Drognitz Oliver, Liu Xuemei, Obermaier Robert, Neeff Hannes, von Dobschuetz Ernst, Hopt Ulrich Theodor, Benz Stefan
Department of General and Digestive Surgery, University of Freiburg, Hugstetter Strasse 55, 79106, Freiburg im-Bresigau, Germany.
Transpl Int. 2004 Jul;17(6):317-24. doi: 10.1007/s00147-004-0704-9. Epub 2004 Jun 24.
Brief periods of warm ischemia and subsequent short reperfusion before either long-term cold or warm ischemic insult (ischemic preconditioning, IPC) have proven to ameliorate ischemia/reperfusion (I/R) injury in various organs, such as the liver and lung. The aim of this study was to examine the effect of IPC on pancreatic cell apoptosis and microcirculatory impairments in experimental pancreas transplantation. Male Lewis rats served as donors and recipients of heterotopic syngeneic pancreaticoduodenal transplantation. Recipient animals were divided into two experimental groups: group Tx (n=7) received grafts without IPC, group Tx&IPC received grafts with IPC. Animals that had not undergone transplantation but whose pancreata had been exteriorized served as controls (n=5). All pancreatic grafts were preserved in University of Wisconsin solution for 6 h at 4 degrees C. IPC was induced by interruption of the arterial blood flow for 10 min followed by 10 min of reperfusion. One and two hours after reperfusion, graft microcirculation was assessed by means of intravital microscopy (IVM). Rats were immediately killed after the second measurement and DNA breaks of acinar cells were detected by in situ terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate digoxigenin nick end-labelling (TUNEL) assay and gel electrophoresis (laddering). The apoptotic index (AI) was defined as the number of apoptotic cells per high-power field. Analysis of both groups of transplanted grafts showed a significant decrease in functional capillary density (FCD) and a significant increase in leukocyte sticking to postcapillary venules (LAV) at 1 h and 2 h of reperfusion, compared with animals that had not undergone transplantation ( P<0.01). In parallel, AI was significantly increased in transplanted grafts compared to the controls ( P<0.01). Grafts subjected to IPC showed no significant differences, neither for FCD nor LAV, at both time points if compared with grafts of group Tx. However, IPC resulted in a significant increase in AI ( P<0.05). We can conclude that IPC has no effect on pancreatic microcirculation but enhances acinar cell apoptosis in experimental pancreas transplantation. These results indicate that IPC might increase I/R injury after pancreatic cold ischemia.
在长期冷或热缺血损伤（缺血预处理，IPC）之前，短暂的热缺血期及随后的短期再灌注已被证明可改善肝脏和肺等多种器官的缺血/再灌注（I/R）损伤。本研究的目的是检测IPC对实验性胰腺移植中胰腺细胞凋亡和微循环障碍的影响。雄性Lewis大鼠作为异位同基因胰十二指肠移植的供体和受体。受体动物分为两个实验组：Tx组（n = 7）接受未进行IPC的移植物，Tx&IPC组接受进行了IPC的移植物。未进行移植但胰腺已外露的动物作为对照（n = 5）。所有胰腺移植物在4℃下于威斯康星大学溶液中保存6小时。通过阻断动脉血流10分钟，随后再灌注10分钟来诱导IPC。再灌注1小时和2小时后，通过活体显微镜检查（IVM）评估移植物微循环。在第二次测量后立即处死大鼠，通过原位末端脱氧核苷酸转移酶介导的脱氧尿苷三磷酸地高辛标记（TUNEL）测定和凝胶电泳（梯状条带）检测腺泡细胞的DNA断裂。凋亡指数（AI）定义为每高倍视野下凋亡细胞的数量。对两组移植移植物的分析显示，与未进行移植的动物相比，再灌注1小时和2小时时，功能性毛细血管密度（FCD）显著降低，白细胞黏附于毛细血管后微静脉（LAV）显著增加（P < 0.01）。同时，与对照组相比，移植移植物中的AI显著增加（P < 0.01）。与Tx组的移植物相比，接受IPC的移植物在两个时间点的FCD和LAV均无显著差异。然而，IPC导致AI显著增加（P < 0.05）。我们可以得出结论，在实验性胰腺移植中，IPC对胰腺微循环无影响，但会增强腺泡细胞凋亡。这些结果表明，IPC可能会增加胰腺冷缺血后的I/R损伤。
