人肺癌A549细胞气-液界面暴露汽油发动机尾气中NO2的基准剂量评估  

Benchmark dose estimation of human lung cancer A549 cells exposed to NO_2 in the gasoline engine exhaust by air-liquid interface

作　　者：鱼涛[1] 宾萍[1] 钱旭东[2] 刘黎[1] 肖经纬[1] 李斌[1] YU Tao;BIN Ping;QIAN Xu-dong;LIU Li;XIAO Jmg-wei;LI Bin(Key Laboratory of Chemical Safety and Health, National Institute of Occupational Health and Poison Control, Chinese Center for Disease Control and Prevention, Beijing 100050, China;Department of Occupational and Radiation Hygiene, Changping Center for Disease Contrl and Prevention, Beijing 102200, China)

机构地区：[1]中国疾病预防控制中心职业卫生与中毒控制所,化学污染与健康安全重点实验室,北京100050 [2]北京市昌平区疾病预防控制中心职业与放射卫生科,北京102200

出　　处：《毒理学杂志》2018年第2期107-111,共5页Journal of Toxicology

基　　金：国家自然科学基金(81472955);中国疾病预防控制中心职业卫生与中毒控制所青年基金(2013DL-4)

摘　　要：目的通过气-液界面技术将人肺癌A549细胞暴露于不同工况下汽油发动机尾气,探讨汽油发动机尾气中的NO_2浓度与A549细胞存活率的剂量-效应关系及其基准剂量,为汽车尾气中NO_2的毒性效应风险评估提供基础数据。方法用气袋分别收集在怠速(冷启动)、53%和100%负荷下产生的汽油发动机尾气。采用盐酸萘乙二胺分光光度法检测汽油发动机尾气中NO_2的浓度;应用气-液界面技术将人肺癌A549细胞暴露于汽油发动机尾气,使用噻唑蓝比色法试剂盒检测A549细胞的相对存活率。洁净空气暴露组设为对照组,根据汽油发动机尾气中的NO_2浓度分为3个剂量组,进行剂量-效应关系分析。使用美国环境保护总署(EPA)的基准剂量工具软件(Benchmark Dose software,BMDS)中的Hill、Linear、Polynomial和Power 4种模型,根据模型拟合的AIC系数(Akaike Information Coefficient)和P值确定模型的拟合效果来计算汽油发动机尾气中NO_2的毒作用基准剂量和基准剂量下限值。结果洁净空气暴露组的NO_2浓度为0 mg/m3,A549细胞的相对存活率为90.58%±5.11%。在怠速(冷启动)、100%和53%负荷下,汽油发动机尾气中NO_2的平均浓度分别为6.6、8.2和9.2 mg/m3,A549细胞的相对存活率分别为74.27%±4.05%、63.86%±2.58%和58.77%±0.81%。Hill、Linear、Polynomial和Power 4种模型显示,随着汽油发动机尾气中NO_2浓度的升高,A549细胞的相对存活率随之降低。以拟合效果最好的Power模型估算得到的不同工况下汽油发动机尾气中NO_2的毒作用基准剂量和基准剂量下限值分别为2.83和1.96 mg/m^3。结论不同工况下汽油发动机尾气中的NO_2可对A549细胞的相对存活率产生影响,引起细胞存活率下降,并有剂量-效应关系。气-液界面暴露于汽油发动机尾气NO_2的A549细胞存活率明显降低的参考剂量为1.96 mg/m^3。Objective To explore the dose-effect relationship between the NO2 in gasoline engine exhaust(GEE） at different load conditions and the cell relative viability(CRV） of human lung cancer A549 cells by air-liquid interface(ALI）,and its benchmark dose(BMD）. Further to provide basic data for the toxic effects assessment of NO2 in automobile exhaust. Methods The GEE was collected with an air bag at idle(cold start）,and under 53% and 100% loads,respectively. The concentration of NO2 in the GEE was determined using N-(1-naphthyl）-ethylenediamine dihydrochloride spectrophotometric method. After exposed to the GEE by ALI,the CRV of human lung cancer A549 cells was analyzed by MTT assay kit. The clean air exposure group was look as the control group. According to the concentration of NO2 in the GEE,3 dose groups were divided and used to analyze the dose-effect relationship. USEPA BMDS software(version 2. 6. 0. 1）was applied to calculate the BMD and benchmark dose lower limit(BMDL） including four models(Hill,Linear,Polynomial and Power）.Finally,the BMD and BMDL for the cytotoxicity of NO2 in GEE was established according the Akaike Information Coefficient(AIC） and P value. Results The concentration of NO2 in the clean air exposed group was 0 mg/m3,and the CRV of A549 cells was 90. 58% ± 5. 11%.In the idle(cold start）,100% and 53% load exposed group,the average concentration of NO2 in the GEE were 6. 6,8. 2 and 9. 2 mg/m3,respectively,and the CRV of A549 cells were 74. 27% ± 4. 05%,63. 86% ± 2. 58% and 58. 77% ± 0. 81%,respectively. The four models including Hill,Linear,Polynomial and Power showed that the CRV of A549 cells decreased with the increase of NO2 concentration in GEE.The cytotoxicity of NO2 in GEE was estimated by the best fitting Power model,and the BMD and BMDL were 2. 83 and 1. 96 mg/m3,respectively. Conclusion The NO2 in the GEE can affect the CRV of A549 cells under different working conditions,which can lead to a dose-dependent change in the cytotoxicity. Exposed to the NO2 in the G

关 键 词：汽油发动机尾气 气-液界面 二氧化氮 基准剂量 剂量-效应关系 

分 类 号：R114[医药卫生—卫生毒理学] R135.1[医药卫生—公共卫生与预防医学]