摩托车尾气中非甲烷总烃和氮氧化物水平与BEAS-2B细胞存活率的关系研究  

Association between the levels of non methane total hydrocarbons and nitrogen oxides and the survival rate of BEAS-2B cells exposed to motorcycle exhaust

作　　者：鱼涛[1] 崔强[2] 张妍[2] 刘浏[2] 钟磊[2] 胡晓宇[2] 李树飞[3] 李斌[1] 郑玉新[4] 宾萍[1] YU Tao;CUI Qiang;ZHANG Yan;LIU Liu;ZHONG Lei;HU Xiao-yu;LI Shu-fei;LI Bin;ZHENG Yu-xin;BIN Ping(National Institute of Occupational Health and Poison Control,Chinese Center for Disease Control and Prevention,Beijing 100050,China;Beijing Center for Disease Prevention and Control,Beijing 100013,China;Tianjin Center for Disease Prevention and Control,Tianjin 300011,China;School of Public Health,Qingdao University,Shandong Qingdao 266021,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,北京100050 [2]北京市疾病预防控制中心,北京100013 [3]天津市疾病预防控制中心,天津300011 [4]青岛大学公共卫生学院,山东青岛266021

出　　处：《毒理学杂志》2021年第5期401-404,411,共5页Journal of Toxicology

基　　金：国家自然科学基金(81472955,91643203,82073596)。

摘　　要：目的应用气—液界面(Air-liquid interface,ALI)染毒技术模拟肺部细胞暴露摩托车尾气(Motorcycle exhaust,ME)的方式,探讨ME中非甲烷总烃(Non methane total hydrocarbon,NMHC)和氮氧化物(Nitrogen oxides,NOx)水平与BEAS-2B细胞存活率的关系。方法固定一辆配备三元催化器的摩托车,用洁净空气将ME直接稀释成体积比VME∶V洁净空气的比例分别为1∶20、1∶10、1∶5和1∶2的不同配比稀释的ME;用采气袋收集不同配比稀释后的ME,气相色谱法分析NMHC的水平;用盐酸萘乙二胺分光光度法测定NOx的浓度。采用ALI染毒系统,在染毒流量为25 ml/min,37℃水浴条件下,对BEAS-2B细胞持续染毒60 min。通过CCK-8细胞毒性检测试剂盒测定细胞的相对存活率。结果随着ME配比稀释的倍数增加,尾气中NMHC和NOx的浓度呈现降低的趋势,NMHC和NOx的浓度与稀释倍数存在有统计学意义的负相关性(rSpearman分别为-0.859,-0.982,P值均为<0.001);细胞相对存活率随着ME配比稀释的倍数增加而增加,存在有统计学意义的正相关性(rSpearman为0.839,P<0.001),从1∶20稀释组到未稀释尾气组,细胞相对存活率的均数依次降低了4.71%、18.28%、26.98%、44.92%和42.59%;ME中NMHC和NOx的水平与BEAS-2B细胞的相对存活率呈负相关性,并具有统计学意义(r分别为-0.956、-0.892,P值分别为0.011、0.042)。结论ME中不同水平的NMHC和NOx可明显影响BEAS-2B细胞的相对存活率,并具有一定的剂量—效应关系。Objective To investigate the relationship between the levels of non methane total hydrocarbons(NMHC)and nitrogen oxides(NOx)and the survival rate of BEAS-2B cells,air liquid interface(ALI)was used to simulate the way of lung cells exposed to motorcycle exhaust(ME).Methods A motorcycle equipped with three-way catalytic converter was fixed.Four concentrations were included by a direct dilution method and the ME was diluted into 1∶20,1∶10,1∶5 and 1∶2 according the ratio of VME/V Clean air.All ME samples were collected using Tedlar bag.The level of NMHC was analyzed by gas chromatography.The concentration of NOx was determined using N-(1-naphthyl)-ethylenediamine dihydrochloride spectrophotometric method.Controlled by circulating water at 37℃,and a flow rate of 25 ml/min/well for 1 h,BEAS-2B cells were exposed to ME or clean air in the ALI exposure device.All of the cell relative viability(CRV)was determined using Cell Counting Kit-8(CCK-8)assay.Results With the increase of the dilution ratio of ME,the concentration of NMHC and NOx were decreased.There was a significant negative correlation between the concentration of NMHC and NOx and the dilution ratio of ME(the correlation coefficients were-0.859,-0.982,respectively,P<0.001).The CRV increased with the increase of dilution ratio of ME,and there was a significally positive correlation(correlation coefficient was 0.839,P<0.001).From the 1:20 dilution group to the non-dilution group,the mean CRV of BEAS-2B cells decreased by 4.71%,18.28%,26.98%,44.92%and 42.59%,respectively;the levels of NMHC and NOx in ME were negatively correlated with the CRV of BEAS-2B cells,and they were statistically significant(the r was-0.956 and-0.892,P=0.011 and 0.042,respectively).Conclusion Different levels of NMHC and NOx in ME can significantly affect the CRV of BEAS-2B cells,and there is a certain dose-response relationship.

关 键 词：摩托车尾气 气—液界面 非甲烷总烃 氮氧化物 细胞存活率 

分 类 号：R114[医药卫生—卫生毒理学]