摘要
目的:观察钙-钙调素依赖性蛋白激酶(CaMK)Ⅱ在血管紧张素Ⅱ(AngⅡ)或电场刺激(EFS)诱导的大鼠心肌成纤维细胞增殖、分泌细胞因子及胶原酶表达中的作用及其机制。方法:培养新生1-3 d乳鼠心肌成纤维细胞(3代),分为正常对照组(control)、0.1μmol/L AngⅡ组、0.1μmol/L AngⅡ+0.5μmol/L CaMKⅡ抑制剂KN92组、0.1μmol/L AngⅡ+0.5μmol/L CaMKⅡ抑制剂KN93组、0.1μmol/L AngⅡ+0.5μmol/L CaMKⅡ抑制剂AIP组;10 V1.0 Hz EFS组、10 V 1.0 Hz EFS+0.5μmol/L KN92组、10 V 1.0 Hz EFS+0.5μmol/L KN93组、10 V1.0 Hz EFS+0.5μmol/L AIP组、10 V1.0 Hz EFS+0.1μmol/L AngⅡ组。MTT法测定心肌成纤维细胞增殖;ELISA法测定细胞因子(TGF-β1,TNF-α)分泌;RT-PCR检测TGF-β1、TNF-α、collagenⅠ、ⅢmRNA水平。结果:CaMKⅡ抑制剂(0.5μmol/L KN93,0.5μmol/L AIP)能预防AngⅡ或EFS诱导的心肌成纤维细胞增殖;CaMKⅡ抑制剂(0.5μmol/L KN93,0.5μmol/L AIP)可预防AngⅡ或EFS引起的细胞培养上清液TGF-β1、TNF-α含量及相应mRNA表达增加。CaMKⅡ抑制剂(0.5μmol/L AIP,1.0μmol/L AIP)预防0.1μmol/L AngⅡ引起的collagenⅠ、Ⅲ表达增加。结论:抑制CaMKⅡ对AngⅡ或EFS诱导的心肌成纤维细胞增殖具有预防作用,其机制可能与CaMKⅡ抑制剂抑制TGF-β1、TNF-α以及胶原的表达有关。
AIM： To observe the role of calcium/calmodulin-dependent protein kinase Ⅱ（CaMKⅡ） in the proliferation,released cytokines and expression of collagen Ⅰ and Ⅲ in rat cardiac fibroblasts induced by angiotensin Ⅱ（AngⅡ） or electrical field stimulation（EFS）.METHODS： The cultured cardiac fibroblasts were isolated from the neonatal rats of 1-3 days and used in the 3rd passage.The cells were divided into 10 groups： control group,0.1 μmol/L AngⅡ group,0.1 μmol/L AngⅡ＋0.5 μmol/L KN92 group,0.1 μmol/L AngⅡ＋0.5 μmol/L KN93 group,0.1 μmol/L AngⅡ＋0.5 μmol/L AIP group;10V 1.0 Hz EFS group,10 V 1.0 Hz EFS＋0.5 μmol/L KN92 group,10 V 1.0 Hz EFS＋0.5 μmol/L KN93 group,10 V 1.0 Hz EFS＋0.5 μmol/L AIP group,10 V 1.0 Hz EFS＋0.1 μmol/L AngⅡ group.MTT was used to detect the proliferation of cardiac fibroblasts.The release of cytokines was measured by ELISA.The mRNA expression of TNF-α,TGF-β1 and collagen Ⅰ,Ⅲ was determined by RT-PCR.RESULTS： CaMKⅡ inhibitors（0.5 μmol/L KN93 or 0.5 μmol/L AIP） prevented the proliferation and the increase in the expression of TGF-β1 and TNF-α in cardiac fibroblasts induced by AngⅡ（0.1 μmol/L） or EFS（10 V 1.0 Hz）.CaMKⅡ inhibitors（0.5 μmol/L AIP or 1.0 μmol/L AIP） also prevented the increase in mRNA expression of collagen Ⅰ and Ⅲ induced by 0.1 μmol/L AngⅡ.CONCLUSION： Inhibition of CaMKⅡ prevents the proliferation of cardiac fibroblasts induced by AngⅡ or EFS.The possible mechanism of CaMKⅡ inhibitors may be involved in preventing the mRNA expression and release of cytokines（TGF-β1 and TNF-α）,and regulating collagen I and III expression.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2010年第8期1549-1554,共6页
Chinese Journal of Pathophysiology
