| Part 1The Model Construction of Rats Underwent Partial Hepatectomy Liver AutotransplantationObjective:To probe the techniques for model construction of rat's partial hepatectomy residual liver autotransplantation and to provide a reliable model for clinical and basic researches on liver autotransplantation.Methods:150 cases of male Sprague-Dawley(SD)rats,among which 116 cases underwent formal operation,were taken to construct the partial hepatectomy residual liver autotransplantation model.Traditional venous-venous bypass(VVBP)was abolished and cold-perfused way was modified in our new techniques.Results:In 120 cases formal operations,The anhepatic time was 16～20(Med time 18.25±2.12)min and successful rate was 96.7% (116/120).time taken to vein tube implanted was 1～2min and 2/3 liver of rats' was resected in the model. Conclusion:The model modified by available technique,which is characterized with shorter operation time and better reproducibility,is particularly suit for researches referred to the effect of exogenous factors on residual liver after liver autotransplantation and is a reliable model for clinical and basic research of partial hepatectomy residual liver autotransplantation.Part 2The effects of Curcumin preconditioning on ischemia reperfusion injury after liver autotransplantation.Objective:To investigate the protective effects on the ischemia reperfusion injury(IRI)and the impacts on the neutrophil infiltration, expression of TNF-αand ICAM-1 by Curcumin preconditioning in rats after partial hepatectomy and residual liver autotransplantation.Methods:Male SD rats were used and available techniques were modified and simplified to construct the model.The rats were divided into three groups(Curcumin preconditioning(CU)group and 0.5% sodium carboxymethycellulose preconditioning(CM)group)randomly according to the different drugs administrated before residual liver reperfusion and the sham operation(SO)group was taken for control.6 cases animals of each group were killed at 2h,6h and 24h after residual liver reperfusion respectively.Plasma samples were collected for ALT and AST test;Liver tissues were harvested to detect histological change, MPO activity,the expression of the protein and mRNA of TNF-αand ICAM-1 by Immunohistochemistry and RT-PCR respectively.Results:The serum level of ALT and AST in CU group were lower than CM group significantly 2h,6h and 24h after reperfusion;The MPO activity of CM group was higher than CU group significantly at 6h after reperfusion(P＜0.05);Compared with CM group,the expression of TNF-αand ICAM-1 mRNA was decreased significantly in CU group 2h after reperfusion(P＜0.05)and the expression of TNF-αand ICAM-1 protein was reduced significantly in CU group 6h after reperfusion(P＜0.01,P＜0.05).Conclusion:High expression of TNF-αand ICAM-1,neutrophil infiltration may play an important role in the IRI process after rats' liver autotransplantation.Curcumin preconditioning can attenuate liver IRI and the mechanism may be associated with decreasing the expression of TNF-αand ICAM-1,inhabiting neutrophil infiltration. Part 3The effects of Curcumin preconditioning on liver regeneration after liver autotransplantation.Objective:To investigate the protective effects on the liver regeneration and the impacts on the apoptosis,expression of Bcl-2,TNF-αand NF-_κB by Curcumin preconditioning after partial hepatectomy and residual liver autotransplantation.Methods:Male SD rats were used and available techniques were modified and simplified to construct the model.The rats were divided into three groups(Curcumin preconditioning(CU)group and 0.5% sodium carboxymethycellulose preconditioning(CM)group)randomly according to the different drugs administrated before residual liver reperfusion and the hepatectomy(HE)group was taken for control.4 cases animals of each group were killed at 6h,24h,48h,72h and 168h after residual liver reperfusion respectively.The residual liver of each group animals were harvested and the quality were collected to count the regenerative rates;Parts of the liver tissues were harvested to detect histological change,apoptosis(TUNEL),mitosis,the expression of the protein and mRNA of Bcl-2,TNF-αand NF-_κB by Immunohistochemistry and RT-PCR respectively.Results:The regenerative rates of CU group were higher than CM group significantly from 24h to 168h after reperfusion(P＜0.01);The mitotic index(MI)of CU group was higher than CM group significantly from 24h-72h after reperfusion(P＜0.01);The apoptosis index(AI)of CU group was lower than CM group at any time point after reperfusion(P＜0.01,P＜0.05);Compared with CM group,the expression of Bcl-2 and NF-_κB mRNA were increased significantly in CU group at any time point after reperfusion.The expression of Bcl-2 and NF-_κB protein in CU group were higher than CM group(P＜0.01,P＜0.05)since 24h after reperfusion and the expression of TNF-αmRNA and protein were reduced significantly in CU group 6h,24h and 48h after reperfusion(P＜0.01,P＜0.05).Conclusion:IRI can retard and impair the liver regenerative function,readily induce hepatocyte apoptosis which may be associated with the continual increased expression of TNF-αinduced by IRI; Curcumin preconditioning can relieve the apoptosis and the mechanism may be associated with inhabiting the continual increased expression of TNF-αand promote the expression of Bcl-2;inhabiting the continual increased expression of TNF-α,promote the expression of NF-_κB, relieve apoptosis may be the key point for the protective effect on liver regeneration of Curcumin.
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