Kuroda Y, Matsumoto S, Fujita H, Tanioka Y, Sakai T, Hamano M, Hiraoka K, Kim Y, Suzuki Y, Ku Y, Saitoh Y
First Department of Surgery, Kobe University School of Medicine, Japan.
Transplantation. 1996 Jan 15;61(1):28-30. doi: 10.1097/00007890-199601150-00007.
We have shown that 24-hr preservation by a two-layer (University of Wisconsin solution [UW]/perfluorochemical [PFC]) cold storage method allows tissue ATP synthesis and makes it possible to resuscitate a canine pancreas subjected to 90 min of warm ischemia. The purpose of this study was to examine whether increasing preservation temperature to 20 degrees C makes it possible to shorten a preservation period for recovery of ischemically damaged pancreas grafts. After 90 min of warm ischemia, canine pancreas grafts were preserved using the two-layer (UW/PFC) method for 1 to 8 hr at 20 degrees C, and then autotransplanted. A K-value of intravenous glucose tolerance test more than 1.0 at 2 weeks after transplantation was considered graft survival. ATP tissue levels were measured by high performance liquid chromatography at the end of preservation. Pancreatic tissue perfusions were measured using an H2 clearance technique after 30 min to 4 hr of reperfusion. Pancreas grafts subjected to 90 min of warm ischemia were not viable (0/5, control group). However, 3- and 5-hr preservations made it possible to recover the ischemically damaged pancreas (3/5 and 5/5, respectively), although 1- and 8-hr preservations were not successful (0/3 and 0/3, respectively). ATP tissue levels in 1-hr-preserved grafts were 2.55 +/- 0.38 mumol/g dry weight and were significantly lower compared with the levels in 5- and 8-hr-preserved grafts, 9.40 +/- 2.09 (P < 0.01) and 7.37 +/- 1.06 (P < 0.01), respectively. On the other hand, pancreatic tissue perfusions in 8-hr-preserved grafts after 2 hr of reperfusion were 28.50 +/- 7.52 ml/100 g/min and were significantly lower than the values in 1- and 5-hr-preserved grafts, 66.0 +/- 11.22 (P < 0.01) and 57.10 +/- 4.40 (P < 0.01), respectively. It was suggested that 1-hr-preservation was not enough to synthesize ATP, which was essential to repair damaged cells, although vascular microcirculation at reperfusion was maintained and 8-hr preservation incurred microcirculatory disturbances, although ATP for repairing damaged cells was synthesized. We conclude that 3- to 5-hr preservation at 20 degrees C by the two-layer (UW/PFC) method accelerates ATP synthesis, which is essential for repairing damaged cells and protects vascular microcirculation. This makes it possible to resuscitate ischemically damaged pancreases faster. This method holds promise for pancreas-kidney transplantation from cardiac arrest donors.
我们已经表明,采用双层(威斯康星大学溶液[UW]/全氟化合物[PFC])冷藏法保存24小时可使组织进行ATP合成,并能够使经历90分钟热缺血的犬胰腺复苏。本研究的目的是检验将保存温度提高到20℃是否能够缩短缺血损伤胰腺移植物恢复所需的保存期。在90分钟热缺血后,犬胰腺移植物采用双层(UW/PFC)法在20℃保存1至8小时,然后进行自体移植。移植后2周静脉葡萄糖耐量试验的K值大于1.0被视为移植物存活。在保存结束时通过高效液相色谱法测量ATP组织水平。在再灌注30分钟至4小时后,使用H2清除技术测量胰腺组织灌注。经历90分钟热缺血的胰腺移植物无活力(0/5,对照组)。然而,3小时和5小时的保存能够使缺血损伤的胰腺恢复(分别为3/5和5/5),尽管1小时和8小时的保存未成功(分别为0/3和0/3)。1小时保存的移植物中ATP组织水平为2.55±0.38μmol/g干重,与5小时和8小时保存的移植物中的水平相比显著降低,分别为9.40±2.09(P<0.01)和7.37±1.06(P<0.01)。另一方面,8小时保存的移植物在再灌注2小时后的胰腺组织灌注为28.50±7.52ml/100g/min,显著低于1小时和5小时保存的移植物中的值,分别为66.0±11.22(P<0.01)和57.10±4.40(P<0.01)。结果表明,1小时的保存不足以合成对修复受损细胞至关重要的ATP,尽管再灌注时维持了血管微循环;而8小时的保存虽然合成了用于修复受损细胞的ATP,但引起了微循环紊乱。我们得出结论,采用双层(UW/PFC)法在20℃保存3至5小时可加速ATP合成,这对修复受损细胞至关重要并能保护血管微循环。这使得能够更快地使缺血损伤的胰腺复苏。该方法有望用于心脏骤停供体的胰肾联合移植。