程志斌;杨峥;刘艳菊;郭青云;朱明淏;钟震宇;李俊芳;单云芳;张鹏骞;刘田;

• 1:北京麋鹿生态实验中心

摘要(Abstract)：

机动车尾气是我国大气细颗粒物PM_(2.5)污染的主要来源之一,长期暴露于机动车尾气对动物呼吸系统及其他多种脏器造成损害。为探索长期暴露于汽油燃烧的汽车尾气对小鼠肺组织的损伤作用,将51只雄性8周龄SPF级ICR小鼠随机分为7个处理。其中,空气对照组1个(6只);箱体对照组3个:0.5 h对照组(5只)、1 h对照组(8只)、2 h对照组(8只);汽油车尾气暴露组3个:0.5 h暴露组(8只)、1 h暴露组(8只)、2 h暴露组(8只)。小鼠每天在自制染毒箱中暴露0.5 h、1 h、2 h。95 d的暴露实验后,处死实验鼠,测定其肺组织的炎症因子白介素-6(IL-6)、乳酸脱氢酶(LDH)、肿瘤坏死因子-α(TNF-α)、转化生长因子β(TGF-β),并制作小鼠肺部病理学切片进行HE染色。暴露实验过程中,观察记录小鼠行为特征,测定箱体内O_2、NO_x、NO_2、NO、SO_2和CO的浓度。采集汽油车尾气的PM2.5,对其水溶性阴离子(F~-、Cl~-、NO_2~-、NO_3~-、SO_4~(2-))、水溶性阳离子(Na~+、NH_4~+、Ca~(2+)、K~+、Mg~(2+))、有机碳(OC)和元素碳(EC)、7种重金属元素(Cu、Ni、Mn、Zn、Pb、As、Cr)、16种多环芳烃(PAHs)浓度等进行了测定。结果表明:(1)亚慢性染毒箱内汽油车尾气PM2.5所分析的化学物质中,OC浓度最高,占PM2.5中总化学物质浓度的75.5%;其次是EC,占22.9%;再次是阴离子和阳离子,分别占1.11%和0.27%。(2)3个暴露组小鼠活跃程度降低,暴露5～8 min后进入睡眠或休息状态,呼吸频率降低,小鼠通过这些行为来减少汽油车尾气的伤害。(3)汽油车尾气长期暴露组的体重增长率与各自对照组之间不存在显著差异,但2 h暴露组体重增长率明显小于2 h对照组,且2 h暴露组在第14周开始体重有下降的趋势。(4)与对照组相比,1 h暴露组的IL-6和0.5 h暴露组的TNF-α显著升高,说明汽油车尾气暴露造成小鼠肺炎症损伤,使IL-6、TNF-α的分泌增加。随着暴露时间的延长,IL-6和TNF-α表现为2 h暴露组分别显著低于1 h暴露组和0.5 h暴露组,意味长期暴露后小鼠肺部受到严重伤害,免疫力严重下降,不再有积极的抗炎性反应。(5)组织病理观察表明,与开放对照组比较,3个汽油车尾气暴露组小鼠肺部组织充血较为严重,大量淋巴细胞、中性粒细胞浸润,肺泡间质增生明显,呈现出随时间延长损伤加重的梯度差异。(6)汽油车尾气长期暴露可致实验鼠肺脏损伤,且随着暴露时间的增加,对肺组织造成的损伤程度增加。

关键词(KeyWords)： 汽油车尾气;颗粒物;小鼠;化学成分;染毒箱;炎症因子;病理伤害

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金(41475133)

作者(Author): 程志斌;杨峥;刘艳菊;郭青云;朱明淏;钟震宇;李俊芳;单云芳;张鹏骞;刘田;

Email:

DOI:

参考文献(References)：

• [1] Lang J, Cheng S, Wei W, et al. A study on the trends of vehicular emissions in the Beijing–Tianjin–Hebei(BTH)Region, China[J]. Atmospheric Environment,2012, 62:605-614
• [2] Wang Y S, Yao L, Wang L L, et al. Mechanism for the formation of the January 2013 heavy haze pollution episode over central and eastern China[J]. Science China:Earth Sciences, 2014, 44(1):14-25
• [3] Goel R, Guttikunda S K. Evolution of on-road vehicle exhaust emissions in Delhi[J]. Atmospheric Environment,2015, 105:78-90
• [4] Amato F, Alastuey A, Karanasiou A, et al. AIRUSE-LIFE+:A harmonized PM speciation and source apportionment in five southern European cities[J]. Atmospheric Chemistry and Physics, 2016, 16(5):3289-3309
• [5]韩力慧,张鹏,张海亮,等.北京市大气细颗粒物污染与来源解析研究[J].中国环境科学, 2016, 36(11):3203-3210Han L H, Zhang P, Zhang H L, et al. Pollution and source apportionment of atmospheric fine particles in Beijing[J].China Environmental Science, 2016, 36(11):3203-3210(in Chinese)
• [6]杨昆昊,夏赞宇,何芃,等.机动车燃油质量及尾气排放与北京市大气污染的相关性[J].中国科学院大学学报, 2017, 34(3):304-317Yang K H, Xia Z Y, He P, et al. Correlation of fuel quality and emissions of motor vehicle with atmospheric pollution in Beijing[J]. Journal of University of Chinese Academy of Sciences, 2017, 34(3):304-317(in Chinese)
