胆汁中总胆汁酸盐/磷脂比例与大鼠移植肝胆道相对热缺血损伤的相关性  

Correlation between total bile acid/phospholipid ratio and relative warm ischemia injury to the biliary tract in rat liver transplantation

作　　者：任旋磊[1] 赵宏峰[1] 张国伟[1] 王宇[1] 周杰[1] 

机构地区：[1]南方医科大学南方医院肝胆外科,广东省广州市510515

出　　处：《中国组织工程研究与临床康复》2010年第5期818-823,共6页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：广东省科技计划项目(2007B031514004)资助~~

摘　　要：背景:据报道,供肝的热缺血、冷保存时间过长都可导致移植肝产生"毒性胆汁",并进而损伤胆道,然而胆道相对热缺血是否也会引起胆汁成分改变,目前尚无相关报道。目的:建立大鼠自体原位肝移植胆道相对热缺血损伤模型,观察胆道相对热缺血对大鼠移植肝分泌的胆汁中总胆汁酸盐、磷脂浓度的影响及与胆道损伤的相关性。方法:选用SD大鼠32只建立大鼠自体原位肝移植胆道相对热缺血损伤模型。按随机数字表法分为4组(n=8):Ⅰ组为假手术组,只游离肝脏,不进行肝脏冷灌注;Ⅱ、Ⅲ、Ⅳ组的胆道相对热缺血时间分别为0,30,60min,即分别在门静脉开放后0,30和60min时开放肝动脉。采用循环酶法测定术后胆汁中总胆盐浓度,酶比色法测定磷脂浓度。取肝门部胆管,光镜观察其组织学形态,透射电镜观察胆道上皮细胞的超微结构变化,原位末端标记法检测上皮细胞凋亡情况。分析总胆汁酸盐/磷脂比例与胆道损伤的相关性。结果及结论:死亡1只,进入结果分析31只。肝移植胆道相对热缺血可以引起移植肝分泌的胆汁成分改变,表现为总胆汁酸盐/磷脂比例升高,胆汁毒性增加,这种改变与相对热缺血时间呈正相关,相对热缺血时间越长,胆汁成分改变越明显;而且与胆道损伤的病理学改变呈密切正相关,说明胆汁成分改变是相对热缺血导致胆道损伤的重要机制之一。BACKGROUND：Studies have shown that long time of warm ischemia or cold preservation would injury the biliary tract in liver transplantation.However,whether relative warm ischemia（RWI） of biliary tract would result in bile component changes is unclearly.OBJECTIVE：To establish auto-liver transplantation bile ducts RWI models,observe the effects of RWI on the bile salts and phospholipid concentration secreted by the donor liver,and to study the correlation between the total bile salt/phospholipid ratio（TBA/PL ratio） and biliary tract injury.METHODS：A total of 32 SD rats were selected for auto-liver transplantation models with bile ducts RWI,and the rats were randomly divided into 4 groups（n=8）.In Group Ⅰ（sham operation group）,rats only received liver dissociation without any cold reperfusion.The RWI time of Group Ⅱ,Ⅲ,and Ⅳ were 0 minutes,30 minutes and 60 minutes,respectively.The concentration of TBA in bile was measured with enzymatic cycling assay,and PL with enzymic colorimetric.Pathological observation with light microscope and ultrastructural observation with transmission electron microscope were performed on the hilar bile duct.The endothelial cell apoptosis was detected with TUNEL assay.The correlation between TBA/PL ratio and biliary injury was analyzed.RESULTS AND CONCLUSION：One rat died,the other 31 rats were included in the final analysis.RWI could change the composition of bile secreted by donor liver,raise the TBA/PL ratio,and increase the bile toxicity.These changes had a positive correlation to RWI time,and the changes were obviously with time prolonged.In addition,the changes are closely related to the biliary tract injury.This study shows an important mechanism of the biliary tract injury caused by RWI-injury.

关 键 词：胆道相对热缺血损伤 胆汁 器官移植 肝移植 胆汁酸盐 磷脂 

分 类 号：R617[医药卫生—外科学]