| Objecttve: To study the effect of CyclosporinA preconditioning on rat donors liver during cold preservation and the influence after orthotopic liver transplantation. (OLT)Materials and Methods: A total of 130 inbred male SD rats, weighing 210± 20g, orthotopic liver transplantation was performed according to the Kamadas cuff tcchnique with modifications .The survival rates in 7 days in different groups were observed.Rats were divided into two groups randomly,Group A (control group): Donors were perfused with 4 °C HTK and stored in the same solusion; Group B (CyclosporinA preconditioning group): donors were perfused with 4°C HTK solution with luM CsA preconditioning.and stored in it. The livers of two groups had been all preservated for 12 hours before orthotopic liver transplantation. Eight time points were predetermined as oh,3h, 6h,12h during preservation and 12h,24h,48h,72h,168h (7days) postoperation. 5-10 rats were obersved everytime.Rats were Sacrificed at each time-point and the liver specimens and blood samples were collected. The hepatocyte injure was evaluated by the level of sperm ALT,AST,LDH.The RCR, P/O and ATP were used to measure the mitochondrial respiration function of hepatocyte.Morphometry was observed under light microscope and transmission electron microscope(TEM). Additionally, the incidence of apopotosis in liver tissue were measured respectively by TUNEL method, The expression level of Bcl-2 in the liver were assessed by immunohistochemistry. All datas are given X± S.E.M, The data were analyzed with spss10.0 , Statistical comparisons were made using t test or ANOVA ofanalysis of variance. AP value<0.05 was considered statistically significant.Results:? The level of sperm ALT,AST and LDH in the two groups at different time-piont:Biochemical data showed that the level of ALT, AST,LDH was significantely higher in group B that were 65.57±7.89, 34.71 ±6.26 and 473.85±285.66 than in group A that were 94.86±27.19 ,58.29±22.27 and 681.38±160.32 which had been preservated for 12 hours, P<0.05.The three parameters of two groups were no statistically significant during preservation 0h,3h ,6h. P>0.05. The level of ALT, ,AST and LDH was significantely lower in group B at the following time point: 12 hours ,24 hours ,48 hours .72 hours, than group A, the data was 788±94.43, 568.2±55.62, 916.6±53.26; 498.8±77.83, 259±71.16, 687.8±43.52; 398.4±35.22, 197.2±32 respectively PO.05. The level of LDH in the two groups were statistically significant during postoperation 168 h. PO.05.(D The prarameters of RCR,P/O in the two groups in different time-piont: The parameters of RCR and P/O of mitochondrial respiraton function in group B after preservation 12h were 3.20±0.24,that in group A were 3.05 + 0.24. Group B was higher than group A , (PO.05). The level of RCR and P/O postoperation for 12h ,24h ,168h in group A were 2.37±0.13,1.86±0.44,3.47±0.25. those in group B were 2.61 ±0.11,2.40±0.24,3.61±0.12 .Group B were significantly higher than gronp A ,(PO.05).(3) The level of ATP in the two groups1 liver tissue in different time-piont: The level of ATP in gronp B which had been preservated for 12h and postoperation,forl 2h,24h,48h,72h, 168hwerel. 19±0.04,0.85±0.10,1.20±0.08,1.22±0 .08,1.25±0.05,1.43±0.04.Those in the group A were 1.05±0.10, 0.56±0.17, 1.05±0.03, 1.09±0.06, 1.16±0.04, 1.29±0.09. Those in group B were much more different than in group A,PO.05.(4) The morphological changes and observation by TEM of two groups' liver: The morphological changes of the livers were more severe in group A than in group B,Prticularly the changes occurred during preservation for 12 hours.The granulocyte inflitrated significantly.The hepatocyte occurred denegeration,spot orbrige shape necrosis during postoperation 12h,24h,48h. The observation of TEM in group A : The mitochondry occurred edma ,the spine breaked ,the hepatocyte nuclear chromatin was disintegrated.The changes in group B were lighter and appearance of the cell nuclear fission is obvious.(5) Result of TUNEL determination:The number of cell apoptosis of liver tissue in two groups increased during preservation for 12 hours. The apoptosis AI was significantly higher in growp A during preservation for 12h and postoperation for half an hour ,12h,24h,48h,72h,168h than in gronp B, P<0.05.(6) Expression of Bcl-2 in the two group liver tissue in different time-piont: The expression of Bcl-2 was significantly higher in group B during preservation for 12h and postoperation for half an hour ,12h,24h,48h,72h,168h than gronp A, P |
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