密点麻蜥对小鼠肝纤维化的干预作用及机制

Intervention effect and mechanism of Eremias multiocellata on hepatic fibrosis in mice

目的研究密点麻蜥对肝纤维化模型小鼠的干预作用及机制。方法将60只ICR小鼠随机抽取10只为空白组,其余50只腹腔注射四氯化碳(CCl_(4))-玉米油混合液造模8 w,诱导肝纤维化模型。检测模型制备成功后,将小鼠随机分为模型组、安络化纤组和密点麻蜥低、中、高浓度组,持续灌胃4 w。治疗后采用苏木素-伊红(HE)染色法观察小鼠肝组织病理学变化;用酶联免疫吸附试验检测小鼠血清透明质酸(HA)、层黏连蛋白(LN)、Ⅲ型前胶原(PC-Ⅲ)、Ⅳ型胶原(C-Ⅳ)及转化生长因子(TGF)-β1、血小板衍化生长因子(PDGF)-BB水平;用Western印迹检测小鼠肝组织中基质金属蛋白酶(MMP)-13及MMP抑制剂(TIMP)-1蛋白表达量。结果与空白组相比,模型组血清中HA、LN、PC-Ⅲ、C-Ⅳ及TGF-β1、PDGF-BB水平均显著升高(P<0.01);与模型组比较,安络化纤组和密点麻蜥高、中、低浓度组血清中HA、LN、PC-Ⅲ、C-Ⅳ及TGF-β1、PDGF-BB水平均显著降低(P<0.01),各浓度中以密点麻蜥高浓度组效果较好;与空白组比较,模型组肝组织中MMP-13表达显著降低,TIMP-1表达显著升高(P<0.05,P<0.01);与模型组比较,安络化纤组和密点麻蜥高浓度组肝组织中MMP-13表达显著升高(P<0.05),安络化纤组和密点麻蜥高、中、低浓度组肝组织中TIMP-1表达显著降低(P<0.01),各浓度中以密点麻蜥高浓度组效果较好。结论密点麻蜥可以通过调节MMP-13及TIMP-1表达,调控肝细胞外基质(ECM)沉积和降解平衡;降低TGF-β1和PDGF-BB表达水平,而抑制HSC活化,有效改善小鼠肝纤维化。

Objective To study the intervention effect and mechanism of Eremias multiocellata on liver fibrosis model mice.Methods 10 of the 60 ICR mice were randomly selected as blank group,and the other 50 mice were intraperitoneally injected with tetrachloromethane(CCl_(4))-corn oil mixture for 8 w to induce liver fibrosis model.After the model was successfully established,the mice were randomly divided into model group,Anluohuaxian group,low,medium and high concentration groups of Eremias multiocellata and all groups were treated by gavage for 4 w.After treatment,htoxylin-eosin(HE)staining was used to observe the pathological changes of mice liver tissue;hyaluronic acid(HA),laminin(LN),procollagen typeⅢ(PC-Ⅲ),collagen typeⅣ(C-Ⅳ),transforming growth factor(TGF)-β1 and platelet derived growth factor(PDGF)-BB in serum of mice were detected by ELISA.The expressions of matrix metalloproteinase(MMP)-13 and MMP inhibitor(TIMP)-1 in liver tissue were detected by Western blot.Results Compared with the blank group,the levels of serum HA,LN,PC-Ⅲ,C-Ⅳ,TGF-β1 and PDGF-BB in the model group were significantly increased(P<0.01).Compared with the model group,the levels of serum HA,LN,PC-Ⅲ,C-Ⅳ,TGF-β1 and PDGF-BB in Anluohuaxian group and high,medium and low concentration groups of Eremias multiocellata were significantly decreased(P<0.01),and among all the concentrations,the effect of high concentration group was better.Compared with the blank group,the expression of MMP-13 in liver tissue of the model group was significantly decreased,and the expressions of TIMP-1 was significantly increased(P<0.05,P<0.01).Compared with the model group,the expressions of MMP-13 in liver tissue of mice in Anluohuaxian group and high concentration group of Eremias multiocellata were significantly increased(P<0.05),and the expressions of TIMP-1 in the liver tissue of the Anluohuaxian group and the high,medium and low concentration groups of Eremias multiocellata were significantly decreased(P<0.01),and among all the concentrations,the effect of high concentration group was better.Conclusions Eremias multiocellata could regulate the balance of liver extracelluar matrix(ECM)deposition and degradation by regulating the expressions of MMP-13 and TIMP-1,it also could reduce the expression levels of TGF-β1 and PDGF-BB to inhibit HSC activation and effectively improve liver fibrosis in mice.