脑死亡供肝中乙醛脱氢酶表达与细胞凋亡的关系

Relationship Between the Expression of ALDH2 and Apoptosis Level in DBD Liver

目的:探究脑死亡供肝中乙醛脱氢酶(ALDH2)表达与细胞凋亡的关系,为临床脑死亡供肝质量评估提供新指标。方法:应用免疫组化学方法检测脑死亡家兔肝脏组织中p-ALDH2表达;细胞水平建立缺氧模型并应用抑制剂大豆苷或激动剂Alda-1干扰ALDH2表达,应用免疫印迹法检测p-ALDH2和Cleaved-caspase 3的表达。结果:p-ALDH2主要表达于内皮细胞和肝细胞,且脑死亡组中p-ALDH2表达较低;人脐静脉内皮细胞中缺氧组pALDH2表达水平较正常组低而Cleaved-caspase 3较高;大豆苷+缺氧组中p-ALDH2表达降低,Cleaved-caspase 3表达升高;在Alda-1+缺氧组中,与正常组相比,p-ALDH2的表达水平较低,Cleaved-caspase 3表达水平较高,而与缺氧组相比,p-ALDH2的表达水平较高,Cleaved-caspase 3表达水平较低,差异均具有统计学意义(P<0.05)。结论:脑死亡供肝ALDH2表达降低可增加细胞凋亡,引起肝脏损伤。

Objective：To explore the relationship between the expression of ALDH2 and apoptosis level in donor after brain death（DBD）liver and provide a new indicator of evaluating DBD liver for clinic.Methods：Immunohistochemical methods were adopted to detect the expression of p-ALDH2.After the expression of ALDH2 in HUVECS during hypoxia had been inhibited or activated,the expression of p-ALDH2 and Cleaved-caspase 3 were analyzed using Western blot.Results：Immunohistochemistry showed that p-ALDH2 was primarily located in endothelial and hepatocyte,and the brain death group had a lower expression of p-ALDH2.Following the treatment of inhibitor of ALDH2,daidzin,in combination with hypoxia for 8hours,the p-ALDH2 decreased and cleaved-caspase 3significantly increased in contrast to that in hypoxic HUVECs,however,the p-ALDH2 increased and cleaved-caspase 3decreased after treatment with Alda-1and hypoxia compared with that in hypoxic HUVECs（P〈0.05）.Conclusion：Decreased ALDH2 led to apoptosis in DBD liver,and led to injury of liver.