咪达普利拉对人心脏成纤维细胞基质金属蛋白酶-2和抑制剂-2的作用及机制研究  

Effects of imidaprilat on matrix metalloproteinase-2 and TIMP-2 in human cardiac fibroblasts

作　　者：郭晓纲[1] 陈君柱[1] 朱建华[1] 张芙荣[1] 邱原刚[1] 尚云鹏[1] 

机构地区：[1]浙江大学医学院附属第一医院心内科,浙江杭州310003

出　　处：《中国病理生理杂志》2008年第7期1263-1269,共7页Chinese Journal of Pathophysiology

基　　金：浙江省医药卫生科学研究基金资助项目(No.2007B052);浙江省科技厅钱江人才计划资助项目(No.2006R10024)

摘　　要：目的:研究血管紧张素转化酶抑制剂(angiotensin converting enzyme inhibitor,ACEI)咪达普利活性代谢物咪达普利拉对白细胞介素-1β(interleukin-1β,IL-1β)诱导的心脏成纤维细胞基质金属蛋白酶(matrixmetalloproteinases,MMPs)和基质金属蛋白酶2型抑制剂(type2tissue inhibitor of matrix metalloproteinase,TIMP-2)表达的影响及其可能机制。方法:原代人心脏成纤维细胞从美国细胞应用所购买。用RT-PCR检测心脏成纤维细胞MMP-2、MMP-9和TIMP-2mRNA水平的变化;用凝胶酶谱法分析心脏成纤维细胞中MMP-2、MMP-9的活性;用Griess方法检测培养上清一氧化氮(nitric oxide,NO)水平。结果:通过凝胶酶谱分析,RT-PCR和Griess方法发现IL-1β能显著增加MMP-2基因转录以及活性(P<0.05),同时也显著增加了细胞培养上清中NO的产生(P<0.05)。IL-1β的这些效应可以被咪达普利拉和外源性NO合酶抑制剂L-单甲基精氨酸(NG-methyl L-Arginine,L-NMMA)所抑制(P<0.05),而外源性NO供体亚硝基铁氰化钠(sodium nitroprusside,SNP)可以取消咪达普利拉对于NO、MMP-2的有效作用(P<0.05)。实验中发现IL-1β以及咪达普利拉对TIMP-2基因转录无明显作用(P>0.05)。结论:咪达普利拉能通过NO途径抑制IL-1β诱导的心脏成纤维细胞MMP-2的合成和活性;提示ACEI抗心肌重塑以及心力衰竭的部分作用可能是通过MMPs途径实现的。AIM： To determine the effects and possible signaling pathway of imidaprilat, an active type of angiotensin converting enzyme inhibitor （ ACEI）, on matrix metalloproteinases （MMPs） and type 2 tissue inhibitor of matrix metalloproteinase （ TIMP - 2 ） in human cardiac fibroblasts in the presence of interleukin - 1β （ IL - 1β ）. METHODS ： The primary human cardiac fibroblasts were purchased at passage 2 from Cell Systems. In the experiment, RT - PCR was used for analysis of transcription of MMPs and TIMP - 2. Determination of proteolytic activity was performed with zymogra- phy. Assessment of nitric oxide （NO） production in the culture medium was performed by Griess reagent. RESULTS： IL - 1 β significantly increased MMP - 2 activity, transcription and NO production in the supernatant of culture medium. These effects of IL - 1β were inhibited by imidaprilat or NO synthase inhibitor NG - methyl L - arginine. Sodium nitroprusside, an exogenous NO donor, prevented significantly the effects of imidaprilat on MMP - 2 inhibition. Imidaprilat alone didn＇t affect MMP -2 activity and expression. Neither IL - 1 β nor imidaprilat had effect on TIMP -2 transcription in cardiac fibroblasts. CONCLUSION： Imidaprilat inhibits MMP -2 activity and expression in human cardiac fibroblasts induced by IL - 1β via NO - dependent pathway. These data suggest that the beneficial effect of ACEI against left cardiac remodeling and heart failure may be due at least in part to regulating MMPs activity and expression by modulation of NO pathway.

关 键 词：心脏 成纤维细胞 白细胞介素1 一氧化氮 基质金属蛋白酶 血管紧张素转换酶抑制剂 

分 类 号：R363[医药卫生—病理学]