Environmental Science and Pollution Research

, Volume 24, Issue 36, pp 27843–27854 | Cite as

NO2 inhalation enhances asthma susceptibility in a rat model

  • Ming HanEmail author
  • Xiaotong Ji
  • Guangke LiEmail author
  • Nan Sang
Research Article


Nitrogen dioxide (NO2) is a major air pollutant. Epidemiologic studies have found that NO2 exposure is associated with an increased risk of asthma. Nevertheless, the potential molecular mechanisms remain unclear. In this study, we investigated the effect of NO2 inhalation on the occurrence of allergic airway inflammation and its underlying mechanisms. Firstly, male Wistar rats were exposed to 2 and 5 mg/m3 NO2 (28 days, 5 h/day). The results showed that NO2 exposure could induce pulmonary inflammatory response, mucus formation, and Th1/Th2 imbalance in the lung of normal rats, resulting in allergic asthma-like features. Secondly, male Wistar rats were exposed to 5 mg/m3 NO2 (42 days, 5 h/day), sensitized with ovalbumin (OVA), challenged with aerosolized OVA, and characterized in asthma models. Results showed that NO2 exposure aggravated lung inflammation in the OVA-sensitized rats, accompanied by the increase in inflammatory cell infiltration, mucus hypersecretion, and collagen deposition. Furthermore, NO2 exposure promoted the increase in the expression of mucin gene (MUC5AC) and pro-inflammatory factors [interleukin (IL)-1β, intercellular adhesion molecule-1 (ICAM-1), and IL-6] as well as serum OVA-specific immunoglobulin E (IgE) production. Taken together, we established that NO2 exposure promotes allergic airway inflammation and increases the asthma susceptibility. The underlying mechanisms involve the promotion of activation of interleukin-4/signal transducer and activator of transcription-6 (IL-4/STAT6) pathway [IL-4 receptor (IL-4R) α, janus kinase (JAK) 1, JAK 3, and STAT6] and related transcription factor [T cell-specific protein-tyrosine kinase (Lck), extracellular-regulated kinase (ERK)1/2, and nuclear factor-κB (NF-κB)]. In particular, the imbalance of Th1/Th2 cell differentiation [IL-4, interferon (IFN)-γ, GATA-binding protein-3 (GATA-3), and T-box expressed in T cells (T-bet)] plays a pivotal role in NO2-induced inflammatory responses. These findings may provide a better understanding of mechanism of NO2-associated respiratory diseases.


Nitrogen dioxide Airway inflammation Allergic asthma Th1/Th2 differentiation IL-4/STAT6 pathway 


Funding information

This study was supported by the National Natural Science Foundation of PR China (NSFC, No. 21477070, 21377076, 21222701), Specialized Research Fund for the Doctoral Program of Higher Education (SRFDP, No. 20121401110003, 20131401110005), and Natural Science Foundation of Shanxi Province (No. 2015021040).


