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在体外培养条件下观察了凝血酶对人肾小球系膜细胞表达细胞外基质降解酶——明胶酶A(MMP-2)及明胶酶B(MMP-9)的调节作用。发现在无血清RPMI1640培养12h后,酶谱法显示系膜细胞培养上清液中MMP-2活性极低,未检测出明显的MMP-9酶解活性,但加入凝血酶后,可剂量依赖性地上调其MMP-2、MMP-9活性;RT-PCR结果表明,在无血清RPMI1640培养12h后,系膜细胞仅表达低水平MMP-9mRNA,加入凝血酶后,MMP-9mRNA转录水平随刺激浓度的增加而明显上调,但凝血酶对MMP-2mRNA的转录过程无明显影响;凝血酶的上述作用可被其特异性阻断剂——水蛭素部分性地拮抗。说明凝血酶分别在基因转录和(或)蛋白质修饰水平调节人肾小球系膜细胞表达MMP-2、MMP-9,从而参与肾小球组织重构及炎症性损伤。
The effect of thrombin on the expression of extracellular matrix-degrading enzyme-MMP-2 and MMP-9 in human glomerular mesangial cells was observed under in vitro culture conditions. It was found that after cultured in serum-free RPMI1640 for 12h, the activity of MMP-2 in mesangial cell culture supernatant was very low, and no obvious activity of MMP-9 was detected. However, the addition of thrombin showed dose-dependent The results of RT-PCR showed that the mesangial cells only expressed low levels of MMP-9 mRNA after 12 h of culture in serum-free RPMI1640. After addition of thrombin, the transcription level of MMP-9 increased with the stimulation concentration But thrombin had no significant effect on the transcription of MMP-2mRNA. The above-mentioned effect of thrombin could be partially antagonized by its specific inhibitor hirudin. It is concluded that thrombin regulates the expression of MMP-2 and MMP-9 in human glomerular mesangial cells by gene transcription and / or protein modification, respectively, and thus participates in glomerular remodeling and inflammatory injury.