AMPK激活在抑制大鼠心肌细胞肥大中的作用  

Effect of AMPK activation in the inhibition of hypertrophy of rat cardiomyocytes

作　　者：陈保林[1] 俞杉[1] 熊肇军[2] 张成喜[2] 马跃东[3] 陈广琴[3] 刘晨[3] 董吁钢[3] 

机构地区：[1]贵州省人民医院,贵阳550002 [2]中山大学附属第三医院 [3]中山大学附属第一医院

出　　处：《山东医药》2015年第10期17-20,共4页Shandong Medical Journal

基　　金：贵州省科技厅基金资助项目(黔科合J字[2011]2247号)

摘　　要：目的探讨单磷酸腺苷激活的蛋白激酶(AMPK)激活对抑制大鼠心肌细胞肥大的影响及其机制。方法分离培养新生大鼠心肌细胞,将培养的心肌细胞分为对照组、苯肾上腺素(PE)组、PA组、PAC组。PE组给予PE10μmol/L,PA组给予PE 10μmol/L及AMPK激活剂5-氨基-4-甲酰胺咪唑核糖核苷酸(AICAR)1 mmol/L,PAC组给予PE 10μmol/L、AICAR 1 mmol/L及AMPK抑制剂Compound C 1μmol/L。采用Western blot法检测各组AMPK表达,高效液相色谱仪检测各组心肌细胞三甲基组氨酸(3-MH)释放量,实时荧光定量PCR法检测各组ANP mRNA表达,采用免疫组化法测定各组心肌细胞表面积。结果 PE组3-MH释放量低于对照组,PA组3-MH释放量高于PE组,PAC组3-MH释放量低于PA组(P均<0.01)。PE组ANP mRNA表达高于对照组,PA组ANP mRNA表达低于PE组,PAC组ANP mRNA表达高于PA组(P均<0.01)。PE组心肌细胞表面积大于对照组,PA组心肌细胞表面积小于PE组,PAC组心肌细胞表面积大于PA组(P均<0.01)。结论 AMPK在心肌肥大发生过程中具有重要作用,激活AMPK能够促进心肌细胞蛋白质降解而逆转心肌肥大,抑制AMPK则可促进心肌肥大的发展。Objective To explore the effect of adenosine monophosphate-activated protein kinase （ AMPK） activation in the inhibition of hypertrophy of rat cardiomyocytes in vitro and to investigate the underlying mechanisms .Methods Neonatal SD rat cardiomyocytes were isolated and cultured .To observe the effect of 5-aminoimidazole-4-carboxamide-4-ri-bofuranoside （ AICAR） and Compound C on the activation of AMPK , the cultured cells were divided into three groups：the control group, AICAR group（AICAR 1 mmol/L） and AICAR ＋Compound C group （AICAR 1 mmol/L ＋Compound C 1μmol/L） .To examine the effect of AMPK activation on hypertrophy of rat cardiomyocytes , the cultured cardiomyocytes were divided into four groups： the control group, phenylephrine group （PE 10 μmol/L）, PA group （PE 10 μmol/L ＋AICAR 1 mmol/L） and PAC group （PE 10μmol/L ＋AICAR 1 mmol/L ＋Compound C 1 μmol/L）.AMPK and p-AMPK protein levels in cardiomyocytes were detected by using Western blotting .High performance liquid chromatographic detection was used to determine the 3-methylhistidine （3-MH） concentration in medium.The mRNA expression of fetal gene atrial natriuretic peptide （ ANP） was measured by real-time polymerase chain reaction （ real-time PCR） .and the sur-face areas of cardiomyocytes was analyzed by immunohistochemistry .Results Levels of total AMPK protein of cardiomyo-cytes in each group showed no obvious changes .Compared with the control group , AICAR treatment up-regulated the p-AMPK protein level （P〈0.01）.Compared with the AICAR group, Compound C treatment down-regulated the p-AMPK protein level （P〈0.05）.Compared with the control group , the surface area of cardiomyocytes （P〈0.01）, and ANP mR-NA expression （P〈0.01） in the PE group was significantly increased;the 3-MH release in medium of cardiomyocytes in the PE group was significantly decreased （P〈0.01）.The 3-MH release was lower in the PE group as compared with the control group, the 3-MH release of the PA gro

关 键 词：单磷酸腺苷激活蛋白激酶 心肌细胞 肥大 

分 类 号：R364.3[医药卫生—病理学] R541[医药卫生—基础医学]