| Objective Through the establishment of rat kidney warm ischemia model,to Simulate in the DCD donor kidney transplantation,the donor underwent a period of warm ischemia,To explore the protective effect of continuous hypothermic perfusion of urokinase on donor kidney,So as to provide new methods and ideas for clinical improvement of renal function of DCD.Methods(1)60 male inbred SD rats(12-14 weeks)were randomly divided into 3 groups: 30 min,45min,60 min of warm ischemia time in group A,group B and group C respectively,each group of 20.Left renal artery and vein were clipped simultaneously until the setting time.Left kidney was resected for examination.The pathologic change of the kidney were observed by HE,Masson staining.(2)Both kidneys of 30 male inbred SD rats(12-14 weeks)were randomly divided into 2 groups according to coin-tossing method: control group were treated with HCA liquid and normal saline,experimental group were treated with HCA liquid and urokinase.Abdominal aorta were clipped between the left and right renal artery for 45 min to simulate the process of warm ischemia,the inferior vena cava were clipped between the left and right renal vein simultaneously.Then the left kidney was treated by cold perfusion liquid through the abdominal aorta below.After the perfusion was finished,Left kidney were resected.Then the clips were released,Abdominal aorta were clipped above the right renal artery again,inferior vena cava were clipped above the right renal vein simultaneously,the right kidney was perfused as the same way,After the perfusion was finished,the right kidney was resected for examination too.The histological change of the kidney,thrombomodulin and NGAL expressing level were observed by HE,Masson staining and immunohistochemistry.Results(1)Under light-microscopy,rare thrombosis in glomerulus and nothing abnormal in renal tubules were observed in WI 30 min,while severer tubular performance such as vacuolation of renal tubular epithelial cells,a large amount of micro thrombi in glomerulus were observed in WI 45 min and WI 60 min.the histological damage scores were significantly lower compared with the WI 45 min and WI 60min(P<0.05),while in WI 60 min group,the scores were higher than WI 45min(P<0.05).A lot of micro thrombi in glomerulus and severe damage in renal tubules were observed in control group,while thrombosis and abnormal renal tubules were rarely seen in experimental group.The histological damage scores were significantly lower compared with the control group(P<0.05).(2)Compared with the results of control group in which the expression of TM in renal tissue were darker.The expression level of TM in control group was greatly higher than experimental group(P<0.05).(3)NGAL was observed in the proximal tubules in renal tissue in both group,Its expression expanded and strengthened in control group compared with experimental group.The expression level of NGAL in control group was significantly higher than experimental group(P<0.05).Conclusion(1)Thrombosis and kidney injury occurred when over 45 min of warm ischemia.Along with the extension of warm ischemia time,pathological changes developed towards irreversible direction,the injury became very severe and irreversible when over 60 min of warm ischemia.(2)There were very obvious micro thrombi after warm ischemia of 45 min,which caused the kidney structure destroyed and the kidney function decreased.The micro thrombi were dissolved after the treatment of continuous hypothermic perfusion of urokinase,which could help the recovery of the structure and function of thekidney. |
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