Inflammation Research

, Volume 64, Issue 5, pp 363–371 | Cite as

Different profiles of notch signaling in cigarette smoke-induced pulmonary emphysema and bleomycin-induced pulmonary fibrosis

  • Shi Li
  • Xiaofei Hu
  • Zheng Wang
  • Meng Wu
  • Jinnong ZhangEmail author
Original Research Paper



Different profiles of Notch signaling mediate naive T cell differentiation which might be involved in pulmonary emphysema and fibrosis.


C57BL/6 mice were randomized into cigarette smoke (CS) exposure, bleomycin (BLM) exposure, and two separate groups of control for sham exposure to CS or BLM. The paratracheal lymph nodes of the animals were analyzed by real-time PCR and immunohistochemistry. Morphometry of the lung parenchyma, measurement of the cytokines, and cytometry of the bronchoalveolar lavage fluid (BALF) were also done accordingly.


In comparison with controls, all Notch receptors and ligands were upregulated by chronic CS exposure, especially Notch3 and DLL1 (P < 0.01), and this was in line with emphysema-like morphology and Th1-biased inflammation. While Notch3 and DLL1 were downregulated by BLM exposure (P < 0.01), those was in line with fibrotic lung remodeling and Th2 polarization.


This founding implies that the CS exposure but not the BLM exposure is capable of initiating Notch signaling in lymphoid tissue of the lung, which is likely relevant to the pathogenesis of pulmonary emphysema. Unable to initiate the Th1 response or inhibit it may lead to Th2 polarization and aberrant repair.


Notch Th1/Th2 response Pulmonary emphysema Pulmonary fibrosis 



Cigarette smoke




Excessive extracellular matrix


Delta-like ligand


Bronchoalveolar lavage fluid


Mean linear intercept


Internal surface area


Chronic obstructive pulmonary disease



This research is supported by the science foundation of Hubei Province, China (Grant No.2008CDB153). The authors thank Dr. Chen Yaobin and Dr. Xia Qin for their technique assistance.

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical standard

The protocols for the animal experiments were approved by Tongji Medical College, Huazhong University of Science and Technology, and Hubei Laboratory Animal Society (Wuhan, China).

Supplementary material

11_2015_816_MOESM1_ESM.pdf (87 kb)
Supplementary material 1 (PDF 86 kb)


