Autocamtide 2-相关抑制肽对心肌成纤维细胞分泌细胞因子及基质金属蛋白酶表达的抑制研究  

Study of autocamtide 2-related inhibitory peptide prevent cardiac fibroblasts excreting cytokines and expressing matrix metalloproteinases

作　　者：钟拥军[1] 陈东芹[1] 姚小燕[2] 

机构地区：[1]浙江省嘉兴市第一医院药剂科 [2]浙江省嘉兴市第一医院心内科

出　　处：《中国临床药理学与治疗学》2010年第4期422-428,共7页Chinese Journal of Clinical Pharmacology and Therapeutics

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）.

关 键 词：心肌成纤维细胞 血管紧张素Ⅱ 钙-钙调素依赖性蛋白激酶Ⅱ autocamtide 2-相关抑制肽 基质金属蛋白酶-2 9 

分 类 号：R542.2[医药卫生—心血管疾病] R969[医药卫生—内科学]