复肝解毒方对酒精性肝病大鼠肝组织过氧化物酶体增殖物激活受体-γ的激活作用

Activating Effect of Fuganjiedu Formula on Peroxisome Proliferator-Activated Receptor-γ in the Hepatic Tissue in the Rat with Alcoholic Liver Disease

目的 从分子水平探讨中药复方复肝解毒方治疗酒精性肝病的作用机制。方法 选用 4 0只Wister大鼠随机分为正常对照组 (10只 )、模型组 (30只 ) ,16周末断头处死正常对照组 (10只 )和模型组部分大鼠 (9只 ) ,取血清和肝组织标本。模型组另 2 0只随机分为中药治疗组和治疗对照组 ,每组 10只大鼠 ,2 4周后一并处死。分别以HE、SudanⅣ、Masson三色染色观察各组大鼠肝组织光镜下的病理改变和电镜下超微结构的变化。以免疫组织化学染色和RT PCR观察复肝解毒方对过氧化物酶体增殖物激活受体 (PPARγ)蛋白和基因表达的影响。结果 模型组大鼠血清丙氨酸转移酶(ALT)、肿瘤坏死因子 (TNFα)水平和肝匀浆丙二醛 (MDA)含量增高 ,肝组织表现有明显的脂肪变性、炎症、坏死和纤维化 ,PPARγ蛋白和mRNA的表达减弱 ;中药治疗组肝组织脂变、炎症和纤维化程度显著减轻 ,接近正常对照组 ,PPARγ蛋白和mRNA的表达增强 (与治疗对照组比较P <0 0 5 ) ;治疗对照组血清ALT水平和肝细胞脂变程度较模型组减轻 ,但炎症、纤维化和PPARγ的表达无显著变化。结论 复肝解毒方可以有效逆转酒精所致的肝损伤 ,这与本方可以活化核因子PPARγ ,抑制酒精性肝损伤的炎症反应有关。

Objective To investigate at the molecular level the mechanisms of Fuganjiedu Formula (FF) in treating alcoholic liver disease.Methods 40 Wister rats were randomly divided into the normal control group (10 rats) and the model group (30 rats);all the rats in the normal control group and 10 rats in the model group (1 died of pulmonary choke during the experiment) were decollated at the end of the 16th week,and the samples of the serum and liver tissue were collected;the rest 20 rats in the model group were randomly divided into the TCM drug-treating group and treating-control group (10 rats in each group),and the rats in both groups were executed after 24 weeks.HE,Sudan IV and Masson's trichrome staining were respectively used to observe the microscopic pathologic changes and the changes in the electronical microscopic ultrastructures of the hepatic tissue,and the immunohistochemical staining and RT-PCR were used to observe the effects of FF on the expressions of peroxisome proliferator-activated receptor-γ(PPAR-γ) protein and gene.Results In the model group,the levels of ALT and TNFαin the serum and the level of MDA in the liver homogenate were increased;obvious adipose denaturalization,inflammation,necrosis and fibrosis were observed in the hepatic tissue;and the expressions of PPAR-γ protein and mRNA were decreased.In the TCM drug-treating group,the severity of adipose denaturalization,inflammation,necrosis and fibrosis of the hepatic tissue was markedly alleviated (which was close to that in the normal control group),and the expressions of PPAR-γ protein and mRNA were increased (P<0.05 as compared with those in the treating-control group).In the treating-control group,the level of ALT and the severity of hepatocellular adipose denaturalization were reduced as compared with those in the model group,but there was no marked change in the fibrosis and the expression of PPAR-γ.Conclusion FF can effectively reverse the alcoholic hepatic lesion,the mechanism of which is related with FF's activating PPAR-γ,and inhibiting the inflammatory reaction in alcoholic hepatic lesion.