• [7]薛国强,朱彬,王红磊.南京市大气颗粒物中水溶性离子的粒径分布和来源解析[J].环境科学, 2014, 35(5):1633-1643Xue G Q, Zhu B, Wang H L. Size distributions and source apportionment of soluble ions in aerosol in Nanjing[J].Environmental Science, 2014, 35(5):1633-1643(in Chinese)
• [8] Kagawa J. Health effects of diesel exhaust emissions—A mixture of air pollutants of worldwide concern[J]. Toxicology, 2002, 181:349-353
• [9] Rengaraj D, Kwon W S, Pang M G. Effects of motor vehicle exhaust on male reproductive function and associated proteins[J]. Journal of Proteome Research, 2014, 14(1):22-37
• [10] Reis H, Reis C, Sharip A, et al. Diesel exhaust exposure,its multi-system effects, and the effect of new technology diesel exhaust[J]. Environment International, 2018, 114:252-265
• [11]邵瑜,程义斌,徐东群,等.机动车尾气暴露与交通协管员呼吸系统症状和肺功能的关系研究[J].环境与健康杂志, 2010, 27(5):436-439Shao Y, Cheng Y B, Xu D Q. Study on relationship between automobile exhaust exposure and respiratory syndromes and lung function among traffic police assistants[J]. Journal of Environment and Health, 2010, 27(5):436-439(in Chinese)
• [12]王少利,郭新彪,张金良.北京市大气污染对学龄儿童呼吸系统疾病和症状的影响[J].环境与健康杂志,2004, 21(1):41-44Wang S L, Guo X B. Study on the effects of ambient air pollution on respiratory disease and symptoms among school-age children in Beijing[J]. Journal of Environment and Health, 2004, 21(1):41-44(in Chinese)
• [13]刘美娟,董光辉,潘国伟,等.鞍山市大气污染对儿童呼吸系统健康的影响[J].环境与健康杂志, 2006, 23(3):198-201Liu M J, Dong G H, Pan G W. Effects of air pollution on respiratory system of children in Anshan City[J]. Journal of Environment and Health, 2006, 23(3):198-201(in Chinese)
• [14] Benbrahim-Tallaa L, Baan R A, Grosse Y, et al. Carcinogenicity of diesel-engine and gasoline-engine exhausts and some nitroarenes[J]. The lancet Oncology, 2012, 13(7):663-664
• [15]刘奇琛,黄婧,郭新彪.北京市大气挥发性有机物(VOCs)的污染特征及来源[J].生态毒理学报, 2017, 12(3):49-61Liu Q C, Huang J, Guo X B. Pollution characteristics and sources of ambient volatile organic compounds(VOCs)in Beijing[J]. Asian Journal of Ecotoxicology, 2017, 12(3):49-61(in Chinese)
• [16] Widziewicz K, Rogula-Kozowska W, Rogula-Kopiec P,et al. PM1 and PM1-bound metals during dry and wet periods:Ambient concentration and health effects[J]. Environmental Engineering Science, 2017, 34(5):312-320
• [17] Künzli N, Kaiser R, Medina S, et al. Public-health impact of outdoor and traffic-related air pollution:A European assessment[J]. The Lancet, 2000, 356(9232):795-801
• [18]张遵真,吴媚,张浩,等.汽油车尾气对人肺腺癌A549细胞的氧化损伤效应研究[J].生态毒理学报, 2006, 1(1):25-29Zhang Z Z, Wu M, Zhang H, et al. Oxidative damage induced by extract from gasoline-fueled vehicle emissions in human lung adenocarcinoma A549 cells[J]. Asian Journal of Ecotoxicology, 2006, 1(1):25-29(in Chinese)
