茵陈蒿汤干预二甲基亚硝胺大鼠肝硬化效应的方证病理学基础研究

Recipe-syndrome Correlation and Pathogenesis Mechanism of Yinchenhao Decoction in Intervening Dimethylnitrosamine Induced Liver Cirrhosis Progress in Rats

目的基于清热利湿与滋养肝肾不同功效古典方剂的干预效应,探讨二甲基亚硝胺(dimethylni-trosamine,DMN)诱导大鼠肝硬化的方证病理学基础。方法腹腔注射DMN4周制备大鼠肝硬化模型,造模2周末取6只模型大鼠作给药前观察,其余模型大鼠随机分为模型对照组、茵陈蒿汤(yinchenhao Decoc-tion,YCHD)组及一贯煎(yiguanjian,YGJ)组;各组于继续造模同时给予不同方剂的煎剂经口灌胃2周,正常大鼠与模型对照组给予同量等渗盐水,4周末处死大鼠,获取血清与肝组织样品,观测肝功能、肝组织病理学、α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)与羟脯氨酸(hydroxyproline,Hyp)含量,检测肝组织丙二醛(malondialdehyde,MDA)、谷胱甘肽(glutathione,GSH)含量,谷胱甘肽S转移酶(glutathione S-transferase,GST)活性;免疫印迹检测肝组织α-SMA、肿瘤坏死因子α(tumor necrosis factor-alpha,TNF-α)、血小板衍生生长因子(platelete-derived growth factor,PDGF)以及肝脂肪酸结合蛋白(liver fatty acid bindingprotein,L-FABP)、转铁蛋白(transferrin,Tf)表达的变化。结果与模型对照组比较,YCHD组肝功能显著改善;肝组织Hyp含量与α-SMA蛋白表达显著降低(P<0.05),显著抑制DMN大鼠肝硬化的形成,而YGJ无明显作用。肝组织TNF-α与PDGF表达在YCHD组显著减少,而YGJ组TNF-α进一步增加,PDGF无明显变化;YCHD组肝组织MDA含量与GST活性显著降低(P<0.01或P<0.05),GSH含量、L-FABP及Tf蛋白表达显著增加(P<0.05),而YGJ组仅见GSH显著增加(P<0.05)。结论茵陈蒿汤显著抑制DMN大鼠肝硬化的形成;DMN大鼠肝硬化形成期的肝组织炎性病变及其过氧化损伤是茵陈蒿汤清热利湿效应的方证病理学基础。

Objective Lay on the intervention effect of different classic Chinese medicine recipes for clearing-heat,eliminating dampness and nourishing Gan-Shen,to investigate the recipe-syndrome pathologic foundation of dimethylnitrosamine（DMN） induced liver cirrhosis in rat.Methods Liver cirrhosis model rats were made by DMN intra-peritoneally injection for 4 weeks at the dosage of 10 mg/kg body weight,once per day for 3 consecutive days in each week.Excepting the 6 rats for pre-treatment observation,others were divided into 3 groups and gastric infused respectively with saline（model group）,Yinchenhao Decoction（YCHD group） and Yiguanjian（YGJ group） in the continuous 2-week modeling period.Besides,a normal group was set up with 10 healthy rats administered by saline.At the end of the 4th week,rats were sacrificed and their blood sample and liver tissue were get for detecting liver function;hepatic histology（HE and Sirus red stain）;α-smooth muscle ac-tin（α-SMA） and hydroxyproline（Hyp） contents,Oxidative stress-related parameters such as malondialdehyde（MDA）,glutathione（GSH）,and glutathione S-transferase（GST）;expressions of α-SMA tumor necrosis factor-alpha（TNF-α）,platelete-derived growth factor（PDGF）,liver fatty acid binding protein（L-FABP）,and transferrin were determined at the same time.Results Compared to those in the model group,liver function was improved significantly and content of Hyp,α-SMA protein expression decreased remarkably in the YCHD group（P 0.05）,showing significant inhibition on liver cirrhosis formation;while those effect was not shown in the YGJ group.Comparisons of various parameters between groups showed that TNF-α,PDGF expression,MDA content and GST activity in liver tissue were decreased and GSH content,L-FABP and transferrin expression were increased significantly at the terminal of the experiment in the YCHD group（P 0.05）,but only increases of TNF-α and GSH were shown in the YGJ group（P 0.05） .Conclusion YCHD exerts significant inhibition on DMN-induced cirrhosis formation in rats,and the inflammatory change and peroxidation in liver tissue at liver cirrhosis developing stage of DMN treated rats is the recipe-syndrome pathologic foundation of clearing heat and eliminating-dampness effects of YCHD.