| objective: This experiment aims to investigate the effect of Huazhuo Jiedu Decoction on ethanol-induced liver in rat NF-κB, TNF-αlevel and Liver function,CAT, Hepatic fibrosis in the method of oral administration of ethanol gradient lavage to set up alcoholic liver disease (ALD) rat models which were lavaged with zhuo Jiedu Decoction , with the Combination of histopathological examination of liver detoxification method Huazhuo study in the treatment of alcoholic liver injury to explore the NF-κB, TNF-αin the pathogenesis of ALD in the role for the application of Huazhuo Jiedu controlling the disease and provide a theoretical basis.Methods: healthy 50 male Wistar rats, weighing 180±20g, normal diet a week after acclimation then randomly selected 12 cases as the normal group (normal diet and given saline 2.0ml / only, administered three times a day, a total of 12w). The remaining 48 cases integrated manufacturing rat model of alcoholic liver disease. On daily morning the rats were gavaged with alcohol dairy (Red Star brand Erguotou 56°, Beijing factory), the liquor initially be diluted 1%, 3%, 5%, 8%, four kinds of concentrations from low to high, daily gavage 2ml, the 3d. The first 13 days beginning, gavage according to liquor dose 5g/(kg.times) subsequently according to rat body weight, activity and response increase dose 10d one times, finally gavage dose 8g/(kg. times) for eight weeks, by gavaging corn oil at the same time 1ml/one/day. After the success of the model the rats were randomly divided into four groups: model group (MD), Ganlixin Group(diammonium glycyrrihizinate, DG), Huazhuo Jiedu Decoction high dose group (HZG), Huazhuo Jiedu Decoction of low-dose group (HZD). Each group was fed ordinary particles, free water and feeding, sub-cage reared at 22±2℃, humidity 70%±5% clean class animal laboratory . 24 over the weekend with femoral vein phlebotomy use, collecting serum for determination of the following items: alanine aminotransferase ALT, aspartate aminotransfera-se AST, Radioimmunoassay detection of tumor necrosis factor-α(TNF-α), laminin (LN), IV collagen (IV-C),Hyaluronic acid (HA), Procollagen typeⅢ(PCⅢ),Catalase(CAT); another 10% formalin-fixed, conventional embedding, slicing, conducted Hematoxylin --eosin (HE) staining to observe the basic pathological changes of liver tissue. By immunohistochemical staining of liver NF-κB expression level. Statistical treatment, all data are used (x_±s)express, using test analysis of variance, the application data achieved by statistical software SPSS 11.5.Results: 1 generally: model 8 rats killed and normal control group survived. compared to the initial weight of each group there were no significant differences to the srvival of rats.General performance: normal control rats, loss of appetite good, hair smooth, good mental state, weight gain. Model rats make them apathetic, reduced activity, anorexia, weight loss, dull coat, weight over the previous decline. Treatment of rats in each group spirit, appetite and activity between the normal group and model groups.2 Determination of serum biochemical indicators results: Compared with the normal group, model group, serum AST, ALT, PCⅢ, IV-C, HA, LN, TNF-αconcentration increased significantly (P <0.01), CAT was significantly decreased ( P <0.01); compared with the model group, the treatment group, serum AST, ALT, PCⅢ, IV-C, HA, LN, TNF-αdecreased were significantly different (P <0.01), CAT was significantly or high (P <0.01); Western medicine group, Huazhuo Jiedu Decoction between the low dose groups had no significant difference (P> 0.05); Huazhuo Jiedu Decoction high-dose group and WM group were significantly different (P <0.01).3 .Pathological changes in the liver: observation with the naked eye: normal rat liver capsule smooth, soft texture, Red-brown. Model rats significantly increased liver size and texture than the normal hard, bleak than color, light-yellow or yellow soil, yellow and white can be seen with focal lesions degeneration, greasy section. with light microscope the treatment group that between normal group and model groups, liver biopsy showed that rat model liver cells with varying degrees vacuolar degeneration, focal necrosis of liver cells sinus volume can be seen more multinucleated cells, lymphocytes and mononuclear cell infiltration, hepatocellular swelling was balloon-like change, cells filled with lipid droplets of various sizes, within the loose cytoplasm, numbers of fat vacuoles of varying nucleus were pushed to one side, the most obvious fatty degeneration of liver cells is in the central area of liver cells . compared to model control group,Chinese medicine and western medicine treatment group have varying degrees of improvement and recovery.4. The immunohistochemical staining results of NF-κB in liver tissue:NF-κB positive expression in the liver was mainly located around the central vein of the cytoplasm and cell nucleus. Normal NF-κB did not express basically; model rat liver tissue expression of NF-κB-positive liver cells significantly more than the control group, there was not only the expression of the cytoplasm, ,but also a clear change of granular brown in the majority of nulei; Treatment of each group the expression of NF-κB reduced, to one of medicine the most high-dose group decreased significantly, 24 the weekend of the expression of NF-κB near normal.Conclusion:1 the way of adopting a gradual increase in ethanol concentration of rats with gavage method is successful for the copy rat model of ALD, the pathological changes reflect a clinical alcoholic liver disease (such as: fat accumulation, inflammation, fibrosis). 2 Immunohistochemistry confirms that NF-κB plays an important role in the pathophysiology process of ALD, in the experiment it involved in the ALD process of liver injury in rats and involved in regulation of cytokines and inflammatory factors such as gene expression.3 Huazhuo Jiedu prescription can inhibit NF-κB, TNF-αexpression in alcoholic liver disease, and improve liver function and liver fibrosis target, therefore, involving Huazhuo Jiedu in the treatment of ALD has practicability and feasibility.
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