  1. Afkarian M, Sedy JR, Yang J, Jacobson NG, Cereb N, Yang SY, Murphy TL, Murphy KM (2002) T-bet is a STAT1-induced regulator of IL-12R expression in naive CD4+ T cells. Nat Immunol 3:549–557CrossRefGoogle Scholar
  2. Akira S (1999) Functional roles of STAT family proteins: lessons from knockout mice. Stem Cells 17:138–146CrossRefGoogle Scholar
  3. Ashino S, Takeda K, Li H, Taylor V, Joetham A, Pine PR, Gelfand EW (2014) Janus kinase1/3 signaling pathways are key initiators of TH2 differentiation and lung allergic responses. J Allergy Clin Immunol 133:1162–1174CrossRefGoogle Scholar
  4. Barck C, Lundahl J, Hallden G, Bylin G (2005) Brief exposures to NO2 augment the allergic inflammation in asthmatics. Environ Res 97:58–66CrossRefGoogle Scholar
  5. Barnes PJ (2008a) Immunology of asthma and chronic obstructive pulmonary disease. Nat Rev Immunol 8:183–192CrossRefGoogle Scholar
  6. Barnes PJ (2008b) The cytokine network in asthma and chronic obstructive pulmonary disease. J Clin Invest 118:3546–3556CrossRefGoogle Scholar
  7. Bascom R (1996) Health effects of outdoor air pollution. Part 2. Committee of the Environmental and Occupational Health Assembly of the American Thoracic Society. Am J Respir Crit Care Med 153:477–498CrossRefGoogle Scholar
  8. Bevelander M, Mayette J, Whittaker LA, Paveglio SA, Jones CC, Robbins J, Hemenway D, Akira S, Uematsu S, Poynter ME (2007) Nitrogen dioxide promotes allergic sensitization to inhaled antigen. J Immunol 179:3680–3688CrossRefGoogle Scholar
  9. Chakir H, Wang H, Lefebvre DE, Webb J, Scott FW (2003) T-bet/GATA-3 ratio as a measure of the Th1/Th2 cytokine profile in mixed cell populations: predominant role of GATA-3. J Immunol Meth 278:157–169CrossRefGoogle Scholar
  10. Chapman RW, Curran AK, House A, Richard J, Salisbury B, Hunter JC, Anthes JC, Phillips JE (2011) Effect of mometasone furoate (MF)/formoterol fumarate (F) combination (MF/F) on late-phase responses in allergen-challenged Brown Norway rats. Pulm Pharmacol Ther 24:67–73CrossRefGoogle Scholar
  11. Chauhan AJ, Johnston SL (2003) Air pollution and infection in respiratory illness. Br Med Bull 68:95–112CrossRefGoogle Scholar
  12. Chauhan AJ, Krishna MT, Frew AJ, Holgate ST (1998) Exposure to nitrogen dioxide (NO2) and respiratory disease risk. Rev Environ Health 13:73–90Google Scholar
  13. Choi IW, Kim DK, Ko HM, Lee HK (2004) Administration of antisense phosphorothioate oligonucleotide to the p65 subunit of NF-κB inhibits established asthmatic reaction in mice. Int Immunopharmacol 4:1817–1828CrossRefGoogle Scholar
  14. Cohn L, Homer RJ, MacLeod H, Mohrs M, Brombacher F, Bottomly K (1999) Th2-induced airway mucus production is dependent on IL-4Ralpha, but not on eosinophils. J Immunol 162:6178–6183Google Scholar
  15. Daniel C, Salvekar A, Schindler U (2000) A gain-of-function mutation in STAT6. J Biol Chem 275:14255–14259CrossRefGoogle Scholar
  16. Finotto S, Neurath MF, Glickman JN, Qin S, Lehr HA, Green FH, Ackerman K, Haley K, Galle PR, Szabo SJ, Drazen JM, De Sanctis GT, Glimcher LH (2002) Development of spontaneous airway changes consistent with human asthma in mice lacking T-bet. Science 295:336–338CrossRefGoogle Scholar
  17. Foster PS, Mould AW, Yang M, Mackenzie J, Mattes J, Hogan SP, Mahalingam S, Mckenzie AN, Rothenberg ME, Young IG, Matthaei KI, Webb DC (2001) Elemental signals regulating eosinophil accumulation in the lung. Immunol Rev 179:173–181CrossRefGoogle Scholar
  18. Grunig G, Warnock M, Wakil AE, Venkayya R, Brombacher F, Rennick DM, Sheppard D, Mohrs M, Donaldson DD, Locksley RM, Corry DB (1998) Requirement for IL-13 independently of IL-4 in experimental asthma. Science 282:2261–2263CrossRefGoogle Scholar
  19. Guo HW, Yun CX, Hou GH, Du J, Huang X, Lu Y, Keller ET, Zhang J, Deng JG (2014) Mangiferin attenuates Th1/Th2 cytokine imbalance in an ovalbumin-induced asthmatic mouse model. PLoS One 9: e100394Google Scholar
