Brass C A, Roberts T G
Department of Medicine, University of Pennsylvania School of Medicine, Philadelphia.
Gastroenterology. 1995 Apr;108(4):1167-75. doi: 10.1016/0016-5085(95)90216-3.
BACKGROUND/AIMS: Free radicals are important mediators of reperfusion injury; however, the mechanism(s) of oxyradical production after liver reimplantation are not well understood. A model of cold storage and reperfusion using low-level chemiluminescence to directly measure oxyradical production during reperfusion was developed.
Rat livers were harvested and stored at 4 degrees C in University of Wisconsin cold-storage solution or Euro-Collins solution for 0-48 hours and then flushed and reperfused with warm oxygenated (37 degrees C) Krebs-Henseleit buffer. Liver chemiluminescence was measured using a sensitive photomultiplier tube, and hepatocellular injury was assessed by measuring aspartate aminotransferase release into the perfusate.
Chemiluminescence reached a maximum within 5 minutes of reperfusion and then decreased to a baseline within 30 minutes. There was a marked increase in chemiluminescence after only a short period of storage in University of Wisconsin cold-storage solution. Chemiluminescence decreased with longer periods of storage but steadily increased again after 16 hours of storage. Chemiluminescence after 22 hours of storage, but not after 3 hours of storage, was decreased by pretreatment with the Kupffer cell inactivator gadolinium chloride.
The data suggest two mechanisms of oxyradical production during cold storage and reperfusion of the rat liver. The later phase seems to be Kupffer cell dependent.
背景/目的：自由基是再灌注损伤的重要介质；然而，肝脏再植入后氧自由基产生的机制尚未完全明确。我们建立了一种利用低水平化学发光直接测量再灌注期间氧自由基产生的冷保存和再灌注模型。
取大鼠肝脏，置于威斯康星大学冷保存液或欧洲柯林斯液中于4℃保存0 - 48小时，然后用温热的含氧（37℃）克雷布斯 - 亨泽莱特缓冲液冲洗并再灌注。使用灵敏的光电倍增管测量肝脏化学发光，并通过测量灌注液中天冬氨酸转氨酶的释放来评估肝细胞损伤。
化学发光在再灌注后5分钟内达到最大值，然后在30分钟内降至基线。在威斯康星大学冷保存液中仅短暂保存后，化学发光显著增加。化学发光随保存时间延长而降低，但在保存16小时后又再次稳步增加。用库普弗细胞灭活剂氯化钆预处理可降低保存22小时后的化学发光，但不能降低保存3小时后的化学发光。
数据表明大鼠肝脏冷保存和再灌注期间氧自由基产生有两种机制。后期似乎依赖于库普弗细胞。
