腺苷A1受体可通过细胞外信号调节激酶通路抑制血管紧张素Ⅱ诱导的心肌细胞肥大  

Stimulation of adenosine A1 receptor attenuates angiotensinⅡ induced myocardial hypertrophy inneonatal rats via the extracellular signal-regulated kinase signal pathways

作　　者：李志业[1] 杨育红[2] 邢玲[1] 

机构地区：[1]郑州大学第二附属医院药学教研室,450001 [2]辽宁医学院药理教研室辽宁省心血管药物重点实验室

出　　处：《中华心血管病杂志》2013年第8期698-703,共6页Chinese Journal of Cardiology

基　　金：辽宁省教育厅团队项目（2009T064）;辽宁省科技厅科技计划项目（2009225010-40）

摘　　要：目的探讨腺苷A1受体激活对血管紧张素Ⅱ（AngⅡ）诱导的心肌细胞肥大的抑制作用及其机制。方法体外培养大鼠乳鼠心肌细胞，以AngⅡ0．1p．mol／L诱导心肌细胞肥大，观察腺苷A1受体激动剂R（-）-N6-（2-phenyllsopropyl）adenosine（R—PIA）1μmo[／L对其的影响，并进一步探讨细胞外信号调节激酶1／2（ERKl／2）特异性抑制剂1，4-Diamino-2，3-dicyano-1，4-bis（0-aminophenylmercapto）butadiene（U0126）1μmol／L、PKC抑制剂Ro·31—822050nmol／L、百日咳毒素（PTX）5mg／L干预后，腺苷A1受体的激活对心肌细胞肥大的影响及其机制。通过Lowry法测心肌细胞蛋白含量，RT—PCR法检测心肌细胞心钠素（ANP）mRNA的表达水平，Westernblot法检测心肌细胞磷酸化细胞外信号调节激酶（p-ERKl／2）及ERKl／2的相对表达水平，以Fluo-3／AM为荧光探针，共聚焦显微镜下测量心肌细胞钙离子瞬变（[Ca_2＋]i瞬变）。结果0．1μmol／LAngⅡ可以诱导心肌细胞肥大（P〈0．01），腺苷A1受体激动R—PIA可以使肥大心肌细胞蛋白含量降低、ANP的mRNA表达减少、ERKl／2相对表达降低、[ca_2＋]i荧光强度减小（P均〈0．05）。U0126、Ro-31—8220有类似抑制作用，且Ro-31—8220可以部分阻断R-PIA对心肌肥大细胞的抑制作用。PTX预处理后，R-PIA对Ang11诱导的心肌细胞肥大的抑制作用消失。结论腺苷A1受体可以通过ERKl／2通路抑制AngII诱导的心肌细胞肥大，其机制与降低细胞内[ca_2＋]i浓度及p-ERKl／2表达有关。Objective To observe the impact of adenosine A1 receptor stimulation on extracellular signal-regulated kinasel/2 （ERK1/2） signal pathways on angiotensin Ⅱ （Ang ]I ） stimulated cardiomyocytes of neonatal rats in vitro. Methods Cardiomyocytes of neonatal rats were cultured in vitro. Cardiomyocytes hypertrophy was induced by Ang Ⅱ （0. 1 μmol/L ）. The antihypertrophic effect of adenosine A1 receptor stimulation via adenosine A1 receptor agonist R-PIA （ 1 μmol/L） was observed in the presence or absence of ERK1/2 inhibitor 1,4-Diamino-2,3-dicyano-1,4-bis（o-aminophenylmercapto） butadiene （U0126） 1 μmol/L, PKC inhibitor Ro-31-8220 （50 nmol/L）, and pertussistoxin （PTX, 5 mg/L）. The total protein content was assayed by the method of Lowry. The expression of mRNA of atrial natriuretic peptide（ ANP）was determined by RT-PCR. [ Ca2＋ ] i was measured by eonfocal microscope using Fluo-3/AM as flouresecent indicator. The relative expression of ERK1/2 was determined by Western blot. Results Compared with normal control group, AngⅡ induced significant cardiomyocyte hypertrophy. Compared with AngⅡ group, R-PIA significantly inhibited Ang Ⅱ -induced increase of the protein content, cardiomyocytes volume and expression of ERK1/2, calciumion fluorescence intensity, similar as U0126 and Ro-31-8220. The inhibitory effects onAng Ⅱ induced cardiomyocytes hypertrophy of R-PIA were lost when preincubated with PTX. Conclusion Adenosine A1 receptor can inhibit Ang Ⅱ induced cardiomyocyte hypertrophy through downregulating ERK signal pathways and reducing intracellular Ca2＋

关 键 词：心肌病 肥厚性 腺苷 血管紧张素Ⅱ 细胞外信号调节MAP激酶类 

分 类 号：R542.2[医药卫生—心血管疾病]