The Preliminary Study Of Gexiazhuyutang Regulate Endothelial Progenitor Cells To Build Liver Fibrosis Micro-environment

Background and aims:Liver fibrosis is the result of extracellular matrix over production and abnormal accumulation after chronic liver injury. Nearly all chronic liver disease include chronic hepatitis B(C), chronic schistosomiasis, Chronic alcohol and drug damage, autoimmune liver disease always follow the pathological change. So the research of liver fibrosis plays a important part in treatment of liver disease. Endothelia progenitor cells is the precursor of endothelia cells. They belong to multipotent stem cells. They always transfer to injured part and proliferate and differentiate into mature EPs with the induction signal’s guidance, and then take part in the rebirth and repair of blood vessel. In 1997, Asahara first used immunomagnetic beads to separate EPCs from human peripheral blood. After that EPCs plays a important part in different areas like peripheralvascular disease, cardiovascular and cerebrovascular diseases, the formation of tumor capillaries, healing of wound. And it rose a new way to treat different kinds of diseases. And with the clinical observation, I found the mechanism of Gexiazhuyutang had a good efficacy of the treatment for liver fibrosis. So this project makes a connection of Chinese medicine and western method, and pay attention to the experiment that with the intervention of the mechanism of Gexiazhuyutang, how could EPCs rebuild liver fibrosis micro-environment.This project connects Chinese clinic treatment with western experimental research methods, and aims at finding a new method to treat liver fibrosis.Method:(1)ICR rats intraperitoneal injection carbon tetrachloride to build liver fibrosis model for 6 weeks. Then give the succeed model gavage of the mechanism of Gexiazhuyutang for 10 weeks. Text the blood and liver regularly and find the result of the treatment with the mechanism of Gexiazhuyutang. (2)C57 rats intraperitoneal injection carbon tetrachloride to build liver fibrosis model for 6 weeks. Then give the succeed model gavage of the mechanism of Gexiazhuyutang for 8 weeks.During the gavage, text the peripheral blood regularly with the method of flow cytometry to assess the EPCs of transfer from marrow to peripheral blood. (3)C57 rats intraperitoneal injection carbon tetrachloride to build liver fibrosis model for 6 weeks. Then give the succeed model gavage of the mechanism of Gexiazhuyutang for 8 weeks.During the gavage, separate the EPCs from peripheral blood and text them with colony forming experiment to assess the number of EPCs in peripheral blood.Result:(1)Liver function:After 6-weeks of intraperitoneal injection carbon tetrachloride, the result of AST, ALT and TBIL had tend to the top level. With the gavage of the mechanism of Gexiazhuyutang, each targets get lower. (2) HE staining and Masson staining:In the first weeks of molding, collagenous fiber slowly gathered to portal areas, and slowly changed into fibrous septum, and even turn to pseudolobuli disorder. But after the gavage, the collagenous fiber obviously reduced and the liver turned to better.(3) Flow cytometry:In the process of the gavage, the percentage of EPCs in all cells rose with the time going. (4)Colony forming experiment:In the process of the gavage, the transformation of EPCs from marrow to peripheral blood became more energetic.Conclusion:(1)The mechanism of Gexiazhuyutang has an obvious treatment to liver fibrosis. (2) The mechanism of Gexiazhuyutang can stimulate EPCs to treat liver fibrosis.