【摘要】 目的研究采用肿效瘤坏死因子-α(TNF-α)预处理与采用缺血预处理两种方法对减轻大鼠肝脏缺血再灌注损伤(IRI)的作用效果,并探讨采用TNF-α预处理减轻大鼠肝脏IRI的机制。方法将40只Wistar大鼠随机分为4组:假手术组(SO组)仅行开腹及游离肝十二指肠韧带;缺血再灌注组(IR组)采用Pringle’s法阻断肝门30 min,再灌注6 h;缺血预处理组(IPC组)采用Pringle’s法阻断肝门10 min,开放血流10 min,此后操作同IR组;TNF-α预处理组(TPC组),术前30 min给予TNF-α1μg/kg腹腔注射,此后操作同IR组。检测血清谷丙转氨酶(ALT)和谷草转氨酶(AST),采用免疫组织化学检测Bcl-2蛋白和NF-κB p65蛋白表达情况,以及肝细胞凋亡指数(AI)。结果 IPC组血清ALT、AST水平为(316.4±90.6)U/L、(316.4±90.6)U/L,TPC组为(336.8±79.4)U/L、(392.7±109.3)U/L,与IR组(642.8±149.3)U/L、(730.6±103.0)U/L比较明显降低,P<0.05差异有统计学意义;肝细胞凋亡评分AI在IPC组(4.36+0.88)和TPC组(4.94+2.04)明显降低,与IR组(9.48+2.42)比较,P<0.05差异有统计学意义;肝组织内Bcl-2蛋白阳性表达在IPC组(62.30+9.20)和TPC组(60.99+6.30)升高,与IR组(11.45+11.97)比较,有显著性差异(P<0.05);NF-κB p65阳性表达在IPC组(31.56+4.85)和TPC组(32.80+5.30)明显降低,与IR组(68.33+6.07)比较,有显著性差异(P<0.05)。上述各项指标在IPC组与TPC组间,差异无统计学意义(P>0.05)。结论采用TNF-α预处理与缺血预处理对减轻大鼠肝脏IRI的效果一致,TNF-α预处理通过抑制NF-κB p65蛋白表达、激活凋亡抑制基因Bcl-2蛋白表达,减轻了大鼠肝脏IRI。更多还原

【Abstract】 Objective To compare the effect of TNF-α preconditioning and ischemic preconditioning on hepatic ischemia-reperfusion injury( IRI) and investigate the underlying mechanisms of TNF-α preconditioning. Methods Fourty healthy male Wistar rats were randomly divided into four groups which were Sham-operated group( SO),ischemia-reperfusion group( IR group: produced by total inflow occlusion for 30 min),ischemic preconditioning group( IPC group: induce with 10 min hepatic ischemic and open 10 min before IR) and TNF-α preconditioning group( TPC group: intraperitoneal injection with 1 μg / kg TNF-a 30 min prior to IR). The sample of blood and hepatic tissue of all groups were taken after experiment. The protein levels of NF-κBp65 and Bcl-2 in the hepatic tissue were detected by immunohistochemistry. Results There was significant difference( P < 0. 05) between IR group and IPC group,TPC group on the level of ALT,AST and the expression of NF-κBp65 and Bcl-2,apoptosis index in hepatic tissue. There was no significance difference( P > 0. 05) between IPC group and TPC group. Conclusion TNF-α preconditioning decreased the intensity of hepatic IRI,just as ischemic preconditioning,by induces an decrease in the NF-κBp65 expression and an increase in the Bcl-2 expression.更多还原