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

[背景与目的]微循环障碍是无心跳供体（NHBD）肝移植（LTx）的主要障碍，其原因是肝窦内皮细胞（SEC）的退化和肝窦狭窄。我们研究了枯否细胞（KC）在NHBD移植物窦状微循环中的作用，重点是NF-κB和细胞因子和类花生酸。我们还研究了KC清除和细胞因子抑制对濒死NHBD 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。实验动物分为4组（n=6）：对照组，不保存肝脏; HB组，冷保存6 h后再灌注肝脏; NHB组，冷保存后再灌注肝脏; KE组，用脂质体包封的二膦酸二氯甲烷预处理大鼠的KC，冷保存6 h后再灌注NHBD肝脏 关于我们 保存。检测NF-κB、细胞因子（TNFα、IL-1β、IL-6）、类花生酸（TXA 2、6-ket-PGF 1 α、LTB 4）、AST、LDH。B）将猪分为5组（n=6）：NHB组，移植来自NHBD的肝移植物; NM组，用甲磺酸萘莫司他（NM）冲洗液冲洗来自NHBD的肝移植物; KC去除组，移植来自NHBD的KC去除的肝移植物; KC去除+NM冲洗组，移植来自NHBD的KC去除的肝移植物，用冲洗的NM冲洗液冲洗;细胞因子抑制组在肝移植前、后分别给予TNFθ、IL 1 β抑制及PGE 1。[结果] A）NHB组NF-κB活性显著高于其他各组。KE组NF-κB活性与对照组相同，细胞因子和类花生酸水平受到抑制，窦状隙结构保存良好。B）在KC去除+NM冲洗和细胞因子抑制组中，5个受体中有4个存活。再灌注后，组织学证明，枯否细胞的消除防止SEC恶化和NM冲洗防止窦狭窄。除NHB组外，其余各组肝组织能荷均恢复正常。[结论] KC活性在NHBD肝移植肝窦微循环障碍中起重要作用，KC活性的调节是NHBD肝移植成功的关键，对防止微循环障碍中SEC恶化和窦缩窄具有重要意义。由抑制KC活性和NM冲洗组成的联合治疗将能够预防濒死NHBD肝移植物中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