肌细胞24h,Cx43表达及P-ERK1/2活性均高于对照组(P<0.01),ERK1/2激酶特异性抑制剂PD98059(1μmol/L)能阻断AngⅡ上调Cx43表达和增加P-ERK1/2活性。代谢标记和免疫沉淀测定显示AngⅡ处理组放射渗入Cx43的量明显高于对照组(P<0.01),缬沙坦(1μmol/L)能完全阻断AngⅡ增加放射渗入Cx43的量。电镜观察表明用AngⅡ0.1μmol/L刺激心肌细胞24h,AngⅡ处理组细胞间隙连接数目和大小大于对照组(P<0.05)。结论:AngⅡ通过AT1受体和ERK1/2促进培养心肌细胞合成Cx43,上调Cx43表达和增加间隙连接数目及大小。AIM： To investigate the effect of angiotensin Ⅱ on Cx43 gap junction in cultured neonatal rat cardiac myocytes and its mechanism. METHODS： The cardiomyocytes were treated with AngⅡ for 24 h, which were pretreated with valsartan or PD98059 for 60 rain before Ang Ⅱ treatment. The controls were treated with equal amount of DMSO. The Cx43 expression, synthesis and gap junction in cardiomyocytes were characterized by Western blotting, metabolic labeling and immunoprecipitation assay, and electron microscope. RESULTS： Western blotting analysis revealed that Cx43 content concentration- dependently increased in cells treated with 10^-9 - 10^-6 mol/L AngⅡ for 24 h. Phosphorylated extracellular signal regulated kinase （P- ERK） 1/2 activity increased in cells treated with 0.1 μmol/L Ang Ⅱ for 24 h （ P 〈 0.01 vs control）, which was inhibited by the presence of 1 μmol/L valsartan, an AT1 - receptor blocker. Cx43 expression and P- ERK1/2 activity increased in cells treated with 0.1 μmol/L AngⅡ for 24 h （ P 〈 0.01 vs control）, which was inhibited by the presence of 1 μmol/L PD98059, a specific inhibitor of ERK1/2 kinase. Metabolic labeling and immunoprecipitation assay revealed that Ang Ⅱ increased the amount of radioactivity incorporated into Cx43, which was blocked by valsartan. Electron microscopy demonstrated increases in the number and size of gap junction profiles in cells exposed to 0.1 μmol/L AngⅡ for 24 h （ P 〈 0.05 vs control）. CONCLUSIONS： AngⅡ facilitates Cx43 synthesis , up - regulates Cx43 expression and increases the number and size of gap junction profiles in cultured neonatal rat cardiac myocytes via AT1 receptor and ERK1/2.

关 键 词：血管紧张素Ⅱ 心肌细胞 连接蛋白43 缝隙接合部 有丝分裂素激活蛋白激酶类 

分 类 号：R363[医药卫生—病理学]