Objective:Dahuang Zhechong recipe（DHZC） is a traditional Chinese formula for treatment of cirrhosis.The present study was conducted to investigate the effect of ultramicro-powder of DHZC on rats liver fibrosis and its possible mechanisms by Two-dimensional differential in-gel electrophoresis（2D DIGE）based proteomics analysis.Method: SD rats were randomly divided into 3 groups: normal group

model group and DHZC-treated group.Except in normol group

liver fibrosis in rat was produced in each group.The rats were injected intraperitoneally with 0.5 mL sterile porcine serum twice a week for 12 weeks.In addition

rats of normal group treated with physiological saline in the same way.At the same day

aqueous solution of ultramicro-powder of DHZC was given to the DHZC-treated group（0.27 g · kg-1

ig） for 12 weeks.Rats of three groups were given equal volume of physiological saline.The rats were killed after 12 weeks of porcine serum treatment.Blood samples of all groups were drawn from the abdominal aorta.Livers were stained with HE and Masson to observe the histopathological chang.The lever of serum alanine aminotransferase（ALT）

aspartate aminotransferase（AST）

hyaluronic acid（HA）

laminin（LN）

procollagen type III（PC-III） and collagen type IV（IV-C） were measured.The difference gel electrophorisis（2-D DIGE） combined MALDI-TOF MS/MS technique was used to analyze differentially expressed proteins of rat livers from model group and DHZC-treated group.Result: Rats of model group showed significantly increased serum ALT

AST

HA

LN

PCⅢ and IV-C levels compared with rats of normal group（P<0.01）.The histopathological analysis suggested that the severe liver fibrosis happened in model group compared with normal group.Administration with ultramicro-powder of DHZC obviously alleviated the degree of liver fibrosis based on HE stained and Masson stain tissue sections.Fifty-eight spots representing significant changes in protein expression were detected utilizing DIGE.Twelve of these proteins were send to identify by MALDI-TOF MS/MS.Two proteins were confirmed by Western blot and immunohistochemistry.The contents of ALT

AST

HA

LN

PCⅢ and IV-C in model rats were increased significantly

while DHZC could decrease those significantly.The histopathological analysis suggested that DHZC obviously alleviated the degree of liver brosis induced by porcine serum.Conclusion:DHZC can effectively prevent hepatic fibrosis and improve liver function in rats.The regulation on the expressions of some proteins may be one of the pathways for DHZC to exert its anti-hepatic fibrosis effect.