• [19]徐雯,江俊康,杨北平,等.不同粒径汽车尾气颗粒物对A549细胞毒性作用的比较[J].环境科学学报, 2013,33(12):3407-3412Xu W, Jiang J K, Yang B P, et al. Comparison of the toxic effects in A549 cells induced by automobile exhaust particulates with different sizes[J]. Acta Scientiae Circumstantiae, 2013, 33(12):3407-3412(in Chinese)
• [20]车望军,王玲,罗擎英,等.汽油车尾气致大鼠肺组织氧化损伤机制的研究[J].四川大学学报:医学版, 2009,40(1):11-14Che W J, Wang L, Luo Q Y, et al. Oxidative damage of gasoline engine exhausts to rat lung tissues[J]. Journal of Sichuan University:Medical Science Edition, 2009, 40(1):11-14(in Chinese)
• [21] Nagashima M, Kasai H, Yokota J, et al. Formation of an oxidative DNA damage, 8-hydroxydeoxyguanosine, in mouse lung DNA after intratracheal instillation of diesel exhaust particles and effects of high dietary fat and betacarotene on this process[J]. Carcinogenesis, 1995, 16(6):1441-1445
• [22]车望军,吴媚,张遵真,等.汽油车尾气对大鼠睾丸组织的氧化损伤和遗传毒性作用[J].卫生研究, 2008, 37(4):417-420Che W J, Wu M, Zhang Z Z, et al. Effects of the extracts of condensate, particulates and semivolatile organic compounds from gasoline exhaust on oxidative damage and genotoxicity in testicles of rats[J]. Journal of Hygiene Research, 2008, 37(4):417-420(in Chinese)
• [23]石鑫,陈倩,李婷华,等.吸入汽车尾气对小鼠肺部菌群的影响[J].上海交通大学学报:医学版, 2018, 38(2):146-150Shi X, Chen Q, Li T H, et al. Effects of inhaling vehicle exhaust on lung microbiome in mice[J]. Journal of Shanghai Jiaotong University, 2018, 38(2):146-150(in Chinese)
• [24]王晔,刘妍彤,薛灵抒,等.汽油车尾气致慢性肺损伤的形态学改变研究[J].现代预防医学, 2012, 39(6):1349-1354Wang Y, Liu Y T, Xue L S, et al. Study on the morphological changes of the rats’lung caused by exhausting of gasoline engine[J]. Modern Preventive Medicine, 2012,39(6):1349-1354(in Chinese)
• [25]王海青,成要平,何玉洁.妊娠期大鼠汽车尾气暴露对仔鼠生殖毒性及遗传毒性研究[J].环境与健康杂志,2011, 28(2):122-124Wang H Q, Cheng Y P, He Y J. Productive toxicity and genotoxicity of automobile exhaust[J]. Journal of Environment and Health, 2011, 28(2):122-124(in Chinese)
• [26] Liu Q Y, Liu Y J, Yin J, et al. Chemical characteristics and source apportionment of PM10, during Asian dust storm and non-dust storm days in Beijing[J]. Atmospheric Environment, 2014, 91(1):85-94
• [27] Liu Y J, Zhang T T, Liu Q Y, et al. Seasonal variation of physical and chemical properties in TSP, PM10and PM2.5at a roadside site in Beijing and their influence on atmospheric visibility[J]. Aerosol and Air Quality Research,2014, 14:954-969
• [28]王欣欣,刘庆阳,刘艳菊,等.二级热脱附-气相色谱-质谱联用测定大气可吸入颗粒物中的16种多环芳烃[J].色谱, 2010, 28(9):849-853Wang X X, Liu Q Y, Liu Y J, et al. Determination of 16polycyclic aromatic hydrocarbons in airborne particle samples using two-step thermal desorption and gas chromatography-mass spectrometry[J]. Chinese Journal of Chromatography, 2010, 28(9):849-853(in Chinese)
