活性氧对尘肺患者肺泡巨噬细胞凋亡信号转导途径的作用

Effect of reactive oxygen species on the alveolar macrophages apoptosis of pneumoconiosis cases

目的探讨活性氧(ROS)对Fas/FasL信号转导途径致肺泡巨噬细胞凋亡作用。方法以中国北戴河煤炭工人疗养院无其他肺部疾病的Ⅰ期、Ⅱ期尘肺患者共45例为研究对象,将支气管肺泡灌洗液(BALF)中的肺泡巨噬细胞(AM)分两组培养,即对照组、超氧化物歧化酶(SOD)组。采用TUNEL技术检测AM凋亡情况,琼脂糖凝胶电泳法检测DNA片段,W estern b lot法检测AM的Fas、FasL、caspase-8、caspase-3表达水平。结果SOD组的凋亡指数(9.50±2.76)显著低于对照组(19.18±2.83,P<0.05);SOD组的Fas、FasL、caspase-8、caspase-3蛋白表达水平(分别为0.107±0.065、0.115±0.047、0.231±0.139、0.312±0.114)均低于对照组(分别为0.209±0.084、0.210±0.097、0.374±0.071、0.441±0.095,P<0.05);对照组出现凋亡细胞特征性"梯状带",而SOD组未出现。结论Fas/FasL信号转导途径介导的AM凋亡可能与ROS上调Fas/FasL系统的表达有关。

Objective To explore the effect of reactive oxygen species （ROS） on alveolar macrophages apoptosis induced by the Fas/FasL signal transduction. Methods Phase Ⅰ and Ⅱ pneumoconiosis cases without the other lung diseases in Beidaihe Coal Workers＇ Sanatorium were as the study subjects. Macrophages（AM） of BALF were cultivated in 2 groups： control group and SOD group. The apoptosis indexes of alveolar AM were detected by TUNEL method, DNA fragment was detected by agarose gel electrophoresis（AGE）, and the expressions of Fas, FasL, caspase-8, caspase-3 were detected by western blotting. Results The apoptosis indexes of SOD group （ 9. 50 ± 2. 76 ） was much lower than those of the control group （ 19. 18 ± 2. 83 ） （ P 〈 0.05）. The expressions of Fas, FasL, caspase-8, caspase-3 of SOD group （0. 107±0.065, 0. 115±0.047, 0.231±0. 139 and 0. 312±0. 114） were much lower than those of the control group （0. 209±0. 084, 0. 210±0. 097, 0. 374±0. 071 and 0. 441±0. 095 ,P 〈0. 05 ）. DNA ladder emerged in the control group but did not appear in SOD group. Conclusion The alveolar macrophages apoptosis induced by Fas/FasL signal transduction could be associated with the expression enhancement of Fas induced by ROS.