Impact Of Different Time Points Of Donor Liver Removal On The Ischemia-reperfusion Injury Of The Bile Duet In DCD Rat Liver Transplantation After Cardiac Arrest And Cardiopulmonary Resuscitation

Chapter1 Establishment and evaluation of the rat model of cardiac arrest and cardiopulmonary resuscitationObjective To establish a stable rat model of cardiac arrest and cardiopulmonary resuscitation(CPR).Methods The donor rat's abdominal aorta was clipped for 5 minutes to simulate the cardiac arrest and CPR.The MAP?PaCO2 and PaO2 of the controlled group and the experimental group were examined.Results The MAP of the experimental group dropped to 0 after the clipping,witch was(<10mmHg)significantly lower than that of the controlled group(P<0.05).After relasing the clip of the abdominal aorta the MAP of the experimental group was more than 60mmHg,which was the same as that of the controlled group(P>0.05).The Pa02 of the experimental group dropped gradually after the clipping,and it was back to normal when 10 minutes after relasing the clip of the abdominal aorta,which was the same as that of the controlled group(P>0.05).The PaC02 of the experimental group increased gradually after the clipping,and it was back to normal when 10 minutes after relasing the clip of the abdominal aorta,which was the same as that of the controlled group(P>0.05).The serum levels of ALT of the experimental group were significantly higher than that of the controlled group(P<0.05).Conclusion The established rat model of cardiac arrest and cardiopulmonary resuscitation(CPR)was stable,reliable and reproducible,which could simulate the cardiac arrest and CPR of the donor before liver transplantation.Chapter2 Establishment of the rat model of orthotopic liver transplantation after cardiac arrest and cardiopulmonary resuscitationObjective To establish a stable rat model of orthotopic liver transplantation(OLT)after cardiac arrest and cardiopulmonary resuscitation(CPR).Methods The OLT model was established using Kammada two-cuff technique with little modification.The donor rat's abdominal aorta was clipped for 5 minutes to simulate the cardiac arrest and CPR before transplantation.The serum levels of ALT of the controlled group and the experimental group were examined before transplantation and 6 hours?1 day?3 days?7 days after operation.Results Of the total 30 cases of surgery,28 cases were successful,and the success rate of the model was 93%(28/30).The donor and recipient operation time was(49±4)minutes and(54±5)minutes,respectively.The anhepatic phase was(17±2)minutes.The serum levels of ALT of the experimental group and the controled both increased after the operation,reached the highest value at 6 hours after the operation,and then were back to normal when 7 days after the operation.The serum levels of ALT of the experimental group were significantly higher than that of the controlled group(P<0.05).Conclusion Based on the Kammada two-cuff technique,the rat model of OLT after cardiac arrest and CPR is stable,reliable and reproducible.The model simulated the cardiac arrest and CPR of the donor before liver transplantation,providing a new experimental method of studying the liver transplantation of this kind of donors.Chapter3 Impact of different time points of donor liver removal on the ischemia-reperfusion injury of the bile duet in DCD rat liver transplantation after cardiac arrest and cardiopulmonary resuscitationObjective To explore Impact of different time points of doner liver removal on the ischemia-reperfasion injury of the bile duet in DCD rat liver transplantation after cardiac arrest and cardiopulmonary resuscitation.Methods Based on the established rat model of OLT after cardiac arrest and CPR in experiment 2,the rats were divided into 5 groups according to the different time points of doner liver removal after relasing the clip of the abdominal aorta.Group LR2h:remove the doner's liver 2 hours after relasing the clip of the abdominal aorta;Group LR4h:remove the doner's liver4 hours after relasing the clip of the abdominal aorta;Group LR6h:remove the doner's liver 6 hours after relasing the clip of the abdominal aorta;Group LR8h:remove the doner's liver 8 hours after relasing the clip of the abdominal aorta;Group LR10h:remove the doner's liver 10 hours after relasing the clip of the abdominal aorta.At the same time,the sham group and the controlled group were established.The serum levels of ALT?GGT and ITBL of each group 6 hours,1 day,3 days,7 days after operation were examined.SOD enzymatic activity of the rat bile duet and histological analysis of the rat bile duet of each group after the operation were examined.TUNEL was performed to detect the apoptosis of the bile duet cells.Results The serum levels of ALT,GGT and ITBL of the experimental groups and the controlled group both increased after the operation,ALT levels of the experimental groups reached the highest value at 6 hours after the operation,and then were back to normal when 3days after the operation,while the serum levels of GGT and ITBL of the experimental groups were still increasing at 6 hours after the operation,reached the highest value at 3 days after the operation,and then were back to normal when 7 days after the operation.The serum levels of ALT,GGT and ITBL of the LR6h group were significantly higher than that of other groups(P<0.05).SOD enzymatic activity of the rat bile duet of the experimental groups and the controlled group both decreased after the operation,reached the lowest value at 1 day after the operation,and then were back to normal when 7days after the operation.SOD enzymatic activity of the rat bile duet of the LR6h group were significantly lower than that of other groups(P<0.05).HE stained paraffin sections of the rat bile duet of each group afterthe operation showed that pathological changes of the LR6h group were more serious than that of other groups(P<0.05=.The TUNEL method showed that the apoptosis index of the bile duet cells of the LR6h group were much higher than that of other experimental groups(P<0.05).Conclusion The ischemia-reperfusion injury of the bile duet was most severe when remove the liver 6 hours after cardiac arrest and cardiopulmonary resuscitation in DCD rat liver transplantation,and we should not remove the liver at this time.