软肝化坚颗粒对肝纤维化C57小鼠肝组织中MMP-13和TIMP-1 mRNA表达的影响

目的观察软肝化坚颗粒对肝纤维化模型C57小鼠血清中基质金属蛋白酶13(MMP-13)和基质金属蛋白酶抑制剂1(TIMP-1)的含量和肝组织中MMP-13和TIMP-1 mRNA表达的影响。方法 40只C57小鼠被随机分为4组,每组10只,即正常对照组、模型对照组、模型软肝化坚颗粒组和模型秋水仙碱组。采用四氯化碳腹腔注射制造小鼠肝纤维化模型。分别采用ELISA法和实时荧光定量PCR检测小鼠血清MMPs、TIMP-1含量及肝组织中MMP-13和TIMP-1 mRNA的表达。同一指标的多组间计量资料的比较采用方差分析。结果与正常对照组相比较,模型对照组血清中MMPs的含量明显降低,TIMP-1的含量明显升高,P=0.003、0.027,模型软肝化坚颗粒组与正常对照组相比差异均无统计学意义,P=0.364、0.217。与正常对照组相比,模型软肝化坚颗粒组小鼠肝组织中MMP-13 mRNA的表达量显著升高,P=0.005,TIMP-1 mRNA的表达量差异无统计学意义,P=0.997;模型对照组小鼠肝组织中TIMP-1 mRNA的表达量明显升高,差异具有统计学意义,P=0.009。结论软肝化坚颗粒不但可促进MMPs的产生,而且可抑制TIMP-1产生,进而促进ECM的降解,这可能也是软肝化坚颗粒抗肝纤维化的重要机制之一。

Effects of Ruangan Huajian granules on mRNA expression of MMP - 13 and TIMP - 1 in liver tissue among C57 mice with hepatic fibrosis

Objective To observe the effects of Ruangan Huajian granules on the serum levels of matrix metalloproteinase - 13 （ MMP - 13 ） and tissue inhibitor of metalloproteinase - 1 （TIMP - 1） and the mRNA expression of MMP - 13 and TIMP - 1 in liver tissue among C57 mice with hepatic fibrosis. Methods Forty C57 mice were randomly divided into normal control group （n = 10）, model control group （n = 10）, Ruangan Huajian granule group （n = 10）, and colchicine group （n = 10）. A rat model of hepatic fibrosis was established by intraperitoneal injection of carbon tetrachloride in the model control group, Ruangan Huajian granule group, and colchieine group. The serum levels of MMPs and TIMP - 1 and the mRNA expression of MMP - 13 and TIMP - 1 in liver tissue were measured by enzyme - linked immtmosorbent assay and real - time quantitative PCR, respectively. Comparison of continuous data between groups was made by analysis of variance. Results Compared with the normal control group, the model control group had a significantly decreased serum level of MMPs （P =0.003） and a significantly increased serum level of TIMP - 1 （P =0. 027）. There were no significant differences in serum levels of MMPs and TIMP -1 between the normal control group and Ruangan Huajian granule group （P =0. 364 and 0. 217）. Compared with the normal control group, the Ruangan Huajian granule group had significantly increased mRNA expression of MMP - 13 in liver tissue （P =0.005）, but the mRNA expression of TIMP - 1 showed no significant difference between the two groups （P =0.997）. The model control group had significantly increased mRNA expression of TIMP- 1 in liver tissue compared with the normal control group （P = 0.009）. Conclusion Ruangan Huajian granules can not only promote the production of MMPs, but also inhibit the production of TIMP- 1, thus promoting extracellular matrix degradation, which may be one of the action mechanisms of Ruangan Huajian granules in the treatment of hepatic fibrosis.