摘要
目的:观察钙-钙调素依赖性蛋白激酶(CaMK)Ⅱ抑制剂(autocamtide 2-相关抑制肽,AIP)在大鼠心肌成纤维细胞增殖中的作用及其分子机制。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(CON),血管紧张素Ⅱ(AngⅡ,0.1 umol/L)组,AngⅡ(0.1μmol/L)+AIP(0.2μmol/L)组,AngⅡ(0.1μmol/L)+AIP(0.5μmol/L)组,AngⅡ(0.1μmol/L)+AIP组(1.0μmol/L);MTT法及细胞记数法分别测定心肌成纤维细胞增殖;ELISA法测定细胞因子(TGF-β_1,TNF-a);RTPCR检测基质金属蛋白酶-2,9(MMP-2,9)mRNA水平;免疫印记法检测MMP-2,9的蛋白表达。结果:AIP(0.5μmol/L,1.0μmol/L)抑制AngⅡ引起的心肌成纤维细胞增殖,并呈剂量依赖;AIP(0.5μmol/L,1.0μmol/L)抑制AngⅡ引起的细胞因子分泌,并呈剂量依赖AIP(0.5μmol/L,1.0μmol/L)预防0.1μmol/LAngⅡ引起的MMP-2,9 mRNA及蛋白的表达增加。结论:AIP(0.5μmol/L,1.0μmol/L)对AngⅡ诱导的心肌纤维化具有保护作用,其机制可能与AIP预防心肌成纤维细胞增殖、细胞因子分泌以及对基质金属蛋白酶的调节有关。
AIM.To observe the role of calcium calmodulin dependent protein Kinase （CaMK）Ⅱinhibitor（autocamtide 2-related inhibitory peptide,AIP） in rat cardiac fibroblasts proliferation and their molecular mechanism. METHODS：Using cultured cardiac fibroblasts from neonatal 1- 3 days rat（3 generation）; This cells were divided into five groups as follows： control group,AngⅡgroup（0.1μmol/L）,AngⅡ（0.1μmol/L）＋AIP group（0.2μmol/L）,AngⅡ（0.1μmol/L））＋AIP group （0.5μmol/L）,AngⅡ（0.1μmol/L）＋AIP group（1.0μmol/L）;The cardiac fibroblasts proliferation was detected by cell counting and MTT;The cytokines excreting（TGF-β1,TNF-a） was investigated by ELISA.The expression of MMP-2,9 mRNA was detected by RT-PCR. RESULTS：AIP（0.5μmol/L,1.0μmol/L） could prevent cardiac fibroblasts proliferation in- duced by angiotensinⅡin a dose-dependent manner;AIP（0.5μmol/L,1.0μmol/L） could prevent cytokines excreting induced by angiotensinⅡin a dose-dependent manner;AIP（0.5μmol/L,1.0μmol/L） could prevent the increasing expression of MMP-2,9 mRNA and MMP-2, 9 protein expression induced by 0.1μmol/L AngⅡ.CONCLUSION：AIP（0.5μmol/L,1.0μmol/L） could prevent myocardial fibrosis induced by angiotensinⅡ;The probably mechanism was that：AIP could prevent cardiac fibroblasts proliferation,its cytokines excreting and regulate extracellular matrix（MMP-2,9）.
出处
《中国临床药理学与治疗学》
CAS
CSCD
2010年第4期422-428,共7页
Chinese Journal of Clinical Pharmacology and Therapeutics
