摘要目的 探讨过氧化物酶体增殖物活化受体-γ(PPAR-γ)配体罗格列酮在原位肝移植胆道缺血-再灌注损伤中作用的分子机制.方法 40只SD大鼠随机(随机数字法)分成假手术组(SO组)、缺血-再灌注组(I/R组)、罗格列酮组(ROS组)和GW9662组,每组10只.通过改良"双袖套法"建立大鼠自体原位肝移植胆道缺血-再灌注模型,通过肝脏及胆管组织病理学变化、血生化指标检测评价模型建立是否成功.SO组行自体原位肝移植术,I/R行建模缺血-再灌注,ROS在缺血-再灌注后以罗格列酮0.3mg/kg经门静脉注射,GW9662在ROS基础上10 min后经门静脉以0.3mg/kg注射GW9662,各组均于实验后4h取部分肝脏和胆管组织用于免疫组化检测;右心室穿刺抽血采用ELISA法测定细胞因子含量.采用方差分析进行统计学处理.结果 组织中细胞因子IL-1β,TNF-α,IL-6活性主要表达在肝细胞及胆管细胞胞浆中,转录因子NF-κB则在胞浆及胞核中均有表达;I/R组及GW9662组上述蛋白表达升高,和SO组以及ROS组比较明显增高(P＜0.05).血清中I/R组大鼠IL-1β,TNF-α及IL-6同步升高,较SO组明显增高(P＜0.05);罗格列酮干预后血清中IL-1β,TNF-α及IL-6和SO组比较无明显变化(P＞0.05);给予罗格列酮阻滞剂GW9662后,IL-1β,TNF-α及IL-6的含量和SO组比较则明显增高(P＜0.05).结论 PPAR-γ的配体罗格列酮对原位肝移植胆道缺血-再灌注损伤有保护作用,这种保护作用的机制与拮抗核因子-κB和抑制其表达,减少下游IL-6,IL-1β以及TNF-α等细胞因子的释放有关.

abstractsObjective To explore the effective molecular mechanism of PPAR-γligands rosiglitazone to biliary ischemia-reperfusion injury in autologous liver transplantation. Method A total of 40 SD rats were randomly (random number) divided into sham operation group (SO), ischemia - reperfusion group (Ⅰ/R), rosiglitazone (ROS) and GW9662 group, with 10 ones in each. The models, rat biliary ischemiareperfusion injury of autologous liver transplantation, were made by modified two-cuff technique. Tissues of the liver and bile ducts and blood of those models were evaluated by pathological and biochemical methods to make sure the models were made successfully or not. SO group suffered autologous orthotopic liver transplantation, and L/R group suffered both that and ischemia-reperfusion. ROS group were injected rosiglitazone (0.3mg/kg) via portal vein after having been done all as I/R. GW9662 group suffered all as ROS, and 10min later ,they were injected GW9662(0.3mg/kg) via portal vein. 4h after the experiment, tissues of livers and bilary ducts were taken to be tested by immunohistochemistry method, and the blood punctured from the right ventricular were taken to be determined by ELISA. ANOVA was used for statistical analysis.Results IL-1β, TNF-α and IL-6 were mainly expressed in the cytoplasm of hepatocytes and bile duct cells,while NF-κB was expressed both in the cytoplasm and nuclei. Expression of those proteins in L/R and GW9662 group was increased, significantly higher when compared to the SO and ROS (P ＜ 0.05). IL-1β,TNF-α and IL-6 in rat serum were simultaneously increased, and significantly higher than SO(P ＜0.05).Compared with the SO, expressions of the IL-1 β,TNF-α and IL-6 were not significantly changed in ROS (P＞ 0.05 )but significantly increased in GW9662. Conclusions PPAR-γ ligand rosiglitazone took protective role in biliary ischemia-reperfusion injury in autologous liver transplantation. The mechanism correlates with the release of the IL-lα, IL-1β and TNF-α and other inflammatory mediators, which decreased as the expression of NF-κB inhibited by its antagonist.