济生肾气汤加三七、鳖甲对CCl4诱导肝纤维化大鼠模型的干预作用及机制

Intervention effect and mechanism of Jisheng Shenqi Decoction plus Panax notoginseng and Bionjia on CCL4-induced hepatic fibrosis rat model

目的:探讨济生肾气汤加三七、鳖甲对四氯化碳(CCl;)诱导的肝纤维化大鼠模型的干预作用及具体机制。方法:将SPF级SD大鼠随机分为对照组、模型组、中药治疗(低、中、高剂量)组,模型组与中药治疗组采用腹腔注射40%CCl4橄榄油溶液,每周2次,持续8周的方法复制肝纤维化大鼠模型。造模成功后,各给药组分别灌胃相应药物,对照组及模型组予等体积蒸馏水灌胃,每天1次,治疗4周。末次灌胃结束后,对肝组织切片进行HE染色和Masson染色观察肝组织病理形态学,检测肝功能指标(ALT、AST);ELISA法测定转化生长因子β(TGF-β)、血管紧张素Ⅱ(Ang-Ⅱ)、壳多糖酶3样蛋白1(CHI3L1)、白介素-1(IL-1)、白介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)含量;RT-PCR法测定Ⅰ型、Ⅲ型胶原蛋白(Col-Ⅰ、Col-Ⅲ)及TGF-βmRNA的表达水平。结果:HE和Masson染色显示模型组大鼠大量肝细胞炎症坏死,肝组织胶原纤维广泛增生,纤维间隔相互连接,假小叶形成,而中药各剂量治疗组大鼠的肝脏炎症及肝纤维化程度均较模型组减轻;与模型组比较,中药治疗组ALT、AST、TGF-β、Ang-Ⅱ、CHI3L1、IL-1、IL-6、TNF-α水平均下降,Col-Ⅰ、Col-Ⅲ、TGF-βmRNA表达均减少,其中以中药中、高剂量组效果最为明显(P<0.01)。结论:济生肾气汤加三七、鳖甲对CCL4诱导的大鼠肝纤维化具有明显的改善作用,其作用机制可能与下调血管紧张素转换酶(ACE)-血管紧张素Ⅱ(Ang-Ⅱ)-血管紧张素Ⅱ受体1(AT1R)信号通路相关基因表达,降低相关细胞因子水平、促进细胞外基质降解方面等有关。

Objective:To investigate the interventional effect of Jisheng Shenqi Decoction plus Panax notoginseng and biometria on rat model of liver fibrosis induced by carbon tetrachloride(CCl4) and its specific mechanism. Methods: SPF SPRague-dawley rats were randomly divided into control group,model group,Chinese medicine treatment group(low,medium and high dose)group. The model group and Chinese medicine treatment group were intraperitoneally injected 40%CCl4olive oil solution twice a week for 8 weeks to replicate the rat model of liver fibrosis. After modeling,each administration group was intragastrically given corresponding drugs,control group and model group were intragastrically given equal volume of distilled water,once a day,for 4 weeks. After the last intragastric administration,liver tissue sections were stained by HE and Masson to observe the pathological morphology of liver tissue and detect liver function(ALT and AST). The contents of transforming growth factorβ(TGF-β),angiotensin Ⅱ(Ang-Ⅱ),CHI3L1,INTERleukin-1(IL-1),interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)were determined by ELISA. The mRNA expression levels of Col-Ⅰ,Col-Ⅲ and TGF-β were determined by RT-PCR.Results:HE and Masson staining showed a large number of inflammatory necrosis of liver cells,extensive proliferation of collagen fibers,interconnective fibrous septa and formation of false lobules in liver tissue in model group. Compared with the model group,the levels of ALT,AST,TGF-β,Ang-Ⅱ,CHI3L1,IL-1,IL-6 and TNF-α were decreased in the TCM treatment group,while the mRNA expressions of Col-Ⅰ,Col-Ⅲ and TGF-β were decreased in the TCM treatment group,and the effect was most obvious in the medium-dose and high-dose groups(P<0.01).Conclusion:Jishengqi Decoction plus Panax notoginseng and bionjia can significantly improve CCl4-induced hepatic fibrosis in rats,and the mechanism may be related to down-regulation of angiotensin converting enzyme(ACE)-angiotensin Ⅱ(Ang-Ⅱ)-angiotensin Ⅱ receptor 1(AT1R)signaling pathway. Reduce the level of related cytokines and promote the degradation of extracellular matrix.