Vajdová K, Smreková R, Kukan M, Lutterová M, Wsólová L
Laboratory of Perfused Organs, SCOT, Institute of Preventive and Clinical Medicine, 83301 Bratislava, Slovakia.
Cryobiology. 2000 Sep;41(2):145-52. doi: 10.1006/cryo.2000.2276.
Previous morphological studies failed to show appreciable injury of biliary epithelial cells (BEC) after cold ischemia of rat liver, although recent evidence indicated that BEC integrity and function were impaired in this model. We tested the hypothesis that analysis of bile for enzymes, such as lactate dehydrogenase (LDH), alanine transaminase (ALT), and aspartate transaminase (AST), can be used for assessing cold ischemic injury of BEC. Furthermore, we examined whether biliary gamma-glutamyltransferase (GGT) reflects warm ischemic injury of BEC and whether normothermic reperfusion aggravates the negative effect of cold ischemia on BEC integrity and function. Rat livers were reperfused after different periods of cold or warm ischemia using a blood-free perfusion model. Compared with controls, perfusate LDH, ALT, and AST levels and parameters of hepatocyte function, including hepatocyte tight junction permeability, were not significantly altered by 18-h cold ischemia. On the other hand, 9-h cold ischemia markedly increased biliary LDH, ALT, and AST levels. However, only LDH release into the bile was strongly dependent on the time of cold storage. Biliary GGT, LDH, and glucose levels decreased during the reperfusion period following 18-h cold ischemia. The results suggest that biliary LDH can be used for assessing injury of BEC in cold-preserved livers and that normothermic reperfusion does not aggravate preservation-induced injury of BEC after cold ischemic storage.
以往的形态学研究未能显示大鼠肝脏冷缺血后胆管上皮细胞(BEC)有明显损伤,尽管最近有证据表明在该模型中BEC的完整性和功能受损。我们验证了这样一个假设,即分析胆汁中的乳酸脱氢酶(LDH)、丙氨酸转氨酶(ALT)和天冬氨酸转氨酶(AST)等酶,可用于评估BEC的冷缺血损伤。此外,我们还研究了胆汁γ-谷氨酰转移酶(GGT)是否反映BEC的热缺血损伤,以及常温再灌注是否会加重冷缺血对BEC完整性和功能的负面影响。使用无血灌注模型,对大鼠肝脏进行不同时长的冷缺血或热缺血后再灌注。与对照组相比,18小时的冷缺血并未显著改变灌注液中LDH、ALT和AST的水平以及肝细胞功能参数,包括肝细胞紧密连接通透性。另一方面,9小时的冷缺血显著增加了胆汁中LDH、ALT和AST的水平。然而,只有LDH释放到胆汁中强烈依赖于冷藏时间。在18小时冷缺血后的再灌注期,胆汁GGT、LDH和葡萄糖水平下降。结果表明,胆汁LDH可用于评估冷保存肝脏中BEC的损伤,并且常温再灌注不会加重冷缺血保存后BEC的保存诱导损伤。