Rosmarinci Acid Improved Liver Fibrosis In Mice By Regulating Macrophage Polarization

Objective:Hepatic fibrosis occurs as the consequence of diverse hepatic diseases,such as virus hepatitis,alcoholic liver disease,nonalcoholic fatty liver disease,autoimmune hepatitis,and it is characterized by excessive deposition of collagen fibril leading to ultimate irreversible cirrhosis.About 500 thousands deaths a year from hepatic cirrhosis and its complications in China.But there is a few of effective treatments against hepatic fibrosis.Mass research results demonstrated that rosmarinic acid has the functions of anti-inflammation,anti-oxidantion and anti-ischemia.And a study indicated that rosmarinic acid can alleviate hepatic fibrosis.Furthermore a French scholar showed that macrophage is associated with hepatic fibrosis.The aim of this study is to confirm the beneficial effect of rosmarinic acid to hepatic fibrosis,and study the mechanisms related to macrophage.Methods:1.In vivo experimentThe model of hepatic fibrosis was established by used in the intraperitoneal injection with CCl₄,10ml/kg two times a week,and then determinated the contents of serum ALT and AST.The expressions of?-SMA,Col????,Col????in liver were measured by Real-time PCR,which represent levels of liver fibrosis.In additional our study measured expressions of M1 and M2 markers with Real-time PCR.2.In vitro experimentMurine macrophages cell lines J774A.1 was cultured in DMEM.Cells were treated with 1ng/ml of LPS and 5ng/ml of IL-4,and then M1 and M2 were treated with rosmarinic acid 27um.At the same time the levels of M1 and M2 markers in each group were determined by Real-time PCR.Results:1.In vivo experiment1.1.The levels of ALT and AST in serum of model group were higer than control group,and the levels decreased by treated with rosmarinic acid.The compare between experimental group with model group has statistical significance?P?0.05?.1.2.The expressions of?-SMA,Col?1?I?and Col?1???in liver of model group were higer than control group,and the expressions decreased by treated with rosmarinic acid.The compare between experimental group with model group has statistical significance?P?0.05?1.3.The expressions of M1 makers?TNF-?,IL-1?,NOS2?and M2 makers?Arg-1,Mrc2?increased in model group.After treated with rosmarinic acid,the expressions of M1 and M2makers decreased,and the compare between experimental group with model group has statistical significance?P?0.05?.2.In vitro experimentAfter treated with IPS and IL-4 in J744A.1 cells respectively,the levels of M1 and M2maker raised evidently.And the expressions of M1 makers?TNF-?,IL-1?,NOS2?decreased after treated with rosmarinic acid.The compare between experimental group with model group has statistical significance?P?0.05?.However the expressions of M2 makers have not change much,except for the level of Arg-1.Conclusion:Rosmarinic acid can ameliorate CCl₄ induced liver fibrosis in mice,and the effect probably is associated with polarization of macrophage.