大鼠心脏停搏供体肝移植供肝活性的研究

Study on viability of the liver graft in rat liver transplantation from non heart beating donors

目的 比较不同类型的大鼠心脏停搏供体 (NHBD)肝移植中供肝的活性 ,探讨肝移植应用NHBD供肝的可行性。方法 应用氧气极谱法测定线粒体呼吸控制率 (RCR ) ;应用荧光法测定线粒体质子ATP酶活性 ;动脉血气分析 ;肝功检查 ;测定血清细胞因子白细胞介素 (IL) 1β ,IL 6和肿瘤坏死因子 (TNF) α作为应激标志物指标 ;肝组织形态学检测。结果 线粒体质子ATP酶活性与热缺血时间 (WIT)呈负相关 ;在KCL3 0组RCR值、ATP酶活性和PaO2 测定值显著高于OC3 0组 (P <0 .0 5 ) ;在心脏停搏 3 0min时 ,血清ALT及LDH值两组都显著增高 ,KCL60组ALT和LDH明显增高组间差异有统计学意义 (P <0 .0 5 )。结论 肝移植应用NHBD供肝是可行的 ,心跳缓慢停止的NHBD模型供肝活性明显优于心脏骤停NHBD。

Objective To examine the difference of graft viabilities in these non heart beating donors (NHBD) models with sudden cardiac arrest induced by injection of KCL and agonal NHBD by cessation of respiration.Availability of liver graft from NHBD in liver transplantation was evaluated.Methods According to modality of cardiac arrest,36 male Wistar mice were divided randomly into three groups.In KCL group (sudden cardiac arrest groups),cardiac arrest was induced by potassium chloride (0.3 g/kg) injected intravenously and was confirmed by opening the chest diaphragm).In group OC (Slow cardiac arrest groups),their respiration was stopped by opening the chest wall.In group HB (control groups),the heart beating mice was without warm ischemic).Such as followed assay was made at 30mins and 60 mins after cardiac arrest,respectively.Mitochondrial respiratory control ratio (RCR) and mitochondrial ATPase activity were examined as the index of mitochondrial ATPase activity.Blood gas (ie,pH,PaO 2,and PCO 2) was analyzed.As liver function tests,ALT and LDH were examined.Serum IL 1β,IL 6,and TNF α were also measured as marker of stress.Morphological changes were investigated by light microscopy.Results Mitochondrial proton ATPase activity decreased with time dependently during WIT.Both values of ATPase activity (34.2± 0.04)% and RCR (3.29± 0.26) in the sudden cardiac arrest group were significantly higher than (24.2± 0.04)% and (2.83± 0.43) in the slow cardiac arrest group at 30 minutes ( P < 0.05). But the value of ATPase activity and RCR in mitochondria decreased to almost the same values at 60 minutes in both groups.The blood gas showed that the PaO 2 in the KCL30 group was (9.0± 1.49) kPa and significantly higher than (5.7± 1.36) kPa in the OC group at 30 minutes ( P < 0.05). Blood pH decreased at 30 and 60 minutes,and there was no significant difference between two groups.Values of serum ALT and LDH significantly increased at 30 minutes in both groups and were significantly higher at 60 minutes in the sudden cardiac arrest group.IL 1β and IL 6 were not detected in either group at 30 or 60 minutes.Value of TNF α, (20.3± 22.0) ng/L,peaked at 30 minutes after warm ischemia in the sudden cardiac arrest group.On the other hand,the value, (27.2± 7.4) ng/L,reached maximum at 60 minutes in the slow cardiac arrest group.Histological findings showed that sinusoid was remarkably widened in the sudden cardiac arrest group,and parenchymal hepatocytes showed hydropic change at 60 minutes in both groups.Conclusion Liver transplantation from NHBD is feasible.The viability of liver graft in slow cardiac arrest was significantly superior to that of in the sudden cardiac arrest model.