• [29]程志斌,刘艳菊,郭青云,等.沙尘暴源区土壤盐碱扬尘对大鼠呼吸系统的免疫损伤[J].生态毒理学报,2018, 13(2):127-138Cheng Z B, Liu Y J, Guo Q Y, et al. Immunologic injuries on respiratory system of rats induced by saline-alkali soil dust from dust storm source region[J]. Asian Journal of Ecotoxicology, 2018, 13(2):127-138(in Chinese)
• [30]程志斌,刘艳菊,亓学奎,等.北京不同污染天气PM2.5和PM10染毒对大鼠生理病理参数的影响[J].生态毒理学报, 2018, 13(3):226-240Cheng Z B, Liu Y J, Qi X K, et al. Influence of PM2.5and PM10exposure on physiological and pathological parameters of rats under different polluted weather in Beijing[J].Asian Journal of Ecotoxicology, 2018, 13(3):226-240(in Chinese)
• [31]刘晓莉,陈巍,乔德才.二氧化硫污染对运动大鼠血清和肺组织IL-6、TNF-α水平的影响[J].中国运动医学杂志, 2009, 28(1):51-54Liu X L, Chen W, Qiao D C. Effect of sulfur dioxide pollution on IL-6 and TNF-αlevel in serum and lungs in exercising rats[J]. Chinese Journal of Sports Medicine,2009, 28(1):51-54(in Chinese)
• [32] Giles L V, Barn P, Künzli N, et al. From good intentions to proven interventions:Effectiveness of actions to reduce the health impacts of air pollution[J]. Environmental Health Perspectives, 2011, 119(1):29-36
• [33] Ban J, Zhou L, Zhang Y, et al. The health policy implications of individual adaptive behavior responses to smog pollution in urban China[J]. Environment International,2017, 106:144-152
• [34]林健,黄宗锈,林蔚,等. SD大鼠体重和主要脏器正常参考值探讨及相关分析[J].海峡预防医学杂志, 2005,11(5):24-26Lin J, Huang Z X, Lin W. Study on the normal reference value and correlation analysis of body weight and the main organs of SD rat[J]. Strait Journal of Preventive Medicine, 2005, 11(5):24-26(in Chinese)
• [35]严超,曹希宁,沈炼桔,等.汽车尾气来源PM2.5长期暴露导致大鼠多器官损害[J].重庆医科大学学报, 2015(6):844-849Yan C, Cao X N, Shen L J, et al. Long exposure to automobile exhaust source PM2.5leads to inflammatory injury of multiple organs in SD rats[J]. Journal of Chongqing Medical University, 2015(6):844-849(in Chinese)
• [36]武辉.五龙颗粒对汽车尾气所致大鼠免疫抑制的作用研究[D].成都:成都中医药大学, 2004:35-36Wu H. Effect of Wulong prescription on immunosuppression induced by automobile exhaust in rats[D]. Chengdu:Chengdu University of TCM, 2004:35-36(in Chinese)
• [37]杨伟. NO-NOS信号通路介导汽车尾气暴露下致雄鼠生精功能损伤的初步研究[D].重庆:第三军医大学,2015:19-22Yang W. Effect of automobile exhaust exposure on male mice spermatogenesis function mediated by NO-NOS pathway[D]. Chongqing:Third Military Medical University, 2015:19-22(in Chinese)
• [38]张志红,杨文敏,黄铁明.汽油车尾气颗粒物对小鼠免疫功能的影响[J].中国公共卫生学报, 1999, 18(6):353-355Zhang Z H, Yang W M, Huang T M. The effect of gasoline exhaust particles on some immune functions of mouse[J]. Chinese Journal of Public Health, 1999, 18(6):353-355(in Chinese)