  1. 1.
    Crystal RG, Bitterman PB, Mossman B, Schwarz MI, Sheppard D, Almasy L, et al. Future research directions in idiopathic pulmonary fibrosis: summary of a National Heart, Lung, and Blood Institute working group. Am J Respir Crit Care Med. 2002;166:236–46.CrossRefPubMedGoogle Scholar
  2. 2.
    Chilosi M, Poletti V, Rossi A. The pathogenesis of COPD and IPF: distinct horns of the same devil. Respir Res. 2012;13:3.CrossRefPubMedCentralPubMedGoogle Scholar
  3. 3.
    Grumelli S, Corry DB, Song LZ, Song L, Green L, Huh J, et al. An immune basis for lung parenchymal destruction in chronic obstructive pulmonary disease and emphysema. PLoS Med. 2004;1:e8.CrossRefPubMedCentralPubMedGoogle Scholar
  4. 4.
    Wang Z, Chen YW, Zhang JN, Hu XF, Peng MJ. Pentoxifylline attenuates cigarette smoke-induced overexpression of CXCR3 and IP-10 in mice. Chin Med J (Engl). 2012;125:1980–5.Google Scholar
  5. 5.
    Eppert BL, Wortham BW, Flury JL, Borchers MT. Functional characterization of T cell populations in a mouse model of chronic obstructive pulmonary disease. J Immunol. 2013;190:1331–40.CrossRefPubMedCentralPubMedGoogle Scholar
  6. 6.
    Zhang J, Zheng L, Bai M. Dynamic expression of tumor necrosis factor-alpha and vascular endothelial growth factor in rat model of pulmonary emphysema induced by smoke exposure. J Huazhong Univ Sci Technolog Med Sci. 2007;27:505–7.CrossRefPubMedGoogle Scholar
  7. 7.
    Churg A, Zhou S, Preobrazhenska O, Tai H, Wang R, Wright JL. Expression of profibrotic mediators in small airways versus parenchyma after cigarette smoke exposure. Am J Respir Cell Mol Biol. 2009;40:268–76.CrossRefPubMedGoogle Scholar
  8. 8.
    Tilley AE, Harvey BG, Heguy A, Hackett NR, Wang R, O’Connor TP, et al. Down-regulation of the notch pathway in human airway epithelium in association with smoking and chronic obstructive pulmonary disease. Am J Respir Crit Care Med. 2009;179:457–66.CrossRefPubMedCentralPubMedGoogle Scholar
  9. 9.
    Wang Z, Zhang JN, Hu XF, Chen XL, Wang XR, Zhao TT, et al. Effects of pentoxifylline on Wnt/beta-catenin signaling in mice chronically exposed to cigarette smoke. Chin Med J (Engl). 2010;123:2688–94.Google Scholar
  10. 10.
    Mitaka K, Miyazaki Y, Yasui M, Furuie M, Miyake S, Inase N, et al. Th2-biased immune responses are important in a murine model of chronic hypersensitivity pneumonitis. Int Arch Allergy Immunol. 2011;154:264–74.CrossRefPubMedGoogle Scholar
  11. 11.
    Wynn TA. Integrating mechanisms of pulmonary fibrosis. J Exp Med. 2011;208:1339–50.CrossRefPubMedCentralPubMedGoogle Scholar
  12. 12.
    Lukacs NW, Hogaboam C, Chensue SW, Blease K, Kunkel SL. Type 1/type 2 cytokine paradigm and the progression of pulmonary fibrosis. Chest. 2001;120:5S–8S.CrossRefPubMedGoogle Scholar
  13. 13.
    Shimizu Y, Kuwabara H, Ono A, Higuchi S, Hisada T, Dobashi K, et al. Intracellular Th1/Th2 balance of pulmonary CD4(+) T cells in patients with active interstitial pneumonia evaluated by serum KL-6. Immunopharmacol Immunotoxicol. 2006;28:295–304.CrossRefPubMedGoogle Scholar
  14. 14.
    Konigshoff M, Balsara N, Pfaff EM, Kramer M, Chrobak I, Seeger W, et al. Functional Wnt signaling is increased in idiopathic pulmonary fibrosis. PLoS One. 2008;3:e2142.CrossRefPubMedCentralPubMedGoogle Scholar
  15. 15.
