CCl_4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  

Correlation between metallothionein and matrix metalloproteinase-2 expression and activity in rats during CCl_4-induced hepatic fibrogenesis

作　　者：刘蕾蕾[1] 陈平圣[1] 刘东风[1] 

机构地区：[1]东南大学基础医学院病理教研室,江苏南京210009

出　　处：《中国病理生理杂志》2009年第2期351-355,共5页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.30470780)

摘　　要：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。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.

关 键 词：肝硬化 金属硫蛋白 基质金属蛋白酶-2 小鼠 

分 类 号：R363[医药卫生—病理学]