摘要
目的研究细胞外基质(ECM)的组成和结构变化对基质金属蛋白酶-2(MMP-2)、膜型基质金属蛋白酶-1(MT1-MMP)及基质金属蛋白酶组织抑制物(TIMPs)表达及活性的影响,探讨MMP-2、TIMPs和MT1-MMP对细胞外基质降解的调节作用。方法将人肝星状细胞分别培养于不同的细胞外基质中,采用酶谱分析测定MMP-2;Westernblot测定MT1-MMP的表达量;反向酶谱分析测定TIMPs的表达量;RT-PCR测定MMP-2、TIMPs和MT1-MMP的mRNA表达量。结果I型胶原表面和内部培养的人肝星形细胞(HSC)的MMP-2酶原及MMP-2,TIMP-2,MT1-MMP及其mRNA表达明显增强,而未聚合的I型胶原单体分子则无此作用,类似于基底膜成分的Matrigel对MMP-2的表达也没有明显影响。结论结论在不同的细胞外基质成分中,HSC的MMP-2,TIMPs和MT1-MMP表达量不同;在细胞外基质降解的过程中,MMP-2、TIMPs和MT1-MMP可能一起对细胞外基质的合成和降解进行调节。
OBJECTIVE To study the effects of extracellular matrix ( ECM ) on expression of matrix metalloproteinase-2 ( MMP- 2), membrane type Ⅰ matrix metalloproteinase(MT1-MMP) and tissue inhibitors of matrix metalloproteinases (TIMPs) of HSC, and inquire into the modulation of MMP-2, MT1-MMP and TIMPs on the degradation of ECM. METHODS Human hepatic stellate cells were cultured in different components of extracellular matrix. Employ the methods of zymogram analysis to determine the expression of MMP-2, western-blot to determine MT1-MMP, reverse zymogram analysis to determine the expression of TIMPs, RT-PCR to determine the expression of mRNA of MMP-2, MT1-MMP and TIMPs. RESULTS Human HSCs cultured in the type Ⅰ collagen gel and on its surface,pro-MMP-2 and its mRNA, MMP-2 and its mRNA,TIMP-2 and its mRNA, MT1-MMP and its mRNA are obviously increased, but not in the unpolymerized collagen Ⅰ, as same as the basement membrane components Matrigel. CONCLUSION In different components of ECM, there were different expressions of MMP-2, MT1-MMP and TIMP-2. We assumed that MMP-2, MT1-MMP and TIMP-2 might give the synthesis and degradation of ECM a modulation together .
出处
《中国现代应用药学》
CAS
CSCD
北大核心
2008年第5期389-393,共5页
Chinese Journal of Modern Applied Pharmacy