Allogeneic Hepatocytes Transplantation Intraperitoneally And Augmentor Of Liver Regeneration To Treat Acute Liver Failure In Rats

Objective: Acute liver failure (ALF) is onset emergency.Its natural mortality is high. The most effective therapy for ALF is orthotopic liver transplantation. The shortage of livers has given rise to transplant waiting lists that are several years long, yet it has also stimulated intensive research to identify novel therapies for liver disease. Hepatocytes transplantation (HCT) is generally accepted as an effective, safe and promising therapy. Intrasplenic transplantation is the most effective pathway manifested by animal experiments. But it is dangerous to inject hepatocytes into spleen in clinical operation.In this study, our objective is to investigate the effect of allogeneic HCT with augmentor of liver regeneration (ALR) intraperiton- eally on ALF induced by D-gal, to determine the role of ALR in alloimmunity.Methods: Hepatocytes of adult SD rats were isolated with semi-in-situ collagenase perfusion method. 74 ALF rats were induced by D-ga(l1.0g/kg). HCT was received intraperitoneally 24h after intoxication,and the rats were divided into five groups randomly:GroupⅤ14 rats and other groups 15 respectively.GroupI received normal saline (NS). GroupⅡreceived 2×107 hepatocytes and NS. GroupⅢreceived 2×107 hepatocytes and CsA(10mg.kg-1.d-1). GroupⅣreceived 2×107 hepatocytes and ALR (50ug.kg-1.d-1). GroupⅤreceived ALR(50ug.kg-1.d-1). The survival rate, pathohistological examination of transplanted hepatocytes and liver, the levels of ALT,AST,TBil,CD4,CD8,CD68,IL-1βand TNF-αof all rats were observed.Results: Comparison two-week survival rate with each other group, GroupⅣwas higher than that of GroupⅠ(66.7% vs 0%,P=0.001). Survival rates of other groups were similar. Serum biochemical indicators ascended to the highest levels on second or third day after drug administered.And there were no differences between the indexes of all groups.But ascitic d1-2 ALT of groupⅢwas higher than that of groupⅠ,ⅣandⅤ.Ascitic d1-2 AST of groupⅡandⅢwas higher than that of groupⅣandⅤrespectively, and groupⅠwas higher than groupⅣ. Ascitic d1-2 TBil of groupⅠwas lower than that of groupⅡandⅣ. Biochemical indicators returned to the normal levels on 14th day post transplantation. Serum d1-2 TNF-αof group I andⅡwas higher than that of groupⅣandⅤ, and groupⅢwas higher than groupⅤ. Serum d1-2 IL-1βof groupⅣwas lower than that of groupⅡ.The levels of ascitic d1-2 TNF-αand IL-1βof groupⅡwere higher than that of other groups. Positive rate of d1-2 CD68 of groupⅡwas higher than that of group I andⅣin greater omentum. And positive rates of d1-2 CD68 of groupⅡandⅢin hepatocytes nodus were higher than that of groupⅣ. Positive rates of d1-2 CD4 and CD8 of groupI andⅡwere higher than that of groupⅣandⅤ. Positive rate of groupⅡd1-2 CD4 was higher than that of groupⅢ. All inspection items of survival groups were similar on 14th day after transplantation.Conclusion: After HCT and ALR intraperitoneally,the survival rate of ALF rats intoxicated with D-gal was increased. ALR can suppress alloimmunity effectively. ALR improved liver function and histology, and promoted survivorship and proliferation of transplanted hepatocytes.