Effect of Nitric Oxide on ex vivo normothermic liver perfusion with purely artificial products using artiicial blood.

背景资料。我们尝试开发了体外肝脏器官培养系统，我们之前已经展示了使用人工产品的常温灌流系统的开发。在这项研究中，我们研究了iNOS抑制剂或NO供体在常温灌流系统中使用人工血液的效果。材料和方法。取自一头体重15公斤的母猪的肝移植物，按常规方法采集。灌注液由人工血(Neo红细胞)、L-15培养液、蒸馏水、牛血清白蛋白、碳酸氢钠、氢氧化钠、氯化钾、人普通胰岛素、50%葡萄糖溶液和己米松组成。将该含氧灌流液以300ml/min的速度通过门静脉灌流离体肝6h。结果：与对照组相比，iNOS抑制剂治疗组大鼠血清AST、LDH水平明显降低，氧耗水平明显提高。组织学结果显示，iNOS抑制剂治疗组大鼠脑内3h内坏死区明显少于对照组。灌流液中不加供体可提高Fischer Afs比率，降低丝氨酸、蛋氨酸等细胞毒性氨基酸。结论。选择性诱导型一氧化氮合酶抑制剂可抑制离体肝损伤。无供体给药有助于改善氨基酸代谢。

Background. We tried to development of ex vivo hepatic organ culture system, and we have previously demonstrated that development of a normothermic perfusion system that employed artificial products. In this study, we investigated the effects of administration of iNOS inhibitor or NO donor in a normothermic perfusion system with an artificial perfusate using artificial blood.Materials and Methods. A liver graft from a female pig weighting 15 kg was harvested in the usual manner. The Perfusion solution consisted of artificial blood (Neo Red Cell), L-15 medium, distilled water, bovine serum albumin, NaHCO3, NaOH, KC1, human regular insulin, 50% glucose solution, and Hexamethasone. The isolated liver was perfused with this oxygenated perfusate through the portal vein at a rate of 300 ml/min for 6 hours. Selective iNOS inhibitor or NO donor was administration in this perfusion system.Results., Serum contents of AST and LDH level in iNOS inhibitor treatment group markedly dropped and improve oxygen consumption level compared with control group. Histological findings showed that few necrotic areas in 3 hour after perfusion in iNOS inhibitor treatment group compared with control group. NO donor administaration in the perfusion solution improved FischerAfs ratio and decrease cytotoxic amino acid such as serine and methionine. Conclusions. A selective iNOS inhibitor inhibited isolated liver injury. NO donor administration contributed to improvement of amino acid metabolism.