MECHANISMS AND PREVENTION OF GRAFT INJURY OF THE LIVER GRAFT IN NON-HEART-BEATING DONOR.

[背景与目的]微循环障碍是非心脏供体肝移植（NHBD）的主要障碍，微循环障碍是由窦内皮细胞（SEC）恶化和窦狭窄引起的。我们研究了Kupffer细胞（KC）在NHBD移植物窦状微循环中的作用，重点是NF-κB和细胞因子和类二十烷酸。我们还研究了KC消除和细胞因子抑制对agonal NHBD中LTx的影响。[材料/方法]A)分离Wistar大鼠肝脏，用Krebs-Henseleit碳酸氢盐缓冲液灌注60分钟。将动物分为4组（n=6）：对照组，肝脏灌注不保存；HB组，低温保存6小时后灌注肝脏；NHB组，NHBD肝冷保存后灌注；KE组大鼠经脂质体包封二氯亚甲基二膦酸钠预处理以消除kc。检测NF-κB、细胞因子（TNFα、IL-1β、IL-6）、类二十烷酸（TXA2、6-ket-PGF1α、LTB4）、AST和LDH。B)将猪分为5组（n=6）： NHB组，移植NHBD的肝移植；NM组，NHBD接枝用甲磺酸那莫他酯（NM）冲洗液冲洗；KC *化组，KC消去的NHBD移植物移植；KC消除+NM漂洗组，KC消除的NHBD接枝用漂洗NM漂洗液移植；细胞因子抑制组，抑制TNFθ和il - 1β，并在NHBD肝Tx前后给予pge1。[结果]A) NHB组NF-κB活性显著高于其他各组。KE组抑制NF-κB活性与对照组相同，抑制细胞因子和类二十烷素水平，保持良好的正弦结构。B)在KC消除+NM冲洗组和细胞因子抑制组中，5个受者中有4个存活。再灌注后，组织学显示，消除Kupffer细胞可防止SEC恶化，NM冲洗可防止窦狭窄。除NHB组外，其余各组肝组织能量电荷均恢复正常。[结论]KC活性在NHBD对Tx正弦微循环的干扰中起重要作用。KC的调节是NHBD肝移植成功的关键，对防止SEC恶化和窦狭窄的微循环障碍具有重要意义。由抑制KC活性和NM冲洗组成的联合治疗将能够预防非外源性NHBD肝移植的原发性移植物无功能。少

[Background and Purpose] Microcirculatory disturbance is a main obstacle to liver transplantation (LTx) from non-heart beating donors (NHBD) and it is attributed to a deterioration of sinusoidal endothelial cells (SEC) and sinusoidal narrowing. We examined roles of Kupffer cells (KC) in the sinusoidal microcirculation of the graft of NHBD, focusing on NF-κB and cytokines and eicosanoids. We also investigated effects of KC elimination and cytokine suppression on LTx from agonal NHBD, [Materials/Methods] A) Livers of Wistar rat were isolated and perfused for 60 minutes with Krebs-Henseleit bicarbonate buffer. Animals were divided into four groups as follows (n=6) : control group, liver was perfused without preservation ; HB group, livers were perfused after 6 hr cold preservation ; NHB group, liver from NHBD was perfused after cold preservation ; KE group, rats were pretreated with liposome-encapsulated dichloromethlene diphoshonate to eliminate KC.Liver from NHBD was perfused after cold … More preservation. NF-κB, cytokines (TNFα, IL-1β, IL-6), eicosanoids (TXA2, 6-ket-PGF1α, LTB4), AST and LDH were measured. B) Pigs were allocated to five groups (n=6) : NHB group, liver graft from NHBD was transplanted ; NM group, graft from NHBD was rinsed with nafamostat mesilate (NM) rinse solution ; KC *inated group, KC eliminated graft from NHBD was transplanted ; KC eliminated+NM rinse group, KC eliminated graft from NHBD was transplanted with rinsing NM rinse solution ; cytokine suppression group, TNFθ and IL1β were suppressed and PGE1was administered before and after liver Tx from NHBD.[Results] A) NF-κB activity of the NHB group was significantly higher than those of the other groups. In KE group, NF-κB activity was suppressed as same level as in control group, In KE group, cytokine and eicosanoid levels were suppressed and sinusoidal structures were preserved well. B) Both in KC eliminated +NM rinse and cytokine suppression groups, four of five recipients survived. After reperfusion, it was histologically demonstrated that elimination of Kupffer cells prevented SEC deterioration and NM rinse prevented sinusoidal narrowing. The energy charge of liver tissue recovered in all groups except NHB group. [Conclusions] KC activities have an important roll in the disturbance of sinusoidal microcirculation in Tx from NHBD.Modulation of KC is a key for success in liver transplantation from NHBD and it is important to prevent microcirculatory disturbance of SEC deterioration and sinusoidal narrowing. Combination therapy consisting of suppression of KC activities and NM rinse will be able to prevent primary graft non-function in liver grafts from agonal NHBD. Less