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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 explore the mechanism of traditional Chinese compound Fugan Jiedu Decoction in the treatment of alcoholic liver disease at the molecular level. Methods A total of 40 Wistar rats were randomly divided into normal control group (10 rats) and model group (30 rats). The normal control group (10 rats) and the model group rats (9 rats) were sacrificed at the end of the 16th week. Serum and liver tissue specimens. The other 20 rats in the model group were randomly divided into a Chinese medicine treatment group and a treatment control group, 10 rats in each group, and were sacrificed after 24 weeks. HE, Sudan IV and Masson trichrome staining were used to observe the pathological changes of liver tissue under light microscope and the ultrastructure under electron microscope. The effect of Fugan Jiedu Fang on the expression of peroxisome proliferator activated receptor (PPARγ) protein and gene was observed by immunohistochemical staining and RT PCR. Results The levels of serum alanine transferase (ALT), tumor necrosis factor (TNFα) and malondialdehyde (MDA) in liver homogenate were increased in the model group, and the hepatic tissues showed marked steatosis, inflammation, necrosis and fibrosis. The expression of PPARγ protein and mRNA was attenuated. The degree of liver tissue lipidosis, inflammation, and fibrosis were significantly reduced in the Chinese herb treatment group. Compared with the normal control group, the expression of PPARγ protein and mRNA was increased (P < 0.05 compared with the treatment control group). The serum ALT levels and hepatocyte lipidosis in the treatment control group were less than those in the model group, but there was no significant change in the expression of inflammation, fibrosis, and PPARγ. Conclusion Fugan Jiedu Decoction can effectively reverse the alcohol-induced liver injury, which is related to the activation of nuclear factor PPARγ and inhibition of the inflammatory response of alcoholic liver injury.