缺血预处理对大鼠减体积肝移植缺血再灌注损伤的影响

Effects of ischemia precondition on ischemia reperfusion injury of graft after reduced-size liver transplantation of rats

目的观察缺血预处理(ischemia preconditioning,IPC)对大鼠减体积肝移植后肝组织氧化还原因子-1(Redox factor-1,Ref-1)蛋白表达及术后肝脏损伤的影响。方法将100只Lewis成年雄性大鼠随机分成3组:缺血预处理组(IPC组)、50%移植组(PLT组)和假手术组(SO组),分别于移植后0.5、2、6和24h取材,通过Western免疫杂交法和免疫组织化学技术结合图像分析定量检测移植后各时间点Ref-1蛋白表达变化,同时结合血清学和组织病理学分析Ref-1表达变化的意义。结果IPC可使减体积肝移植后早期肝实质细胞中Ref-1蛋白表达增高。与PLT组相比,IPC组早期Ref-1蛋白表达明显升高(P<0.05)。病理学分析显示PLT组术后24h可见到门脉周围大量炎细胞浸润,肝窦扩张明显,肝组织损伤较重;而IPC组则损伤较轻。术后6和24h血清ALT值分别为:PLT组(1186.65±142.31)u/L;(1498.91±126.79)u/L;IPC组(799.61±125.97)u/L;(659.27±135.68)u/L。与PLT组相比,IPC组术后6和24h血清ALT值明显降低(P<0.05;P<0.01)。结论IPC可减轻减体积肝移植后早期肝脏的损伤,其机制可能与促进Ref-1蛋白的表达有关。

[Objective] To observe the effects of ischemia precondition on ischemia reperfusion injury of graft after reduced-size liver transplantation of rats as well as on the expression of Ref-1 protein in order to explore the protection mechanisms of IPC against liver injury. [Methods] 120 adult male Lewis rats were randomly divided into three groups： 50% partial liver transplantation （PLT group）, 50% partial liver transplantation with ischemia preconditioning （IPC group） and sham operation group （SO group）. Animals were sacrificed in each group at different time points： 0.5, 2, 6 and 24 h after liver transplantation. Then, we explored the changes and significance of the expression of Ref-1 according to Western blotting and immunohistochemistry, associated with serology and histopathology analysis. [Results] As compared to SO group, our results revealed the increased expression of Ref-1 protein in both IPC group and PLT group. The expression of Ref-1 protein in IPC group was higher than that in PLT group in earlier period after liver transplantation （P 〈0.05）. With pathobiological analysis, we found there were lots of inflammation cells infiltration around the portal veins, the serious sinus hepaticus dilation and damage of hepatic tissue. However, the damage in IPC group were comparatively slight. The serum ALT values of animal at 6h and 24 h after liver transplantation in PLT group and IPC group were （1186.65±142.31） u/L; （1498.91±126.79） u/Land （799.61±125.97）U/L; （659.27±135.68）U/L. As compared to PLT group, the ALT values at 6h and 24h after operation in IPC group decreased significantly （P 〈0.05; P 〈0.01）. [Conclusions] These data suggests that ischemia preconditioning relieves the liver injury after reduced-size liver transplantation in earlier period, which is partly at least relevant with the promotion of Ref-1 protein expression.