Kebis Anton, Kukan Marián, Grancic Peter, Jakubovský Ján
Laboratory of Perfused Organs, Slovak Center for Organ Transplantation, Slovak Medical University, Limbová 12, 83303 Bratislava, Slovakia.
J Zhejiang Univ Sci B. 2007 May;8(5):289-95. doi: 10.1631/jzus.2007.B0289.
BACKGROUND/AIM: Currently, the liver is cold-preserved at 0 approximately 4 degrees C for experimental and clinical purposes. Here, we investigated whether milder hypothermia during the initial phase of the preservation period was beneficial for liver viability upon reperfusion.
In the first set of experiments, rat livers were preserved either conventionally in clinically used histidine-trypthopan-ketoglutarate (HTK) solution (Group A: 45 min and Group B: 24 h) or by slow cooling HTK solution (from 13 degrees C to 3 degrees C) during the initial 45 min of preservation (Group C: 24 h). In the second set of experiments, additional groups of livers were evaluated: Group BB--preservation according to Group B and Group CC--preservation according to Group C. Further, some livers were preserved at 13 degrees C for 24 h. Livers were then reperfused using a blood-free perfusion model.
Bile production was approximately 2-fold greater in Group C compared to Group B. Alanine transaminase (ALT) and aspartate transaminase (AST) release into perfusate were 2 approximately 3-fold higher in Group B compared to Group C. No significant differences were found in ALT and AST release between Group C and Group A. Livers in Group CC compared to Group BB exhibited significantly lower portal resistance, greater oxygen consumption and bromosulfophthalein excretion into bile and lower lactate dehydrogenase (LDH) release into perfusate. Histological evaluation of tissue sections in Group BB showed parenchymal dystrophy of hepatocytes, while dystrophy of hepatocytes was absent in Group CC. Livers preserved at 13 degrees C for 24 h exhibited severe ischemic injury.
These results suggest that the conventional way of liver preservation is not suitable at least for rat livers and that slow cooling of HTK solution during the initial phase of cold storage can improve liver viability during reperfusion.
背景/目的:目前,为了实验和临床目的,肝脏在0至4摄氏度下进行冷保存。在此,我们研究了在保存期初始阶段采用较温和的低温是否有利于肝脏再灌注后的存活能力。
在第一组实验中,大鼠肝脏要么在临床上使用的组氨酸 - 色氨酸 - 酮戊二酸(HTK)溶液中常规保存(A组:45分钟和B组:24小时),要么在保存的最初45分钟内通过缓慢冷却HTK溶液(从13摄氏度降至3摄氏度)进行保存(C组:24小时)。在第二组实验中,对另外几组肝脏进行了评估:BB组 - 按照B组的方式保存,CC组 - 按照C组的方式保存。此外,一些肝脏在13摄氏度下保存24小时。然后使用无血灌注模型对肝脏进行再灌注。
与B组相比,C组的胆汁生成量大约高2倍。与C组相比,B组灌注液中丙氨酸转氨酶(ALT)和天冬氨酸转氨酶(AST)的释放量高约2至3倍。C组和A组之间的ALT和AST释放量没有显著差异。与BB组相比,CC组的肝脏表现出明显较低的门静脉阻力、较高的耗氧量、胆汁中溴磺酞钠排泄量以及灌注液中较低的乳酸脱氢酶(LDH)释放量。BB组组织切片的组织学评估显示肝细胞实质营养不良,而CC组不存在肝细胞营养不良。在13摄氏度下保存24小时的肝脏表现出严重的缺血性损伤。
这些结果表明,传统的肝脏保存方式至少不适用于大鼠肝脏,并且在冷保存初始阶段对HTK溶液进行缓慢冷却可提高肝脏再灌注期间的存活能力。