大鼠肝移植急性排斥反应中肿瘤坏死因子受体配体的表达及其意义  

Expression of glucocorticoid induced tumor necrosis factor receptor ligand during acute rejection after rat liver transplantation

作　　者：黄中荣[1] 魏思东[2] 李金政[2] 刘作金[2] 龚建平[2] 余正[1] 

机构地区：[1]重庆医科大学附属永川医院外科,402160 [2]重庆医科大学附属第二医院肝胆外科重庆市肝胆外科重点实验室

出　　处：《中华实验外科杂志》2011年第8期1321-1323,共3页Chinese Journal of Experimental Surgery

基　　金：基金项目：国家自然科学基金资助项目（30772098、30972888、81070374、30801126）;重庆市卫生局资助项目（2010-2-260）

摘　　要：目的观察肝移植排斥反应中移植肝脏内糖皮质激素诱导的肿瘤坏死因子受体配体（GITRL）的表达。方法采用Kamada’s二袖套法建立从Lewis到Brown Norway（BN）大鼠的肝移植排斥模型为排斥组（n=5），从BN到BN的肝移植模型为耐受组（n=5）。术后24h，抽取血液，取肝脏及分离库普弗（Kupffer）细胞，检测肝脏上GITRL及肿瘤坏死因子（TNF）-α的表达，Kupffer细胞上GITRL的表达，检测血清及细胞上清液中TNF-仪的表达。免疫组织化学染色强度采用Image-Pro Plus6．0图像分析软件分析。结果免疫耐受组和排斥组肝脏内的GITRL的平均染色强度分别为0．113±0．007和0．270±0．018（P〈0．05），TNF-α平均染色强度分别为0．114±0．004和0．141±0．005（P〈0．05），耐受组和排斥组的Kupffer细胞GITRL平均染色强度分别为0．206±0．017和0．337±0．018（P〈0．05），Kupffer细胞的培养上清液中，耐受组和排斥组TNF-α的值分别为（68．66-α21．12）、（178．33±29．39）ng／L（P〈0．05）。结论在排斥的早期阶段肝脏及Kupffer细胞的GITRL表达增高，监测和干扰GITRL可能有益于肝移植急性排斥反应的早期诊断和处理。Objective To investigate the changes of glucocorticoid induced tumor necrosis factor receptor ligand （GITRL） in hepatic allograft rejection. Methods Liver transplantation from Lewis rats （n = 5 ） to Brown Norway （BN） rats was performed by Kamada＇ s two-cuff technique as acute rejection group. Liver transplantation from BN to BN rats （ n = 5 ） was performed as tolerance group. Recipients were sacrificed at 24th h postoperation. Blood samples were collected and grafts were harvested, then Kupffer cells were isolated. GITRL and tumor necrosis factor （TNF）-α protein expression in the liver was tested by immunohistochemistry, and the GITRL expression in Kupffer cells by immunocytochemistry. Enzyme linked immunosorbent assay （ELISA） was employed to detect the changes of TNF-α protein in the serum and supernatant. The staining intensity was analyzed by Image-Pro Plus 6. 0 image analysis software. Results At 24th h postoperation, the liver GITRL expression levels in tolerance and rejection groups were 0. 113 ± 0. 007 and 0. 270 ± 0. 018, respectively （P 〈 0. 05 ）. The TNF-α expression levels in the liver in tolerance and rejection groups were 0. 114 ±0. 004 and 0. 141 ±0. 005 respectively （P 〈0. 05）. The GITRL expression levels in Kupffer cells in tolerance and rejection groups were 0. 206 ± 0. 017 and 0. 337±0 .018 respectively （P 〈0. 05）. As compared with tolerance group （68. 66 ±21.12） ng/L, TNF-α protein expression levels were up-regulated in the supernatant of rejection group （ 178. 33± 29. 39 ） ng/L （ P 〈 0. 05 ）. Conclusion The expression of GITRL in the liver and Kupffer cells was increased in the early stage of rejection, and monitoring and interfering GITRL may be useful for the early diagnosis and management of an acute rejection in liver transplantation.

关 键 词：肿瘤坏死因子受体配体 肝移植 急性排斥反应 KUPFFER细胞 

分 类 号：R657.3[医药卫生—外科学]