| PurposeEstablish a New Zealand rabbit allograft orthotopic kidney transplantation model first,then preserve the rabbit allograft kidney and perform kidney transplantation by two methods of antegrade continuous hypothermic perfusion through renal artery and retrograde continuous hypothermic perfusion through renal vein,after that,observe the creatinine,urine volume,pathology of the allograft kidney tissue,expression levels of malondialdehyde(MDA)and super oxide dismutase(SOD)in the allograft kidney tissue of the two groups,and preliminarily explore the influence of different perfusion methods on the quality of the donor kidney,so as to provide new strategies and ideas for clinical expansion of the use of marginal kidney donors.Methods1.An animal model of allogeneic orthotopic kidney transplantation after thermal ischemia injury was established: donor New Zealand rabbit's left renal artery was blocked for 45 min to form donor renal thermal ischemia injury.Two different methods of antegrade continuous low temperature perfusion and retrograde continuous low temperature perfusion were used for perfusion for 2 hours,and then orthotopic transplantation was performed to the recipient.After transplantation,the recipient's right kidney was excised,and cyclosporine A was used for anti-rejection after surgery,thus completing the experimental model.2.Sixty healthy adult New Zealand male rabbits weighing 2-3kg were purchased from the Animal Laboratory of South China University.Rabbits were divided into two groups by random number table method,anterograde continuous hypothermia perfusion group(group A)and retrograde continuous hypothermia perfusion group(group B)with 30 rabbits in each group,then rabbits were randomly divided into donor and recipient groups in group A and B with 15 rabbits in each group.The levels of serum creatinine and urine volume of recipient New Zealand rabbits in anterograde perfusion group and retrograde perfusion group were compared on the first day,the third day and the fifth day after operation.The pathological changes of HE staining and the expression of MDA and SOD activity in transplanted kidney tissue were observed on the 5th day after operation.Results1.Changes of serum creatinine: In group A(antegrade perfusion group),the serum creatinine values on the first,third and fifth postoperative days were:(398.36±37.28)umol/L,(318.97±18.34)umol/L,(178.01±9.44)umol/L,respectively;In group B(retrograde perfusion group),the serum creatinine values on the first,third and fifth postoperative days were:(417.51±34.28)umol/L,(330.54±20.72)umol/L,(186.21±13.81)umol/L,respectively.There was no significant difference in the postoperative serum creatinine between retrograde perfusion group and anterograde perfusion group(P > 0.05).2.Changes of urine volume:In group A(antegrade perfusion group),the urine creatinine values on the first,third and fifth postoperative days were(74.60±5.55)ml,(93.72±8.89)ml,(137.58±9.79)ml,respectively;the urine creatinine values on the first,third and fifth postoperative days were(71.18±5.79)ml,(88.16±6.84)ml and(130.99±12.11)ml,respectively.There was no significant difference in postoperative urine volume between retrograde perfusion group and anterograde perfusion group(P > 0.05).3.HE staining pathology of transplanted kidney tissue: group A(anterograde perfusion group)and group B(retrograde perfusion group)showed a little microthrombotic substance was formed in glomerulus and peripheral capillaries,and the renal tissue structure was basically clear on the 5th day after operation.The lumen of renal tubules was slightly enlarged.A little congestion,edema and inflammatory cell distribution in the stroma.4.The expression of MDA in transplanted kidney tissue was detected by spectrophotometry: MDA content in group A(anterograde perfusion group)and group B(retrograde perfusion group)were(6.65±0.60)nmol/mgprot and(7.02±0.55)nmol/mgprot respectively on the fifth day after operation.There was no significant difference in MDA content between retrograde perfusion group and anterograde perfusion group after operation,P>0.05,the difference was not statistically significant.5.The expression of SOD activity was detected by spectrophotometry in transplanted kidney tissue: SOD activity in group A(antegrade perfusion group)and group B(retrograde perfusion group)was(369.35±35.37)U/mgprot and(349.13±31.96)U/mgprot respectively on the fifth day after operation.There was no significant difference in SOD activity between retrograde perfusion group and anterograde perfusion group after operation,P>0.05,the difference was not statistically significant.ConclusionsThe retrograde perfusion method not only protects the function of the transplanted kidney as well as the antegrade perfusion method,but also serves as a remedial for lavage when the renal artery is malformed and damaged. |
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