    Konigshoff M, Kramer M, Balsara N, Wilhelm J, Amarie OV, Jahn A, et al. WNT1-inducible signaling protein-1 mediates pulmonary fibrosis in mice and is upregulated in humans with idiopathic pulmonary fibrosis. J Clin Invest. 2009;119:772–87.PubMedCentralPubMedGoogle Scholar
  16. 16.
    Wang Z, Zheng T, Zhu Z, Homer RJ, Riese RJ, Chapman HA Jr, et al. Interferon gamma induction of pulmonary emphysema in the adult murine lung. J Exp Med. 2000;192:1587–600.CrossRefPubMedCentralPubMedGoogle Scholar
  17. 17.
    Sandler NG, Mentink-Kane MM, Cheever AW, Wynn TA. Global gene expression profiles during acute pathogen-induced pulmonary inflammation reveal divergent roles for Th1 and Th2 responses in tissue repair. J Immunol. 2003;171:3655–67.CrossRefPubMedGoogle Scholar
  18. 18.
    Kimura T, Ishii Y, Yoh K, Morishima Y, Iizuka T, Kiwamoto T, et al. Overexpression of the transcription factor GATA-3 enhances the development of pulmonary fibrosis. Am J Pathol. 2006;169:96–104.CrossRefPubMedCentralPubMedGoogle Scholar
  19. 19.
    Adler SH, Chiffoleau E, Xu L, Dalton NM, Burg JM, Wells AD, et al. Notch signaling augments T cell responsiveness by enhancing CD25 expression. J Immunol. 2003;171:2896–903.CrossRefPubMedGoogle Scholar
  20. 20.
    Amsen D, Blander JM, Lee GR, Tanigaki K, Honjo T, Flavell RA. Instruction of distinct CD4 T helper cell fates by different notch ligands on antigen-presenting cells. Cell. 2004;117:515–26.CrossRefPubMedGoogle Scholar
  21. 21.
    Radtke F, Fasnacht N, Macdonald HR. Notch signaling in the immune system. Immunity. 2010;32:14–27.CrossRefPubMedGoogle Scholar
  22. 22.
    Xu K, Moghal N, Egan SE. Notch signaling in lung development and disease. Adv Exp Med Biol. 2012;727:89–98.CrossRefPubMedGoogle Scholar
  23. 23.
    Amsen D, Antov A, Flavell RA. The different faces of Notch in T-helper-cell differentiation. Nat Rev Immunol. 2009;9:116–24.CrossRefPubMedGoogle Scholar
  24. 24.
    Bailis W, Yashiro-Ohtani Y, Fang TC, Hatton RD, Weaver CT, Artis D, et al. Notch simultaneously orchestrates multiple helper T cell programs independently of cytokine signals. Immunity. 2013;39:148–59.CrossRefPubMedCentralPubMedGoogle Scholar
  25. 25.
    Liu T, Hu B, Choi YY, Chung M, Ullenbruch M, Yu H, et al. Notch1 signaling in FIZZ1 induction of myofibroblast differentiation. Am J Pathol. 2009;174:1745–55.CrossRefPubMedCentralPubMedGoogle Scholar
  26. 26.
    Aoyagi-Ikeda K, Maeno T, Matsui H, Ueno M, Hara K, Aoki Y, et al. Notch induces myofibroblast differentiation of alveolar epithelial cells via transforming growth factor-{beta}-Smad3 pathway. Am J Respir Cell Mol Biol. 2011;45:136–44.PubMedGoogle Scholar
  27. 27.
    Schrier DJ, Kunkel RG, Phan SH. The role of strain variation in murine bleomycin-induced pulmonary fibrosis. Am Rev Respir Dis. 1983;127:63–6.PubMedGoogle Scholar
  28. 28.
    Harrison JH Jr, Lazo JS. Plasma and pulmonary pharmacokinetics of bleomycin in murine strains that are sensitive and resistant to bleomycin-induced pulmonary fibrosis. J Pharmacol Exp Ther. 1988;247:1052–8.PubMedGoogle Scholar
  29. 29.
    Yao H, Edirisinghe I, Rajendrasozhan S, Chiba Y, Mizuguchi J, Yoshimoto T. Cigarette smoke-mediated inflammatory and oxidative responses are strain-dependent in mice. Am J Physiol Lung Cell Mol Physiol. 2008;294:L1174–86.CrossRefPubMedGoogle Scholar
  30. 30.
