Experimental Study On Alcoholic Liver Fibrosis By FFBJRGP

Objective In China, alcoholic liver fibrosis patients have been on the rise recently. It has been reported that about 20 percent of people, who have 10 to 20 years of alcohol drinking, might exhibit alcoholic liver fibrosis. Nowadays, alcoholic liver fibrosis, as an independent disease type, is focused extensively by researchers. At present, researches on pathological mechanisms of alcoholic liver fibrosis were seldom, many mechanisms were undefined yet. So based on the alcoholic liver fibrosis rat model, we investigated the major pathological mechanisms of experimental alcoholic liver fibrosis and discuss the pathogenesis and biological basis of hepatic collateral disease (LUO BING) caused by toxin invasion hepatic collatera (GAN LUO) in order to supply the basic theory and experimental data for prevention and treatment of alcoholic liver disease by Traditional Chinese and Western Medicine combination. Methods 1. The alcoholic liver fibrosis rat model was induced by intragastric infusion of a mixture of alcoholic liquid which included alcohol, conventional corn oil and pyrazole (1000:250:3), combined with a high fatty food as previously described over a period of 16 week. We examined the concentration of ALB, TP, AST, ALT, HA and LN in serum respectively in 4, 8, 12, 14 and 16 week later. Animals were sacrificed, and livers were harvested for morphology inspection. The part samples were formalin-fixed, embedded in paraffin and stained with hematoxylin and eosin, And Mallory stain, sirius red stain were used for evaluate fibronectin and collagen expression. The other part samples were snap-frozen in melting isopentane and stained with oil red O and hepatin PAS-Shiff. 2.The level of GSH, MAD, and HYP in liver tissue was analysed by some methods as previously described in early stage of alcoholic liver fibrosis. To observe the ultrastructure of hepatic sinusoid and sinusoid endothelial cell (SEC) with the ultra-lamina slice and electron microscopy. The typeⅠ , Ⅳcollagen and LN protein in liver tissue were visualized by immunohistochemistry stain. MMP-9mRNA and TIMP-1mRNA were measured by in situ hybridization and TGF-β1 protein content in liver tissue was analyzed by western-blot. Furthermor, we freshly isolated and purified the Kupffer cell and tested the expression of TNF-α and TGF-β1 protein. 3.After challenging, the mice were divided into 6 groups, those are control group, untreated model group, Colchicine Houde treated group, high dose of FFBJRGP treated group, media dose of FFBJRGP treated group, low dose of FFBJRGP treated group. After the 8w treatment, animals were sacrificed and the contents of ALB,TP,AST,ALT,ColⅣ,LN in serum were analysed, as well as the GSH,MAD, HYP in liver tissue. The pressure and the diameter of portal vein were also examined. The part samples were formalin-fixed, embedded in paraffin and stained with hematoxylin and eosin. Mallory stain were used for evaluate collagen. The type Ⅳ collagen and LN protein in liver tissue were visualized by immunohistochemistry stain. The other part samples were homogenated to analyse the protein of TGF-β1. 4. Spss 10.0 was used. Data were analyzed using one-way ANOVA. All values were expressed as mean ± SE. Statistical significance was assigned at a P value of <0.05 or <0.01. Results 1. The proportion of AST/ALT and HA content in serum raised in the startup or progress stages of ALD. 2. The lipid, carbohydrate and protein in hepatic cell decompensate in the progress of ALD. 3. The mechanism of liver fibrosis induced by alcohol are different form CCl4, the major pathological changes of alcoholic liver fibrosis are hepatic sinusoid capillarization and perisinusoid fibrosis. 4. The activation of Kupffer Cell is the critical molecule event of the startup of alcoholic liver fibrosis and high expression of TNF-α is a better marker of activation of Kupffer Cell. In the early stage of alcoholic liver fibrosis, Kupffer Cell is the main source of TGF-β1. 5. The free radical injury, the overexpression of basement membrane and the un-graduat...