中华急诊医学杂志
中華急診醫學雜誌
중화급진의학잡지
CHINESE JOURNAL OF EMERGENCY MEDICINE
2011年
2期
163-167
,共5页
裴红红%张正良%柏玲%白郑海%缪菲
裴紅紅%張正良%柏玲%白鄭海%繆菲
배홍홍%장정량%백령%백정해%무비
过氧化物酶体增殖体-γ%罗格列酮%原位肝移植%胆道损伤%缺血-再灌注%移植肝缺血型胆道病变%核因子-κB%全身炎症反应综合征
過氧化物酶體增殖體-γ%囉格列酮%原位肝移植%膽道損傷%缺血-再灌註%移植肝缺血型膽道病變%覈因子-κB%全身炎癥反應綜閤徵
과양화물매체증식체-γ%라격렬동%원위간이식%담도손상%결혈-재관주%이식간결혈형담도병변%핵인자-κB%전신염증반응종합정
PPAR-γ%Rosiglitazone%Aotologous liver transplantation%Biliary injury%Ischemiareperfusion%lschemic type biliary lesion%NF-κB%Systemic inflammatory response syndrome
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.
目的 探討過氧化物酶體增殖物活化受體-γ(PPAR-γ)配體囉格列酮在原位肝移植膽道缺血-再灌註損傷中作用的分子機製.方法 40隻SD大鼠隨機(隨機數字法)分成假手術組(SO組)、缺血-再灌註組(I/R組)、囉格列酮組(ROS組)和GW9662組,每組10隻.通過改良"雙袖套法"建立大鼠自體原位肝移植膽道缺血-再灌註模型,通過肝髒及膽管組織病理學變化、血生化指標檢測評價模型建立是否成功.SO組行自體原位肝移植術,I/R行建模缺血-再灌註,ROS在缺血-再灌註後以囉格列酮0.3mg/kg經門靜脈註射,GW9662在ROS基礎上10 min後經門靜脈以0.3mg/kg註射GW9662,各組均于實驗後4h取部分肝髒和膽管組織用于免疫組化檢測;右心室穿刺抽血採用ELISA法測定細胞因子含量.採用方差分析進行統計學處理.結果 組織中細胞因子IL-1β,TNF-α,IL-6活性主要錶達在肝細胞及膽管細胞胞漿中,轉錄因子NF-κB則在胞漿及胞覈中均有錶達;I/R組及GW9662組上述蛋白錶達升高,和SO組以及ROS組比較明顯增高(P＜0.05).血清中I/R組大鼠IL-1β,TNF-α及IL-6同步升高,較SO組明顯增高(P＜0.05);囉格列酮榦預後血清中IL-1β,TNF-α及IL-6和SO組比較無明顯變化(P＞0.05);給予囉格列酮阻滯劑GW9662後,IL-1β,TNF-α及IL-6的含量和SO組比較則明顯增高(P＜0.05).結論 PPAR-γ的配體囉格列酮對原位肝移植膽道缺血-再灌註損傷有保護作用,這種保護作用的機製與拮抗覈因子-κB和抑製其錶達,減少下遊IL-6,IL-1β以及TNF-α等細胞因子的釋放有關.
목적 탐토과양화물매체증식물활화수체-γ(PPAR-γ)배체라격렬동재원위간이식담도결혈-재관주손상중작용적분자궤제.방법 40지SD대서수궤(수궤수자법)분성가수술조(SO조)、결혈-재관주조(I/R조)、라격렬동조(ROS조)화GW9662조,매조10지.통과개량"쌍수투법"건립대서자체원위간이식담도결혈-재관주모형,통과간장급담관조직병이학변화、혈생화지표검측평개모형건립시부성공.SO조행자체원위간이식술,I/R행건모결혈-재관주,ROS재결혈-재관주후이라격렬동0.3mg/kg경문정맥주사,GW9662재ROS기출상10 min후경문정맥이0.3mg/kg주사GW9662,각조균우실험후4h취부분간장화담관조직용우면역조화검측;우심실천자추혈채용ELISA법측정세포인자함량.채용방차분석진행통계학처리.결과 조직중세포인자IL-1β,TNF-α,IL-6활성주요표체재간세포급담관세포포장중,전록인자NF-κB칙재포장급포핵중균유표체;I/R조급GW9662조상술단백표체승고,화SO조이급ROS조비교명현증고(P＜0.05).혈청중I/R조대서IL-1β,TNF-α급IL-6동보승고,교SO조명현증고(P＜0.05);라격렬동간예후혈청중IL-1β,TNF-α급IL-6화SO조비교무명현변화(P＞0.05);급여라격렬동조체제GW9662후,IL-1β,TNF-α급IL-6적함량화SO조비교칙명현증고(P＜0.05).결론 PPAR-γ적배체라격렬동대원위간이식담도결혈-재관주손상유보호작용,저충보호작용적궤제여길항핵인자-κB화억제기표체,감소하유IL-6,IL-1β이급TNF-α등세포인자적석방유관.
Objective 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.
