Influence Of Hao Bie Yang Yin Ruan Jian Fang On Hepatic Extracellular Matrixes In Rats With Alcoholic Liver Disease

Objective:To study the effect of Hao Bie Yang Yin Ruan Jian Fang(HBYYRJF)on alcoholic liver disease in model rats,and confirm preventive effect of HBYYRJF on experimental hepatic fibrosis and investigate the possible mechanisms by degrading the synthesis of extracellular matrixes of liver,to provide experimental basis of hepatic fibrosis to clinical use of HBYYRJF.Methods:100 healthy Wistar male rats,weighted 200-240g,were divided into 5 groups by random:high dose and low dose HBYYRJF groups,20 rats in each group;positive control group(colchicine:20 rats),model group(32 rats);control group(8 rats).The model of alcoholic liver disease was established by intragastric administration alcohol and feeding fat diet,pyrazole and carbony iron as well.Alcohol was administered in HBYYRJF and colchicine groups respectively.Dosage:high dose of HBYYRJF:8.2g·kg-1·d-1;low dose of HBYYRJF:2.59 g·kg-1·d-1;colchicine group:0.09mg·kg-1·d-1;alcohol:6.2g·kg-1·d-1(contain 1.2ml·100-1·g-1·d-1);distilled water:1.2ml·100-1·g-1·d-1.The preventive groups were treated by prophylactic medication.Drugs were given to the corresponding therapeutic groups.The therapeutic groups:alcohol was given in the morning and drugs were given in the afternoon;The model group:alcohol was given in the morning and distilled water was given in the afternoon;The normal group:distilled water was given in the morning and in the afternoon.After ten weeks,rats were sacrificed,and samples were taken.To count the liver weight,calculate the ratio of body weight and liver weight in each group.Pathological changes of Hepatic tissue and the degree of hepatic hyperplasia were observed under light microscope.Liver tissues were used to measure the content of hydroxyproline by acid hydrolysis,and then converted to collagen protein.Serums were used to measure the contents of MDA,HA,LN,IV-C?PC? by radio-immune method.The level of ALT and AST in Serum were measured by spectrophotometer.The expression of TIMP-1 were measured by western blot and the level of MMP-13 in liver tissue were measured by immunocytochemical method.Results:1.Statistics of rats in each group between the liver weight,body weight,liver weight body weight ratio and death rate:The body weight in model group was significantly lower compared with normal group(P<0.01),there was no significantly difference in liver weight between all groups.The ratio of liver weight and body weight in model group was significantly higher compared with normal group(P<0.05),and there was no significantly difference between therapy groups and normal group.The death rate of rats in model group was significantly increased compared with normal group(P<0.01).The death rate in therapeutic groups was significantly decreased compared with model group.2.Histological examination and pathological grading:HE staining:fatty liver and hepatitis were found in model group.An obvious improvement was found in hepatic tissue in HBYYRJF prescription groups.The degree of hepatic fibrosis in model group was significantly severe compared with normal groups(P<0.01),the therapeutic groups were significantly decreased compared with model group rats,there was no significantly difference between therapy groups and model groups.3.The level of collagen protein:The collagen protein in model group was significantly increased compared with normal group rats(P<0.01);the collagen protein in therapeutic groups were significantly decreased compared with model group rats(P<0.05).4.The contents of IV-C?PC??HA and LN:there was no significantly difference in all groups(P>0.05).5.The level of TIMP-1?MMP-13:TIMP-1?MMP-13 in model group was significantly increased compared with normal group rats;TIMP-1?MMP-13 in therapeutic groups were significantly decreased compared with model group rats.Conclusion:1.The model of hepatic fibrosis of rat was successful2.Haobieyangyinruanjianfang has a prophylaxis effect on alcoholic liver disease3.The mechanism of its effects might be associated with decreasing the expression of TIMP-1 and MMP-13,promoting degradation of extracellular matrixes.