抗TNF-α单克隆抗体对大鼠肝肾联合移植术后肝肾细胞凋亡的影响

Role of Tumor Necrosis Factor-Alpha Monoclonal Antibody in Cell Apoptosis after Combined Liver and Kidney Transplantation in Rats

目的探讨大鼠肝肾联合移植术后抗TNF-α单克隆抗体对移植肝肾组织细胞凋亡的影响。方法建立大鼠肝肾联合移植模型,80只大鼠按完全随机法均分为受体组与供体组,两两随机配对后均分为实验组和对照组。实验组:灌注4℃的生理盐水40ml(其中供肝经门静脉冷灌注30ml,供肾经肾动脉冷灌注10ml)+抗TNF-α单克隆抗体(0.1mg/kg体重);对照组:灌注等量4℃的生理盐水。于灌注后不同时点取静脉血检测肝肾功能,采用酶联免疫吸附(ELISA)法和原位DNA片断末端标记(TUNEL)法,检测移植肝肾组织中TNFα的含量及细胞凋亡情况。结果实验组血AST、ALT、Cr和BNN含量及移植肝肾组织中TNFα水平均明显低于对照组相应时相(P<0.05);实验组移植肝肾组织中细胞凋亡指数明显低于对照组相应时相(P<0.05);实验组移植肝肾组织病理改变不明显,组织结构基本正常。结论在肝肾联合移植过程中,应用抗TNFα单克隆抗体可减少移植肝肾组织内细胞凋亡,促进移植肝肾功能恢复。

Objective To investigate the potential role of tumor necrosis factor-alpha （TNF-α） in apoptosis after combined liver and kidney transplantation in rats. Methods Eighty rats which had combined liver and kidney transplantation were randomly paired, were divided into study group （n 20） and control group （n=20）. 40 ml of 4℃ sodium chloride and anti-TNF-α monoclonal antibody （30 ml was infused from portal veins to donated livers and 10 ml from renal arteries to donated kidneys） were infused to the study group （0. 1 mg/kg weight）,and the same quantity of 4℃ sodium chloride was infused the control group. Venous blood was drew at different phases after the transplantations to detect the function of kidney and liver. The level of TNF-α and the cell apoptosis were detected in the transplanted tissues of liver and kidney by ELISA and terminal deoxynucleotidy transferase mediated dTUP-biotin nick-end labeling （TUNEL）. Results The levels of AST, ACT, Cr and BUN in the study group were significantly lower than those of the control group at the same phases （P〈0.05）. The level of TNF-α in the transplanted tissues of kidney and liver was also significantly lower as compared with those of control group. The cell apoptosis index of the transplanted tissues of kidney and liver was significantly smaller in the study group （P〈0.05）. There was no dramatically pathological change in the tissues of transplanted kidney and liver, which were treated with anti- TNF-α monoclonal antibody, and the structures are almost normal. Conclusion Anti-TNF-α monoclonal antibody may reduce cell apoptosis and accelerate the restoration of function of liver and kidney after combined liver and kid- ney transplantation.