• [39]张澍,李晓林,林兆川,等.汽车尾气中含铂颗粒物与非含铂颗粒物肺毒性比较[J].环境与职业医学, 2006,23(2):89-92Zhang S, Li X L, Lin Z C, et al. Comparison of pulmonary toxicity induced by platinic and non-platinic particles emitted from motor vehicle exhaust[J]. Journal of Environmental&Occupational Medicine, 2006, 23(2):89-92(in Chinese)
• [40]张帆,刘健,端淑杰.白介素37的生物学特性及其与疾病的相关性研究进展[J].中国临床保健杂志, 2018, 21(1):135-138Zhang F, Liu J, Duan S J. Research progress of relationship between interleukin-37 and diseases[J]. Chinese Journal of Clinical Healthcare, 2018, 21(1):135-138(in Chinese)
• [41]吕建媛,李宏芬,宋海澄,等. TNF-α在强直性脊柱炎患者血清中的表达[J].标记免疫分析与临床, 2014, 21(6):786-787Lv J Y, Li H F, Song H C, et al. Expression of TNF-αin serum of patients with ankylosing spondylitis[J]. Labeled Immunoassays and Clinical Medicine, 2014, 21(6):786-787(in Chinese)
• [42]刘镕,赵琴平,董惠芬,等. TGF-β信号传导通路及其生物学功能[J].中国病原生物学杂志, 2014(1):77-83Liu R, Zhao Q P, Dong H F, et al. The TGF-βsignaling pathways and their biological functions[J]. Journal of Pathogen Biology, 2014(1):77-83(in Chinese)
• [43]谭强,宋宏,陈穗梅,等.机动车尾气污染对学龄儿童肺功能及炎症因子水平的影响[J].环境与健康杂志,2009(9):766-769Tan Q, Song H, Chen S M. Effect of vehicle exhaust pollution on lung function and inflammatory factors level in pupils[J]. Journal of Environment and Health, 2009(9):766-769(in Chinese)
• [44]李可欣,曹红英,王斌,等.三种典型纳米颗粒物造成的人体肺细胞氧化应激和炎症效应[J].生态毒理学报,2010, 5(6):787-792Li K X, Cao H Y, Wang B, et al. Oxidative stress and inflammatory effect on A549 cell line induced by three typical nano-particles[J]. Asian Journal of Ecotoxicology,2010, 5(6):787-792(in Chinese)
• [45]刘爱红,孙康宁,赵冬梅.多壁碳纳米管引起白鼠肺部毒性的评价[J].生态毒理学报, 2010, 5(5):724-729Liu A H, Sun K N, Zhao D M. Comparative pulmonary toxic assessment of mutil-wall carbon nanotubes in rats[J]. Asian Journal of Ecotoxicology, 2010, 5(5):724-729(in Chinese)
• [46] Liu Q, Baumgartner J, Zhang Y, et al. Source apportionment of Beijing air pollution during a severe winter haze event and associated pro-inflammatory responses in lung epithelial cells[J]. Atmospheric Environment, 2016, 126(2):28-35
• [47]樊磊,曹继军,张英杰,等.汽车尾气暴露对小鼠运动力竭时间的影响[J].生态毒理学报, 2017, 12(5):287-293Fan L, Cao J J, Zhang Y J, et al. The effect of vehicle exhaust exposure on the running exhausting time of mice[J]. Asian Journal of Ecotoxicology, 2017, 12(5):287-293(in Chinese)
• [48]孟紫强.环境毒理学[M].北京:中国环境科学出版杜,2000:353-358
• [49]杨凌.大气细颗粒物急性、亚慢性染毒对小鼠肺损伤及其免疫机制的研究[D].上海:复旦大学, 2014:10-40Yang L. Study on effects and immune mechanism of acute and sub-chronic exposure of PM2.5on lung in mice[D].Shanghai:Fudan University, 2014:10-40(in Chinese)
• [50] Chuang H C, Ho K F, Cao J J, et al. Effects of non-protein-type amino acids of fine particulate matter on E-cadherin and inflammatory responses in mice[J]. Toxicology Letters, 2015, 237(3):174-180
