Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden.
J Surg Res. 2012 Dec;178(2):959-67. doi: 10.1016/j.jss.2012.06.031. Epub 2012 Jul 6.
Prolonged cold ischemia is frequently associated with a greater risk of delayed graft function and enhanced graft failure. We hypothesized that media, combining a high oxygen-dissolving capacity with specific qualities of organ preservation solutions, would be more efficient in reducing immediate ischemia-reperfusion injury from organs stored long term compared with standard preservation media.
Kidneys retrieved from brain-dead pigs were flushed using either cold histidine-tryptophan-ketoglutarate (HTK) or oxygen-precharged emulsion composed of 75% HTK and 25% perfluorohexyloctane. After 18 h of cold ischemia the kidneys were transplanted into allogeneic recipients and assessed for adenosine triphosphate content, morphology, and expression of genes related to hypoxia, environmental stress, inflammation, and apoptosis.
Compared with HTK-flushed kidneys, organs preserved using oxygen-precharged HTK-perfluorohexyloctane emulsion had increased elevated adenosine triphosphate content and a significantly lower gene expression of hypoxia inducible factor-1α, vascular endothelial growth factor, interleukin-1α, tumor necrosis factor-α, interferon-α, JNK-1, p38, cytochrome-c, Bax, caspase-8, and caspase-3 at all time points assessed. In contrast, the mRNA expression of Bcl-2 was significantly increased.
The present study has demonstrated that in brain-dead pigs the perfusion of kidneys with oxygen-precharged HTK-perfluorohexyloctane emulsion results in significantly reduced inflammation, hypoxic injury, and apoptosis and cellular integrity and energy content are well maintained. Histologic examination revealed less tubular, vascular, and glomerular changes in the emulsion-perfused tissue compared with the HTK-perfused counterparts. The concept of perfusing organs with oxygen-precharged emulsion based on organ preservation media represents an efficient alternative for improved organ preservation.
长时间的冷缺血常伴随着延迟移植物功能和增强移植物衰竭的风险。我们假设，与标准保存介质相比，结合高氧气溶解能力和器官保存溶液特定特性的介质，在减少长期储存器官的即时缺血再灌注损伤方面将更加有效。
使用冷组氨酸-色氨酸-酮戊二酸（HTK）或由 75% HTK 和 25%全氟己烷组成的充氧乳剂冲洗从脑死亡猪中取出的肾脏。冷缺血 18 小时后，将肾脏移植到同种异体受体中，并评估三磷酸腺苷含量、形态以及与缺氧、环境应激、炎症和细胞凋亡相关的基因表达。
与 HTK 冲洗的肾脏相比，使用充氧 HTK-全氟己烷乳剂保存的器官具有更高的三磷酸腺苷含量，并且在所有评估时间点缺氧诱导因子-1α、血管内皮生长因子、白细胞介素-1α、肿瘤坏死因子-α、干扰素-α、JNK-1、p38、细胞色素-c、Bax、caspase-8 和 caspase-3 的基因表达显著降低。相反，Bcl-2 的 mRNA 表达显著增加。
本研究表明，在脑死亡猪中，用充氧 HTK-全氟己烷乳剂灌注肾脏可显著减少炎症、缺氧损伤和细胞凋亡，并且细胞完整性和能量含量得到很好的维持。组织学检查显示，与 HTK 灌注的对应物相比，乳剂灌注组织的肾小管、血管和肾小球变化较小。基于器官保存介质用充氧乳剂灌注器官的概念代表了改善器官保存的有效替代方案。