  20. Han M, Guo Z, Li GK, Sang N (2013) Nitrogen dioxide inhalation induces genotoxicity in rats. Chemosphere 90:2737–2742CrossRefGoogle Scholar
  21. Hart LA, Krishnan VL, Adcock IM, Barnes PJ, Chung KF (1998) Activation and localization of transcription factor, nuclear factor-kB, in asthma. Am J Respir Crit Care Med 158:1585–1592CrossRefGoogle Scholar
  22. Hedberg K, Hedberg CW, Iber C, White KE, Osterholm MT, Jones DB, Flink JR, MacDonald KL (1989) An outbreak of nitrogen dioxideinduced respiratory illness among ice hockey players. JAMA 262:3014–3017CrossRefGoogle Scholar
  23. Hirahara K, Yamashita M, Iwamura C, Shinoda K, Hasegawa A, Yoshizawa H, Koseki H, Gejyo F, Nakayama T (2008) Repressor of GATA regulates TH2-driven allergic airway inflammation and airway hyperresponsiveness. J Allergy Clin Immunol 122:512–520CrossRefGoogle Scholar
  24. Ho IC, Glimcher LH (2002) Transcription: tantalizing times for T cells. Cell 109:S109–S120CrossRefGoogle Scholar
  25. Hussain I, Jain VV, O’Shaughnessy P, Businga TR, Kline J (2004) Effect of nitrogen dioxide exposure on allergic asthma in a murine model. Chest 126:198–204CrossRefGoogle Scholar
  26. Ji NF, Xie YC, Zhang MS, Zhao X, Cheng H, Wang H, Yin KS, Huang M (2014) Ligustrazine corrects Th1/Th2 and Treg/Th17 imbalance in a mouse asthma model. Int Immunopharmacol 21:76–81CrossRefGoogle Scholar
  27. Ji XT, Han M, Yun Y, Li GK, Sang N (2015) Acute nitrogen dioxide (NO2) exposure enhances airway inflammation via modulating Th1/Th2 differentiation and activating JAK-STAT pathway. Chemosphere 120:722–728CrossRefGoogle Scholar
  28. Kemp KL, Levin SD, Bryce PJ, Stein PL (2010) Lck mediates Th2 differentiation through effects on T-bet and GATA-3. J Immunol 184:4178–4184CrossRefGoogle Scholar
  29. Kim MS, Cho KA, Cho YJ, Woo SY (2013) Effects of interleukin-9 blockade on chronic airway inflammation in murine asthma models. Allergy Asthma Immunol Res 5:197–206CrossRefGoogle Scholar
  30. Krishna MT, Chauhan AJ, Frew AJ, Holgate ST (1998) Toxicological mechanisms underlying oxidant pollutant-induced airway injury. Rev Environ Health 13:59–71Google Scholar
  31. Kumie A, Emmelin A, Wahlberg S, Berhane Y, Ali A, Mekonen E, Worku A, Brandstrom D (2009) Sources of variation for indoor nitrogen dioxide in rural residences of Ethiopia. Environ Health 8:51CrossRefGoogle Scholar
  32. Kuo ML, Huang JL, Yeh KW, Li PS, Hsieh KH (2001) Evaluation of Th1/Th2 ratio and cytokine production profile during acute exacerbation and convalescence in asthmatic children. Ann Allergy Asthma Immunol 86:272–276CrossRefGoogle Scholar
  33. Kuperman D, Schofield B, Wills-Karp M, Grusby MJ (1998) Signal transducer and activator of transcription factor 6 (Stat6)-deficient mice are protected from antigen-induced airway hyperresponsiveness and mucus production. J Exp Med 187:939–948CrossRefGoogle Scholar
  34. Lim SM, Kang GD, Jeong JJ, Choi HS, Kim DH (2016) Neomangiferin modulates the Th17/Treg balance and ameliorates colitis in mice. Phytomedicine 23:131–140CrossRefGoogle Scholar
  35. Liu J, Ballaney M, Al-alem U, Quan C, Jin X, Perera F, Chen LC, Miller RL (2008) Combined inhaled diesel exhaust particles and allergen exposure alter methylation of T helper genes and IgE production in vivo. Toxicol Sci 102:76–81CrossRefGoogle Scholar
  36. Liu L, Poon R, Chen L, Frescura AM, Montuschi P, Ciabattoni G, Wheeler A, Dales R (2009) Acute effects of air pollution on pulmonary function, airway inflammation, and oxidative stress in asthmatic children. Environ Health Perspect 117:668–674CrossRefGoogle Scholar
  37. Oberholzer HM, Pretorius E (2009) Investigating lung remodeling in Modul8-treated BALB/c asthmatic animals. Micron 40:775–782CrossRefGoogle Scholar
  38. Oh CK, Geba GP, Molfino N (2010) Investigational therapeutics targeting the IL-4/IL-13/STAT-6 pathway for the treatment of asthma. Eur Respir Rev 19:46–54CrossRefGoogle Scholar
