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目的 阐明羟自由基 ( OH)、细胞凋亡及Bcl 2蛋白表达在大鼠肝脏不同保存时相再灌注过程中的变化规律及相互关系。方法 以大鼠肝脏离体非循环灌注为模型 ,用高效液相色谱(HPLC)紫外检测法 ,测定 OH与水杨酸的生成物二羟苯甲酸 (DHBA)含量 ,用原位末端标记法及电镜观察细胞凋亡 ,用免疫组织化学法检测Bcl 2蛋白表达。结果 随着保存时间延长 (18h内 ) , OH水平及细胞凋亡指数 (AI)进行性升高 ,且两者呈显著正相关 ;而Bcl 2蛋白表达指数 (BI)则进行性降低 ,BI与 OH水平及AI呈显著负相关。结论 OH及细胞凋亡同时参与肝脏保存再灌注损伤 ,Bcl 2蛋白对细胞凋亡具有抑制作用。
OBJECTIVE: To elucidate the regularity and relationship between the changes of hydroxyl free radicals (OH), apoptosis and the expression of Bcl 2 protein during reperfusion of different preservation phases in rat liver. Methods The isolated rat liver acyclic perfusion model was used to determine the content of dihydroxybenzoic acid (DHBA) of OH and salicylic acid by high performance liquid chromatography (HPLC) Cell apoptosis was observed by electron microscope, and the expression of Bcl2 protein was detected by immunohistochemistry. Results With the prolongation of storage time (within 18 hours), the level of OH and the apoptosis index (AI) increased progressively, and there was a significant positive correlation between the two. However, the Bcl 2 protein expression index (BI) OH level and AI was significantly negatively correlated. Conclusion OH and apoptosis are both involved in hepatic preservation and reperfusion injury, and Bcl 2 protein has an inhibitory effect on apoptosis.