罗格列酮对大鼠肝星状细胞基质金属蛋白酶-2表达的影响

Effects of rosiglitazone on matrix metalloproteinase-2 in rats hepatic stellate cells

目的研究罗格列酮激活过氧化物酶体增殖物激活受体γ(PPARγ)后对大鼠肝星状细胞(HSC)表达基质金属蛋白酶-2(MMP-2)的影响,探讨罗格列酮对肝纤维化的影响。方法采用胶原酶原位循环灌流法分离大鼠HSC,MTT法检测不同浓度罗格列酮对HSC增殖的影响,采用逆转录聚合酶链反应(RT-PCR)、酶联免疫吸附试验(ELISA)及酶谱法检测罗格列酮对MMP-2基因、蛋白及酶活性的影响。结果罗格列酮在0～40μmol/L浓度范围内抑制HSC增殖。5μmol/L、10μmol/L罗格列酮对HSCMMP-2mRNA表达无明显影响。与对照组相比,5μmol/L、10μmol/L罗格列酮通过激活PPARγ抑制MMP-2蛋白分泌,使上清中MMP-2的酶活性呈剂量依赖性下降,10μmol/L罗格列酮组可使HSC内MMP-2的酶活性下降。结论罗格列酮可抑制HSCMMP-2蛋白分泌及酶活性,这可能是其抗肝纤维化的机制之一。

Objective To observe the effects of rosiglitazone, synthetic peroxisome proliferator-activated receptor- γ, PPAR-γ agonists on marix metalloproteinase-2 （MMP2） in rats hepatic stellate cells （HSCs）, and explore the underlying mechanisms of PPAR-γ in hepatic fibrogenesis. Methods HSCs were isolated from normal rat liver tissue and the first passage cells were conducted in all experiments. HSCs proliferation was detected by MTT colormetric assay. MMP2 mRNA level was detected by RT-PCR and analyzed semi-quantitatively. ELISA method was used to measure MMP2 proteins secreted from HSCs. Zymography was performed to investigate MMP2 enzymatic activities. Results Rosiglitazone inhibited HSC proliferation within the concentration of 0 to 40 umol/L. PPAR-γ gene expression was up-regulated on the treatment of rosiglitazone. Rosiglitazone did not affect MMP2 mRNA expression. Compared with control group, 5 umol/L and 10umol/L rosiglitazone could significantly decrease MMP2 protein secretion and dose-dependently downregulate the enzymatic activities of MMP2 in the culture medium, moreover, enzymatic activities of MMP2 in HSCs was inhibited by 10 umol/L rosiglitazone. Conclusion Rosiglitazone could inhibit protein excretion and enzymatic activities of MMP2, which might be one of its antifibrotic mechanisms.