Establishment Of The Rabbit Hepatic Fibrosis Animal Model And Treatment Of Hepatic Fibrosis By Transplantation Of Autologous Bone Marrow Stem Cells

Objective:This research is to establish the hepatic fibrosis model induced by Carbon Tetrachloride(CCL₄)in rabbit.Autologous bone marrow mononuclear cells (BMMNCs)which isolated and labeled in vitro were transplanted into hepatic fibrosis animal through cranial mesenteric vein(CMV).We observe Whether the transplanted cells can home to the injured hepatic tissue and accelerate the regeneration and reparation of the injured liver and improve hepatic function.Methods:1 Establishment of the hepatic fibrosis animal model:the rabbits were injected with 40%CCl₄-olive oil(1～1.5ml/kg I.H.2 times per week,last 12 weeks),until pathology showed hepatic fibrosis.2 Separation of BMMNCs by density gradient centrifugation and labeling by BrdU.3 Transplantation of labeled autologous BMMNCs into hepatic fibrosis animal through cranial mesenteric vein.4 Judgment of the curative effect after cellular transplantation:The measurement of hepatic function parameters,pathological investigation and immunohistochemistry analysis of hepatic tissue were carried out on the 12th week of making model and on the 3th,7th,14th,21th day after cellular transplantation.Result:1 Histopathological changes of animal model:Histopathological observation show that lobules of liver in the rabbits of model group was disorder with inflammatory cell infiltration and fatty degeneration of hepatocytes.While collagens were deposited around the centralveins and portal areas and the fibrous septa were formed on the 12th week of injection with 40%CCl₄-olive oil.2 Change of liver function:On the 21th days of autologous BMMNCs transplantion,compared with control group,activity of ALT and AST in serum decreased significantly(P＜0.05),the content of TBIL and DBIL in serum decreased significantly(P＜0.05)while the content of ALB in serum increased significantly(P＞0.05)in the transplantation group.3 Liver histopathological changes after cellular transplantation:After transplantion of autologous BMMNCs,With the time passing by,pathology showed that the fibrosing hepatic tissue was repaired gradually,which was repaired better in the transplantation group than in the control group in the same time.4 Distribution of transplanted cells in liver:Transplanted cells showed that BrdU staining were positive around the centralveins of liver lobules in the third day of cell transplantation and last to the end of the experiment,While the control group BrdU staining was negative.Conclusion:1 The typical rabbit model of the hepatic fibrosis was produced by subcutaneous injection of 40%CCl₄-olive oil for 12 weeks.2 BMMNCs which transplanted through CMV can home to the fibrotic tissue of liver.3 BMMNCs transplantation can decrease activity of ALT and AST,decrease content of TBIL and DBIL,increase content of ALB in serum,and improve liver function.