复方861对DMN损伤性大鼠肝纤维化肝脏中MMP-2表达的影响

Effect of Chinese herbs compound 861 on the expression of MMP-2 in the DMN injured fibrotic liver in rats

目的研究抗肝纤维化中药复方861对DMN诱导的肝纤维化大鼠肝脏中MMP-2表达的影响。方法将大鼠随机分成正常对照组,模型组与861防治组。模型组以DMN腹腔内注射,第1周注射3次,随后每周2次,共注射6周;治疗组同法注射DMN同时每日灌胃861冲剂;正常对照组以生理盐水代替腹腔内注射。实验结束处死大鼠,留取的肝脏组织做HE与Masson染色按0～4期标准判定肝纤维化程度,应用半定量RT-PCR检测肝组织中MMP-2mRNA,用免疫组化检测肝组织中MMP-2的表达。结果模型组大鼠肝纤维化程度处于2～4期,其中大部分达3期以上,861防治组大鼠肝纤维化程度明显减轻,只有少部分达3期。模型组大鼠肝组织中MMP-2mRNA水平显著高于正常对照组(t=4.03,P<0.01),而861防治组大鼠肝组织中MMP-2mRNA水平显著低于模型组(t=3.43,P<0.01)。861防治组与正常组之间,肝组织中MMP-2mRNA水平无明显差别。模型组大鼠肝组织MMP-2的阳性表达显著强于正常对照组(t=4.56,P<0.01),861防治组肝组织中MMP-2的阳性表达虽明显强于正常组(t=2.71,P<0.05),但较模型组显著减弱(t=3.24,P<0.01)。结论中药复方861能有效抑制DMN诱导的肝纤维化大鼠肝脏中MMP-2的表达,可能有利于维持HSC处于静止状态所需要的细胞外基质环境,这对于抑制HSC的活化与肝纤维化的发展具有重要意义。

Objective To explore the effect of Chinese herbs compound 861 on the expression of MMP-2 in the DMN injured fibrotic liver in rats. Methods Wistar rats were randomly alocated into normal group, model group and Chinese compound 861 （Cpd 861） group. To induce liver fibrogenesis, rats were intraperitoneally injected with dimethylnitrosamine （DMN） three consecutive times in the first week, then two consecutive times per week, totally for 6 weeks. In normal control group, rats were treated with saline by the same method as model group. In the Cpd 861 group, rats were fed with Cpd 861 （3g/kg BW）while treated with DMN until the end of study period. Animals weresacrificed at the end of the experiment. Histological examinations were performed after hematoxylin and eosin staining, and Masson staining, the fibrosis stage was classified into 0 to 4. The hepatic mRNA level of MMP-2 was determined by semi-quantitive RT-PCR, histological expression of MMP-2 was detected by immunohistochemistry method. Results The rat model was successfully induced by intraperitoneally injection with DMN. In model group, the extent of liver fibrosis among most animals were up to more than 3th stage. In Cpd 861 group, the extent of liver fibrosis were obviously degraded than the model group. In model group, the hepatic MMP-2 mRNA level was significantly higher than the normal group（t=4.03 ,P〈0.01）. In Cpd 861 group, the hepatic MMP-2 mRNA level was significantly lower than the model group. The hepatic MMP-2 mRNA levels between the Cpd 861 group and the normal group were not obviously different. In model group, the positive expression of MMP-2 in liver tissue was significantly stronger than the normal group（t=4.56 ,P〈0.01）. In Cpd 861 group, the positive expression of MMP-2 in liver tissue was obviously stronger than the normal group （t=2.71 ,P〈0.05）, but was significantly weaker than the model group （t=3.24,P〈0.01）. Conclusion Chinese herbs compound 861 can effectively inhibit the MMP-2 expression in the fibrotic liver induced by DMN in rats,that may be propitious to prevent the basement membrane like extracellular matrix from degradation by MMP-2, and be in favor of keeping HSCs quiescent. It means the great sense that keep back HSC＇s activation and hold back the development of liver fibrosis.