• [51]杜青平,孟紫强.二氧化硫对大鼠肺细胞膜通透性的损伤效应[J].中国公共卫生, 2003, 19(7):783-785Du Q P, Meng Z Q. Damage effects of sulfur dioxide on pulmonary membranous permeability of rats[J]. China Public Health, 2003, 19(7):783-785(in Chinese)
• [52]田锁臣,吴铁军,王可富.急性重度一氧化碳中毒大鼠肿瘤坏死因子及白细胞介素的变化[J].中华危重病急救医学, 2004, 16(12):760-761Tian S C, Wu T J, Wang K F. Changes of tumor necrosis factor and interleukin in acute severe carbon monoxide poisoning rats[J]. Chinese Critical Care Medicine, 2004,16(12):760-761(in Chinese)
• [53]李自力,康学文,郭豫学,等.纳洛酮对急性CO中毒大鼠血清肿瘤坏死因子α、白介素8含量的影响[J].中国急救复苏与灾害医学杂志, 2007, 2(12):718-719Li Z L, Kang X W, Guo Y X, et al. Serum tumor necrosis factor-αand interleukin-8 changes in acute carbon monoxide pinioning and the effect of naloxone:Experiment with rats[J]. China Journal of Emergency Resuscitation&Disaster Medicine, 2007, 2(12):718-719(in Chinese)
• [54]付双来.急性一氧化碳中毒患者血清酶学变化及其临床意义[J].现代中西医结合杂志, 2002, 11(15):1417-1418Fu S L. Changes and climical significance of serum enzymes of acute carbon monoxide poisoning[J]. Modern Journal of Integrated Traditional Chinese and Western Medicine, 2002, 11(15):1417-1418(in Chinese)
• [55]陈轶,陈志斌,詹鹃铭,等.地下停车场汽车尾气污染对小鼠血清炎症因子影响的研究[J].中国中西医结合急救杂志, 2013, 20(6):353-354Chen Y, Chen Z B, Zhan J M, et al. A study on effect of automobile exhaust pollutants in under ground parking area on serum inflammatory cytokines of mice[J]. Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care, 2013, 20(6):353-354(in Chinese)
• [56]邵丹青,杜毅鹏,冯娟,等.大气污染细颗粒中有机成分苯并芘直接引起人Ⅱ型肺上皮细胞慢性炎症[J].中国病理生理杂志, 2015(10):1920Shao D Q, Du Y P, Feng J, et al. Benzopyrene, an organic component of fine air pollution particles, directly causes chronic inflammation of human typeⅡpulmonary epithelial cells[J]. Chinese Journal of Pathophysiology, 2015(10):1920(in Chinese)
• [57] UmannováL, Machala M, Topinka J, et al. Benzo[a]pyrene and tumor necrosis factor-αcoordinately increase genotoxic damage and the production of proinflammatory mediators in alveolar epithelial type II cells[J]. Toxicology Letters, 2011, 206(2):121-129
• [58] Dreij K, Rhrissorrakrai K, Gunsalus K C, et al. Benzo[a]pyrene diol epoxide stimulates an inflammatory response in normal human lung fibroblasts through a p53 and JNK mediated pathway[J]. Carcinogenesis, 2010, 31(6):1149-1157
• [59]孟紫强,刘玉香,武冬梅.二氧化硫对小鼠肺及血清中细胞因子的影响[J].中国公共卫生, 2004, 20(9):1050-1051Meng Z Q, Liu Y X, Wu D M. Effect of sulfur dioxide inhalation on cytokine levels in lungs and serum of mice[J]. China Public Health, 2004, 20(9):1050-1051(in Chinese)
• [60]张迪.各种汽车尾气对小白鼠部分器官组织形态学上的影响[D].大连:辽宁师范大学, 2009:18-19Zhang D. Automobile exhaust on mouse tracheal study the impact of organizational form[D]. Dalian:Liaoning Normal University, 2009:18-19(in Chinese)