缺血后处理对大鼠肝缺血再灌注时肝细胞线粒体膜通透性转换和膜电位的影响  

Effects of ischemic postconditioning on mitochondrial permeability transition and mitochondrial transmembrane potential following hepatic ischemia-reperfusion in rats

作　　者：朱宇麟[1] 李静[2] 吕毅[3] 景桂霞[1] 

机构地区：[1]西安交通大学医学院第一附属医院麻醉科,710061 [2]西安市红十字会医院麻醉科 [3]西安交通大学医学院第一附属医院肝胆外科

出　　处：《中华麻醉学杂志》2011年第3期364-367,共4页Chinese Journal of Anesthesiology

基　　金：陕西省科技攻关项目（2008K14-02）

摘　　要：目的评价缺血后处理对大鼠肝缺血再灌注时肝细胞线粒体膜通透性转换和膜电位（△ψm）的影响。方法成年健康雄性SD大鼠40只，体重220—260g，采用随机数字表法，将其随机分为5组（n=8）：假手术组（S组）、苍术苷＋假手术组（A＋S组）、缺血再灌注组（IR组）、缺血后处理组（IPO组）和苍术苷＋缺血后处理组（A＋IPO组）。采用阻断肝中叶和左叶60min，恢复血流灌注6h的方法建立大鼠肝缺血再灌注模型。s组和A＋s组仅游离肝门，不阻断血管；A＋s组关腹前静脉注射苍术苷5mg／kg；IR组制备肝缺血再灌注模型；IPO组于再灌注前行缺血后处理，再灌注1min，缺血1min，反复3次；A＋IPO组于再灌注前静脉注射苍术苷5mg／kg。于缺血前即刻和再灌注6h时，采集左颈静脉血样，测定血清ALT和AST的活性。再灌注6h时处死大鼠，取肝左叶组织，观察超微结构和细胞凋亡情况，计算凋亡指数，测定细胞色素c（Cyte）的表达水平、△ψm和线粒体通透性转换孔（MPrP）活性。结果与s组比较，A＋S组时血清ALT和AST的活性、凋亡指数、Cytc表达、△ψm和MPTP活性差异无统计学意义（P〉0．05），IR组、IPO组和A＋IPO组再灌注6h时血清ALT和AST的活性、凋亡指数升高，Cytc表达上调，△ψm降低，MPTP活性升高（P〈0．05）；与IR组比较，IPO组血清ALT和AST的活性、凋亡指数降低，Cytc表达下调，△ψm升高，MPTP活性降低（P〈0．05），肝组织病理学损伤减轻，A＋IPO组各指标差异无统计学意义（P〉0．05）；与IPO组比较，A＋IPO组血清ALT和AST的活性、凋亡指数升高，Cytc表达上调，△ψm降低，MPTP活性升高（P〈0．05），肝组织病理学损伤加重。结论缺血后处理可抑制肝细胞线粒体膜通透性转换，减少线粒体△ψm的耗散，从而减轻大鼠肝缺血再灌注损伤。Objective To investigate the effects of ischemic postconditioning on mitochondfial permeability transition and mitochondfial transmembrane potential （△ψm） following hepatic ischemia-reperfusion （I/R） in rats. Methods Forty male SD rats weighing 220-260 g were randomly divided into 5 groups with 8 animals in each group ： sham operation group （group S） ; atractyloside ＋ sham operation group （group A ＋ S） ; I/R group; ischemic postconditioning group （group IPO） and atractyloside ＋ ischemic postconditioning group （group A ＋ IPO）. The ani- reals were anesthetized with intramuscular injection of atropine 0.05 mg/kg. Hepatic I/R was produced by occlusion of hepatic blood flow for 60 min followed by 6 h reperfusion. In group A ＋ S, atractyloside 5 mg/kg was injected intravenously before abdomen was closed. In group IPO, the animals were subjected to 3 cycles of 1 min reperfusion interspersed with 1 min hepatic ischemia at the end of 60 min hepatic ischemia. In group A ＋ IPO, atractylo-side 5 mg/kg was injected intravenously before reperfusion. Venous blood samples were collected for determination of serum ALT and AST activities immediately before ischemia and at 6 h of reperfusion. The animals were then sacrificed. Their livers were removed for microscopic examination, detection of apoptosis and determination of cyto- chrome c （Cyt c） expression,△ψm and mitochonerial permeability transition pore （MPTP） activity. Apeptosis index （AI） was calculated.Results There was no significant difference in serum ALT and AST activities, AI, Cyt c expression, △ψm and MPTP activity between S and A ＋ S groups （ P 〉 0.05 ）. Compared with group S, serum ALT and AST activities and AI were significantly increased, Cyt c expression was up-regulated, △ψm was de- creased and MPTP activity was increased in groups I/R, IPO and A ＋ IPO （ P 〈 0.05）. Compared with group I/R, serum ALT and AST activities and AI were significantly decreased, Cyt c expression was down-regulat

关 键 词：再灌注损伤 线粒体 肝 细胞膜通透性 膜电位 缺血后处理 

分 类 号：R285.5[医药卫生—中药学]