肺泡巨噬细胞Toll样受体4在肝脏缺血再灌注损伤小鼠肺组织中的表达及意义  

Expression and significance of alveolar macrophage Toll-like receptor 4 in pulmonary tissues during total hepatic ischemia/reperfusion injury in mice

作　　者：徐建波[1] 吴河水[1] 张进祥[2] 王琳[3] 杨鹏[1] 田元[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院普外科,武汉430022 [2]华中科技大学同济医学院附属协和医院急诊外科,武汉430022 [3]华中科技大学同济医学院附属协和医院儿科,武汉430022

出　　处：《中华肝胆外科杂志》2008年第2期115-117,共3页Chinese Journal of Hepatobiliary Surgery

基　　金：国家自然科学基金资助项目（30200272）

摘　　要：目的探讨肺泡巨噬细胞Toll样受体4在肝脏缺血再灌注损伤小鼠肺组织中的表达及意义。方法以野生型小鼠C3h／Heouj及TLR4缺失小鼠C3h／Hej复制全肝脏缺血再灌注损伤模型，分别于缺血20min再灌注1h，6h时经支气管肺泡灌洗（bronchoalveolar lavage fluid，BAIJ）获取肺泡巨噬细胞和肺泡灌洗液，采用免疫组织化学法检测肺泡巨噬细胞表面TLR4的表达，并以鲎试剂内毒素检测试剂盒和酶联免疫吸附试验（ELISA）检测灌洗液中内毒素及肿瘤坏死因子α（TNF-α）的水平。同时检测肺组织湿／干重比值、肺组织髓过氧化物酶（myeloperosidase，MPO）的含量以及计算肺组织学评分。结果（1）与假手术组小鼠相比，野生型小鼠（C3h／Heouj）在缺血再灌后各时间点肺泡巨噬细胞Toll样受体4蛋白表达升高，6h强于1h，且支气管肺泡灌洗液中TNF-α的水平明显升高，肺组织湿／干重比值持续升高，MPO的含量持续增加（P〈0.05）。（2）与C3h／Heouj小鼠相比，C3h／Hej组小鼠在再灌注后支气管肺泡灌洗液中TNF-α的含量明显降低（P〈0．05），肺损伤明显减轻（P〈0.05），肺组织湿／干重比值及MPO的含量明显下降（P〈0.05）。（3）各组小鼠缺血再灌注后1h肺泡灌洗液中内毒素水平与假手术组相比，差异无统计学意义（P〉0.05），而在6h时则明显升高，差异有统计学意义（P〈0.05）。结论小鼠全肝缺血再灌注过程中，肺泡巨噬细胞表面Toll样受体4激活，并与肺功能损伤有关。Objective To explore the activation and its significance of Toll-like receptor 4 on alveolar macrophage of the lungs during the process of total hepatic ischemia/reperfusion injury in mice. Methods Wild type mice（C3h/Heouj）and TLR4-defieient mice（C3h/HeJ） were used in a model of total hepatic ischemia/reperfusion. Alveolar macrophages were collected at the time phases of 1 h and 6 h by the means of bronchoalveolar lavage （BAL） after 20-minute block of total blood flow of the liver. The expression of TLR4 was observed with immunohistochemical staining. The levels of endotoxin and TNF-α in BAL fluid were determined. The concentration of MPO, ratio of Wet/dry, and lung histological scores were measured. Results Compared with TLR4 deficient （C3h/Hej） animals, the expression of TLR4 in I/R group were up-regulated in wild type mice （C3h/Hej） at the time point of 1st and 6th h of reperfusion after 20 minutes of total liver ischemia, and markedly increased at the 6th h. Meanwhile, the concentrations of TNF a and MPO, the ratio of Wet/dry and lung histological scores were increased. TNF a level was significantly decreased as compared to Wild type mice at both time points after reperfusion （P〈0.01）, and lung injury was found to be highly reduced（P〈0.05）. In two strains of animals, the endotoxin level in BALF was increased a little without no significances in I/R group as compared with Sham group 1 h after the reperfusion （P〉0. 05）, and markedly increased at the 6th hours （P〈0.05）. Conclusion Toll like receptor 4 on alveolar macrophage is activated in the process of total hepatic ischemia/reperfusion and involved in the pulmonary injury.

关 键 词：再灌注损伤 TOLL样受体 肝脏 肺损伤 肺泡巨噬细胞 

分 类 号：R686[医药卫生—骨科学]