摘要
目的探讨钙-钙调素依赖性蛋白激酶(CaMK)Ⅱ对大鼠心肌成纤维细胞增殖的影响及其机制。方法培养新生SD大鼠心肌成纤维细胞,分为正常对照组,AngⅡ(0.1μmol/L)组。AngⅡ+AIP(0.2μmol/L)组,AngⅡ+AIP(0.5μmol/L)组,AngⅡ+AIP(1.0μmol/L)组;EFS(10V1.0Hz)组,EFS+AIP(0.2μmol/L)组,EFS+AIP(0.5μmol/L)组,EFS+AIP(1.0μmol/L)组;MTT法及细胞记数法分别测定心肌成纤维细胞增殖;免疫印记法检测CaMKⅡδB的蛋白表达;RT-PCR检测CaMKⅡδB、C mRNA水平。结果 CaMKⅡ抑制剂能抑制AngⅡ或EFS诱导的心肌成纤维细胞增殖(P<0.05);抑制AngⅡ引起的CaMKⅡδB、δC表达增加(P<0.05);AngⅡ刺激24h CaMKⅡ活性升高(P<0.05),48h升高更明显(P<0.01)。当电压从10V增加至40V,CaMKⅡ活性不断增加,呈电压依赖(P<0.05)。结论 CaMKⅡ阻断剂对心肌成纤维细胞增殖有抑制作用,可能与抑制CaMKⅡδB和δC表达有关。
Objective To study the effects of calcium calmodulin dependent protein Kinase （CaMK） Ⅱon cardiac fi-broblast proliferation of rat and its mechanisms. Methods Cardiac fibroblasts cultured from neonatal SD rat were divid-ed into nine groups as follows：control group, Ang II （0.1μmol/L）group,Ang II＋AIP （0.2μmol/L）group, Ang II＋AIP （0.5μmol / L）group, AngⅡ＋ AIP（1.0μmol / L）group, 10V1.0Hz EFS group, EFS ＋0.2 μmol / L AIP group, EFS ＋0.5μmol/L AIP group, EFS＋1.0μmol/L AIP group. Cel counting and MTT were performed to detect cardiac fibroblasts proliferation. Protein expression of CaMKII-δB was measured by Western blot. mRNA expression of CaMK II- δB and CaMK II- δC was determined by RT-PCR. Results CaMKII inhibitor prevented cardiac fibroblasts proliferation in-duced by AngⅡor EFS（P〈0.05）and inhibited expression of CaMKⅡδB and δC induced by Ang II（P〈0.05）. The ac-tivity of CaMKII increased when being exposed to Ang II for 24 hours（P〈0.05）and more significantly for 48h（P〈0.01）. EFS enhanced CaMKII activation in a voltage-dependent manner with the voltage from 10V to 40V（P〈0.05）. Conclusion CaMKII inhibitor can prevent cardiac fibroblasts proliferation by inhibiting expression of CaMKⅡδB andδC.
出处
《心电与循环》
2013年第3期188-192,共5页
Journal of Electrocardiology and Circulation
