柔肝化纤颗粒对四氯化碳诱导的肝纤维化大鼠基质金属蛋白酶1及其抑制因子的影响

目的 探讨柔肝化纤颗粒对四氯化碳诱导的肝纤维化大鼠基质金属蛋白酶1（MMP-1）和金属蛋白酶组织抑制因子1（TIMP-1）的影响.方法 建立四氯化碳诱导的肝纤维化大鼠模型,健康清洁SPF级大鼠85只,雌雄各半,随机数字表法分为正常对照组（15只）、病理模型组（16只）、柔肝化纤颗粒组（16只）、大黄座虫丸组（16只）及秋水仙碱对照组（16只）.正常对照组不做其他处理,常规饲养;肝纤维化造模成功后,正常对照组、病理模型组4周后灌胃0.9％氯化钠注射液,柔肝化纤颗粒组、大黄廑虫丸组、秋水仙碱对照组造模成功4周后分别给予柔肝化纤颗粒、大黄=虫丸（0.2 g/kg）、秋水仙碱（100 μg/kg）灌胃,各组灌胃液体量为10 ml/kg体重,每日1次.各组于用药治疗后5、10周,免疫组织化学染色及计算机图像分析技术检测Ⅳ胶原.利用半定量反转录-聚合酶链反应检测MMP-1和TIMP-1 mRNA的表达.结果 用药治疗后5、10周,病理模型组肝组织炎症活动度计分、肝纤维化程度计分均明显高于正常对照组[用药后5周：（4.93±2.56）分比（1.08 ±0.29）分,（15.57±6.12）分比0分,用药后10周：（5.03±2.66）分比（1.10±0.22）分,（16.27 ±6.21）分比0分],差异均有统计学意义（均P＜0.05）;经柔肝化纤颗粒和秋水仙碱、大黄廑虫丸治疗后,大鼠肝组织炎症活动度、肝纤维化程度计分及Ⅳ胶原蛋白水平均明显降低（P＜0.05）.用药治疗后10周,病理模型组MMP-1 mRNA和TIMP-1 mRNA表达分别为（1.30±0.15）和（20.62 ±4.56）,均明显高于正常对照组[分别为（0.94±0.44）、（10.52±3.20）],差异均有统计学意义（均P ＜0.05）;柔肝化纤颗粒组MMP-1 mRNA表达为（1.58±0.34）,明显高于病理模型组、大黄廑虫丸组（1.39±0.35）和秋水仙碱对照组（1.41±0.41）;柔肝化纤颗粒组TIMP-1 mRNA为（13.6±3.3）,明显低于病理模型组、大黄廑虫丸组（18.3±4.4）和秋水仙碱对照组（17.3±4.47）（P＜0.05）;柔肝化纤颗粒组TIMP-1/MMP-1 mRNA为（8.9±2.0）,明显低于病理模型组、大黄廑虫丸组（11.1±1.9）和秋水仙碱对照组（10.4±2.4）（P＜0.05）.结论 柔肝化纤颗粒能通过调节TIMP-1/MMP-1比例来促进细胞外基质降解,从而发挥对肝纤维化的防治作用.

Effects of Rouganhuaxian grannules on matrix metalloproteinase 1 and matrix metalloproteinase 1 inhibitors in rats model of liver fibrasis induced by CCl4

Objective To investigate the effects of Rouganhuaxian grannules on matrix metalloproteinase 1 （MMP-1） and MMP-1 inhibitors （TIMP-1） in rats model of liver fibrasis induced by CCl4.Methods Liver fibrosis model was set up by CCl4,85 rats of health and sweeps were randomly divided into control group,pathologic model group,rouganhuaxian grannules group,Dahuang Zhechong pill group and autumn waters fairy alkali matched control group.The change of MMP-1 and TIMP-4 were check by immunol histochemical method and computer image-analysis technology.Results The severity of inflammation,liver fibrosis scorings and the level of collagen type Ⅳ in pathologic model group were （5.03 ± 2.66）,（16.27 ± 6.21） and （194 ± 15）,and all obviously higher than that in rats of control group （P ＜ 0.05）.Compared with before treatment,the severity of inflammation,liver fibrosis scorings and the level of collagen type Ⅳ were obviously lower after treat for rouganhuaxian grannules group,Dahuang Zhechong pill group and autumn waters fairy alkali matched（P ＜ 0.05）.The level of MMP1,T1MP-1 in pathologic model group were （1.30 ± 0.15） and （20.62 ± 4.56）,and were all obviously higher than those in rats of control group [（0.94 ± 0.44）,（10.52 ± 3.20）] （P ＜ 0.05）.The level of MMP-1 in Rouganhuaxian grannules group was （1.58 ± 0.34）,it was obviously higher than that in pathologic model group,Dahuang Zhechong pill group （1.39 ± 0.35） and autumn waters fairy alkali matched control group（1.41 ± 0.41） （P ＜ 0.05）.But the level of MMP-1 in Rouganhuaxian grannules group was （13.6 ± 3.3）,it was obviously lower than that in pathologic model group,Dahuang Zhechong pill group （18.3 ± 4.4） and autumn waters fairy alkali matched control group （17.3 ± 5） （P ＜ 0.05）.The level of MMP-1 in Rouganhuaxian grannules group was （8.9 ± 2.0）,it was obviously lower than that in pathologic model group,Dahuang Zhechong pill group （11.1 ± 1.9）and autumn waters fairy alkali matched control group（10.41 ±2.37） （P＜0.05）.Conclusion Rouganhuaxian grannules can prevent and cure liver fibrasis through regulating the ratio of MMP-1/TIMP-1.