  39. Ordonez CL, Khashayar R, Wong HH, Ferrando R, Wu R, Hyde DM, Hotchkiss JA, Zhang Y, Novikov A, Dolganov G, Fahy JV (2001) Mild and moderate asthma is associated with airway goblet cell hyperplasia and abnormalities in mucin gene expression. Am J Respir Crit Care Med 163:517–523CrossRefGoogle Scholar
  40. Pathmanathan S, Krishna MT, Blomberg A, Helleday R, Kelly FJ, Sandstrom T, Holgate ST, Wilson SJ, Frew AJ (2003) Repeated daily exposure to 2 ppm nitrogen dioxide upregulates the expression of IL-5, IL-10, IL-13, and ICAM-1 in the bronchial epithelium of healthy human airways. Occup Environ Med 60:892–896CrossRefGoogle Scholar
  41. Poynter ME, Persinger RL, Irvin CG, Butnor KJ, van Hirtum H, Blay W, Heintz NH, Robbins J, Hemenway D, Taatjes DJ, Janssen-Heininger Y (2006) Nitrogen dioxide enhances allergic airway inflammation and hyperresponsiveness in the mouse. Am J Physiol Lung Cell Mol Physiol 290:L144–L152CrossRefGoogle Scholar
  42. Proust B, Lacroix G, Robidel F, Marliere M, Lecomte A, Vargaftig BB (2002) Interference of a short-term exposure to nitrogen dioxide with allergic airways responses to allergenic challenges in BALB/c mice. Mediat Inflamm 11:251–260CrossRefGoogle Scholar
  43. Santus P, Russo A, Madonini E, Allegra L, Blasi F, Centanni S, Miadonna A, Schiraldi G, Amaducci S (2012) How air pollution influences clinical management of respiratory diseases. A case-crossover study in Milan. Respir Res 13:95CrossRefGoogle Scholar
  44. Shi YH, Shi GC, Wan HY, Jiang LH, Ai XY, Zhu HX, Tang W, Ma JY, Jin XY, Zhang BY (2011) Coexistence of Th1/Th2 and Th17/Treg imbalances in patients with allergic asthma. Chin Med J 124:1951–1956Google Scholar
  45. Svendsen ER, Gonzales M, Mukerjee S, Smith L, Ross M, Walsh D, Rhoney S, Andrews G, Ozkaynak H, Neas LM (2012) GIS-modeled indicators of traffic-related air pollutants and adverse pulmonary health among children in El Paso, Texas. Am J Epidemiol 176:S131–S141CrossRefGoogle Scholar
  46. Szabo SJ, Kim ST, Costa GL, Zhang X, Fathman CG, Glimcher LH (2000) A novel transcription factor, T-bet, directs Th1 lineage commitment. Cell 100:655–669CrossRefGoogle Scholar
  47. Touloumi G, Katsouyanni K, Zmirou D, Schwartz J, Spix C, de Leon AP, Tobias A, Quennel P, Rabczenko D, Bacharova L, Bisanti L, Vonk JM, Ponka A (1997) Short-term effects of ambient oxidant exposure on mortality: a combined analysis within the APHEA project. Air pollution and health: a European approach. Am J Epidemiol 146:177–185CrossRefGoogle Scholar
  48. Wan G, Wei B (2015) Erythropoietin regulates Treg cells in asthma through TGF-β receptor signaling. Am J Transl Res 7:2305–2315Google Scholar
  49. Yamashita M, Shinnakasu R, Asou H, Kimura M, Hasegawa A, Hashimoto K, Hatano N, Ogata M, Nakayama T (2005) Ras-ERK MAPK cascade regulates GATA3 stability and Th2 differentiation through ubiquitinproteasome pathway. J Biol Chem 280:29409–29419CrossRefGoogle Scholar
  50. Yeatts KB, El-Sadig M, Leith D, Kalsbeek W, Al-Maskari F, Couper D, Funk WE, Zoubeidi T, Chan RL, Trent CB, Davidson CA, Boundy MG, Kassab MM, Hasan MY, Rusyn I, Gibson JM, Olshan AF (2012) Indoor air pollutants and health in the United Arab Emirates. Environ Health Perspect 120:687–694CrossRefGoogle Scholar
  51. Zhao BT, YX S, He SS, Zhong M, Cui G (2016) Evolution and comparative assessment of ambient air quality standards in China. J Integr Environ Sci 13:85–102Google Scholar
  52. Zhu J, Min B, Hu-Li J, Watson CJ, Grinberg A, Wang Q, Killeen N, Urban JF Jr, Guo L, Paul WE (2004) Conditional deletion of Gata3 shows its essential function in T(H)1-T(H)2 responses. Nat Immunol 5:1157–1165CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany 2017

Authors and Affiliations

  1. 1.College of Environment and Resource, Research Center of Environment and HealthShanxi UniversityTaiyuanPeople’s Republic of China

Personalised recommendations