CCl4肝纤维化过程中肝组织金属硫蛋白与肝星状细胞MMP-2表达及活性之间相关性研究

目的： 研究金属硫蛋白（MT）在体内及体外与基质金属蛋白酶-2(MMP-2)表达、活性之间的相关性。方法: 昆明种雄性成年小鼠50只，其中实验鼠 40只，对照鼠10只，用CCl4诱导小鼠肝纤维化模型。向肝星状细胞（HSCs）条件培养基（含有MMP-2）中加不同浓度MT，使之与MMP-2酶学共浴，检测MMP-2活性。制作小鼠肝组织芯片，天狼猩红染色判断肝纤维化程度。免疫组化法测肝组织MMP-2、MT蛋白表达，酶图法测鼠肝组织匀浆和培养基中MMP-2活性。结果: ①模型组小鼠肝组织MMP-2和MT蛋白表达水平均随纤维化进展而呈波浪式交替升高，但二者时相变化相反。 ②模型组小鼠肝组织MMP-2活性随实验时间的增加而逐渐升高，亦有波动。除个别时点外，酶活性与MT表达呈负相关。③肝星状细胞条件培养基与不同浓度的MT直接作用后，MMP-2活性随MT浓度增加活性逐渐降低(P＜0.01)。结论: 在小鼠肝纤维化发生发展过程中，肝组织MT蛋白和MMP-2蛋白表达均有上升趋势，二者间可能存在相互作用；MT蛋白和MMP-2活性呈负相关，前者对后者可能起抑制作用。

Correlation between metallothionein and matrix metalloproteinase-2 expression and activity in rats during CC14-induced hepatic fibrogenesis

AIM: To investigate the relevance between metallothionein(MT) and matrix metalloproteinase-2 (MMP-2) expression and activity in hepatic stellate cells (HSCs). METHODS: Fifty Kunming male mice, weighing 25 g±5 g, were randomly divided into two groups: experiment group (40 mice) and control group (10 mice). CCl4 was used to induce the hepatic fibrosis model in the experiment group. Conditioned medium of HSCs (containing MMP-2) was added at different concentrations of MT, then MMP-2 activity was detected. Liver tissue microarray was used. Collagen fibers was detected by Sirius red staining. The expressions of MMP-2 and MT proteins in liver tissues were detected by immunohistochemistry staining. Gel zymography was used to confirm the activity of MMP-2 in liver tissue homogenate and in the conditioned medium. RESULTS: The expressions of both MMP-2 and MT proteins in liver tissues increased fluctuating with the process of fibrogenesis in the model mice alternately, but the proteins showed out of phase changes. The activity of MMP-2 in the liver tissues also increased gradually and fluctuated with the development of fibrosis. Apart from individual time points, the activity of MMP-2 and MT expression were negatively correlated. The activity of MMP-2 in the conditioned medium treated by MT declined in a dose-dependent manner (r=-0.9990, P<0.01). CONCLUSION: The results suggest that there is an upward tendency in the expression of MMP-2 and MT proteins in the liver fibrogenesis, but interaction between them may be exist, and MT inhibits the activity of MMP-2.