不同粒径汽车尾气颗粒物对A549细胞TNF-α等炎性因子及NF-κB的影响  

Effects of automobile exhaust particulates with different sizes on TNF-α and other inflammatory factors and NF-κB in A549 cells

作　　者：徐雯[1] 任飞林[1] 黄春妍[1] 戴兵[1] 梅德清[2] 江俊康[1] 

机构地区：[1]南通大学公共卫生学院,江苏南通226019 [2]江苏大学汽车与交通工程学院

出　　处：《环境与健康杂志》2012年第11期1013-1016,共4页Journal of Environment and Health

基　　金：国家自然科学基金(50976051);交通运输部联合科技攻关项目(2009-353-332-280);江苏省交通运输厅科技创新攻关计划项目(10Y25)

摘　　要：目的探讨并比较不同粒径汽车尾气颗粒物对人肺癌上皮细胞A549肿瘤坏死因子-α（TNF-α）、白介素-6（IL-6）、核转录因子-κB（NF-κB）的mRNA及NF-κB蛋白表达的影响。方法以不同粒径（0．18-3．2μm）汽车尾气颗粒物混悬液（100μg／m1）染毒A549细胞，48h后RT-PCR法检测细胞TNF-α、IL-6及，NF-κB（用5）基因表达水平，Western Blot法检测胞浆及胞核内NF-κB（P65）蛋白表达水平。结果与对照组相比，各染毒组细胞的TNF-仪、1L-6及NF-κB（P65）mRNA表达水平均上升（P〈0．05），由高到低依次为0．56～、0．32～、0．18～、1．0～和1．8～3．2μm组，两两之间（除0．18～和0．32～μm组之间、1．0～和1．8～3．2μm组之间）差异均有统计学意义（P〈O．05）；各组TNF-OL、IL-6mRNA水平与NF-κB（P65）mRNA水平显著相关（r=0．944，0．890，P〈0．05）。与对照组相比，除1．8～3．2μm组无统计学意义外，其余各组胞浆NF-κB（P65）蛋白水平均降低（P〈0．05），由高到低依次为1．0～、0．18～、0．32～、0．56～μm组；而胞核NF-κB（P65）蛋白水平均升高（P〈0．05），由低到高依次为1．0～、0．18～、0．32～、0．56～μm组。结论不同粒径汽车尾气颗粒物作用于A549细胞后均可使TNF-α、IL-6、NF-κBmRNA表达升高、NF-κB活化，活化的NF-κB由细胞质进入细胞核发挥其调控炎症反应的作用。各染毒组中以0．56～μm组炎性损伤效应最强，粒径是影响颗粒物毒性作用大小的重要因素之一。Objective To investigate the effects of automobile exhaust particulates with different sizes on TNF-α,IL-6, NF-κB mRNA expression and the protein expression of NF-κB in human lung adenocarcinoma cells A549. Methods A549 cells were treated with automobile exhaust particulates collected with a particle size classifier at 100μg/ml for 48h. RT-PCR method was used to detect the changes of mRNA expression in TNF-α,IL-6 and NF-κB （P65） genes. Western Blot analysis was performed to detect the expression of NF-κB （P65） both in cytoplasm and nucleus. Results A549 cells exposed to automobile exhaust particulates showed significant increases in TNF-α,IL-6 and NF-κB （P65） mRNA expression compared with the control（P〈0.05）, and the most conspicuous effect was found in the 0.56-μm group （over one-fold）, then followed by 0.18-, 0.32-,1.0- and 1.8-μ.m groups, respectively. There were statistical siguificances in the pairwise comparison between each two exposure groups （P〈O.05）, except 0.18- and 0.32-, 1.0- and 1.8-μ.m groups （P〉0.05）. The levels of TNF-α, IL-6 mRNA correlated significantly with corresponding NF-κB （P65） mRNA expression （r=0.944, 0.890, P〈0.05）. NF-κB （P65） protein expression in the cytoplasm after exposure was found to be significantly reduced（P〈0.05）, except 1.8-3.2 μ.m group, the descending order was 1.0-, 0.32-, 0.18- and 0.56-μm groups. By contrast, NF-κB （P65） protein expression in the nucleus was all significantly increased, and the order was 1.0-,0.32-,0.18- and 0.56-μm groups. Conclusion A549 cells can be stimulated by automobile exhaust particles with different sizes and make TNF-α,IL-6 and NF-κB mRNA expression up- regulated and lead activated NF-κB getting into the cell nucleus from cytoplasm. There is the strongest inflammatory injury in 0.56-μm group and the particulate size may be one of the factors influencing the adverse effects of particulates on human health.

关 键 词：汽车尾气颗粒物 粒径 A549细胞 肿瘤坏死因子-Α 白介素-6 核转录因子-ΚB 基因表达 蛋白表达 

分 类 号：R122.2[医药卫生—环境卫生学]