| PrefaceCirrhosis is a final stage of various kinds of liver disases,which treatment is always explored for many years.Liver transplantation is a better method to cirrhosis,but owning to the problem of its expansive cost,limited donator and ethics,its extensive application is also limited.thus,to look for a simple new way to cirrhosis is a aim of study. Recently,as research of stem cell progress,application of marrow stem cells in clinic is gradually payed close attention.Basis studys founded marrow stem cells may differentiate into function liver cells in vitro,and it can grow and differentiate within surroundings of hepatic injury or fibrosis in vivo.meantime it can improve alnormal liver function.Many cirrhosis models were induced by injection carbon tetrachloride in rats,which has great difference to clinical application,but big animal cirrhosis model is immaturity.Now bone marrow stem cells have been applicated in animal experiment and clinic,such as myocardial ischemia,diabetes and liver diease et al.Study founded bone marrow stem cells transplantion has a better effect to hepatic injury and severe liver dieases,but it seems not satisfactory to cirrhosis.It is very necessary to search for a new method enhancing its effect.Our project is that autologous bone marrow stem cells were transplanted through the hepatic artery in rubbit's hepatic injury and cirrhosis model.and the safety and efficacy of autologous bone marrow stem cells transplantion were evaluated by detecting liver function and pathohistology,Besides,the affect of hepatocyte growth-promoting factor to stem cell transplantation to cirrhosis was observed. Part 1The Development of Carbon Tetrachloride-Induced Hepatic injury and Cirrhotic Model in RabbitsObjectiveTo search for a method of carbon tetrachloride-induced Hepatic injury and cirrhotic model in rabbit,and evaluate the feasibility and achievement ratio of the models through this method.Methods70 Male New Zealand white rabbits were divided into 3 group.The rabbits were subcutaneously injected with carbon tetrachloride once per day for 4 days in acute Hepatic injury model group,and liver pathology and function ware examined.Meanwhile,The rabbits were subcutane- ously injected twice weekly in cirrhotic model group,and liver pathology and function ware examined 12 weeks after the injection.ResultsIn acute Hepatic injury model group,after carbon tetrachloride administration,the enzyme activity was steped up.The number of ALT goes up from 35.83±8.04u/l to 331.00±29.15u/l,and TBIL goes from 6.41±0.52 umol/1 up to 11.64±0.58 umol/l。and the serious cellular necrosis was founded,and hepatic structure was alnormal.However,in cirrhotic model group,the albumn(ALB) and thrombozyme activity(PTA) decreased evidently compared to normal control group.The number of ALB decreased from 40.28±2.30g/l to 27.46±1.76g/l.and PTA decrease from 6.41±0.52 umol/l up to 11.64±0.58 umol/l。In pathohistology,we founded that the fiber hyperplasy was evident and typical pseudo-lobule can be seen in the rabbit liver,ConclusionA successive method to induce rabbit Hepatic injury and liver cirrhosis was developed.But the mortality is high in cirrhotic model group. Part 2Experimental Study of Autologous Bone Marrow Stem Cells for the Tr eatment of Acute Hepatic Injury in Rabbit.ObjectiveA rubbit model was used to evalute the feasibility of treating acute hepatic injury through transplanting autologous bone marrow stem cells via the hepatic artery.MethodsFifteen rubbits were induced to develop acute hepatic injury model by daily subcutaneous injection with 50%CCL4 0.8ml/kg for 4 days.Then the animals were random- ly divided into two experimental and control groups. In the experimental group,autologous bone marrow stem cells(1×10~8) were disassociated and transplanted into liver via the hepatic artery.Besides,in the second experimental group,the hepatocyte growth- promoting factor(pHGF) was given via vein injection with 2.0mg/kg every other day for 20 days.The change of liver functions had been monitored by biochemical methods in 2,4,8weeks and histopatho-logic examination at 8 weeks after transplantation.ResultsAfter CCL4 injection,hepatocyte necrosis was noted.An increase in AST,ALT and TBIL activity(P＜0.05) was also observed obviously.But the levels of TP,and ALB were not obviously changed(P＞0.05) in plasma. 8 weeks after stem cells transplantation,the activity of AST and ALT,and the levels of TBIL were lower than the control group(P＜0.05).and the levels of TP and ALB were higher than the control group(P＜0.05).Moreove,the above changes in the second experimental group were more obvious than that of the first experimental group(P＜0.05).In pathohistology,we founded that liver tissue structure in the experi-mental group was more close to normal than that of the control group,and fiber hyperplasy and hepatic cell degeneration was founded in the control group.Besides,the number of hepatic CD34+ cells of liver tissue in the experimental group was much than that of the control group.The number of the CD34+ cells can be seen per high power field of vision is 1～2 cells in the control group.3～5 cells in the first experimental group,and 6～8cells in the second experimental group.ConclusionIt is feasible that autologous bone marrow stem cells transplanted through the hepatic artery in the rubbit liver necrosis model,and the transplanted stem cells can regenerate in the acute injurey liver.The hepatocyte growth-promoting factor can help to improve hepatic function. Part 3Treatment of chronic hepatic cirrhosis with autologous bone marrow stem cells transplantationObjectiveTo evalute the feasibility of treatment for rubbit model with hepatic cirrhosis by transplantation of autologous bone marrow-derived stem cells via the hepatic artery.and the effect of hepatocyte growth factor for treatment of stem cells transplantation to liver cirrhosis.MethodsFourty-five rubbits were induced to develop acute hepatic injury model by daily subcutaneous injection with 50%CCL4 0.8ml/kg for 4 days.Then twenty-five animals were randomly divided into two exper- imental and control groups.In the experimental group,autologous bone marrow stem cells(1×10~8) were disassociated and transplanted into liver via the hepatic artery.Besides,in the second experimental group,the hepatocyte growth-promoting factor(pHGF) was given via vein injection with 2.0mg/kg every other day for 20 days.The change of liver functions had been monitored by biochemical methods in 4,8,12weeks and histopathologic examination at 12 weeks after transplantation..ResultsAfter transplantation of bone marrow stem cells,the liver function of patients improved.Eight weeks after transpl-anttation,the activity of AST and ALT,and the levels of TBIL were lower than the control group,and the levels of TPA and ALB were higher than the control group,but these changes have not obvious difference.However,the above changes in the second experimental group were more obvious than that of the first experimental group.And the difference between the second experimental group and control group was very significant(P＜0.05).In pathohistology,we founded that hepatic cells in the experimental group was more close to normal,but cell degeneration was founded in the control group,the fiber hyperplasy and hepatic pseudo-lobule was lessevident than that of the control group.Besides,the number of hepatic CD34+ cells of liver tissue in the experimental group was much than that of the control group.The number of the CD34+ cells can be seen per high power field of vision is 2～3 cells in the control group.3～6 cells in the first experimental group,and 6～9cells in the second experimental group.ConclusionIt is effective that autologous bone marrow stem cells transplanted via the hepatic artery for the rubbit liver injury model,and the transpl-anted stem cells can regenerate in the acute injurey liver.The hepatocyte growth-promoting factor can help to improve hepatic function. |
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