    Moore BB, Hogaboam CM. Murine models of pulmonary fibrosis. Am J Physiol Lung Cell Mol Physiol. 2008;294:L152–60.CrossRefPubMedGoogle Scholar
  31. 31.
    Thurlbeck WM. Measurement of pulmonary emphysema. Am Rev Respir Dis. 1967;95:752–64.PubMedGoogle Scholar
  32. 32.
    Thurlbeck WM. The internal surface area of nonemphysematous lungs. Am Rev Respir Dis. 1967;95:765–73.PubMedGoogle Scholar
  33. 33.
    Ashcroft T, Simpson JM, Timbrell V. Simple method of estimating severity of pulmonary fibrosis on a numerical scale. J Clin Pathol. 1988;41:467–70.CrossRefPubMedCentralPubMedGoogle Scholar
  34. 34.
    Thrall RS, Phan SH, McCormick JR, Ward PA. The development of bleomycin-induced pulmonary fibrosis in neutrophil-depleted and complement-depleted rats. Am J Pathol. 1981;105:76–81.PubMedCentralPubMedGoogle Scholar
  35. 35.
    Clark JG, Kuhn C 3rd. Bleomycin-induced pulmonary fibrosis in hamsters: effect of neutrophil depletion on lung collagen synthesis. Am Rev Respir Dis. 1982;126:737–9.PubMedGoogle Scholar
  36. 36.
    Osanai K, Takahashi K, Suwabe A, Takada K, Ikeda H, Sato S, et al. The effect of cigarette smoke on bleomycin-induced pulmonary fibrosis in hamsters. Am Rev Respir Dis. 1988;138:1276–81.CrossRefPubMedGoogle Scholar
  37. 37.
    Cisneros-Lira J, Gaxiola M, Ramos C, Selman M, Pardo A. Cigarette smoke exposure potentiates bleomycin-induced lung fibrosis in guinea pigs. Am J Physiol Lung Cell Mol Physiol. 2003;285:L949–56.CrossRefPubMedGoogle Scholar
  38. 38.
    Hogg JC, Chu F, Utokaparch S, Woods R, Elliott WM, Buzatu L, et al. The nature of small-airway obstruction in chronic obstructive pulmonary disease. N Engl J Med. 2004;350:2645–53.CrossRefPubMedGoogle Scholar
  39. 39.
    Demoor T, Bracke KR, Vermaelen KY, Dupont L, Joos GF, Brusselle GG. CCR7 modulates pulmonary and lymph node inflammatory responses in cigarette smoke-exposed mice. J Immunol. 2009;183:8186–94.CrossRefPubMedGoogle Scholar
  40. 40.
    Zhu J, Kaplan AM, Goud SN. Immunologic alterations in bleomycin-treated mice: role of pulmonary fibrosis in the modulation of immune responses. Am J Respir Crit Care Med. 1996;153:1924–30.CrossRefPubMedGoogle Scholar
  41. 41.
    Trujillo G, Hartigan AJ, Hogaboam CM. T regulatory cells and attenuated bleomycin-induced fibrosis in lungs of CCR7-/- mice. Fibrogenesis Tissue Repair. 2010;3:18.CrossRefPubMedCentralPubMedGoogle Scholar
  42. 42.
    van der Strate BW, Postma DS, Brandsma CA, Melgert BN, Luinge MA, Geerlings M, et al. Cigarette smoke-induced emphysema: a role for the B cell. Am J Respir Crit Care Med. 2006;173:751–8.CrossRefPubMedGoogle Scholar
  43. 43.
    Maekawa Y, Tsukumo S, Chiba S, Hirai H, Hayashi Y, Okada H, et al. Delta1-Notch3 interactions bias the functional differentiation of activated CD4+ T cells. Immunity. 2003;19:549–59.CrossRefPubMedGoogle Scholar
  44. 44.
    Krawczyk CM, Sun J, Pearce EJ. Th2 differentiation is unaffected by Jagged2 expression on dendritic cells. J Immunol. 2008;180:7931–7.CrossRefPubMedCentralPubMedGoogle Scholar
  45. 45.
    Sun J, Krawczyk CJ, Pearce EJ. Suppression of Th2 cell development by Notch ligands Delta1 and Delta4. J Immunol. 2008;180:1655–61.CrossRefPubMedGoogle Scholar
  46. 46.
    Jiang D, Liang J, Hodge J, Lu B, Zhu Z, Yu S, et al. Regulation of pulmonary fibrosis by chemokine receptor CXCR3. J Clin Invest. 2004;114:291–9.CrossRefPubMedCentralPubMedGoogle Scholar

Copyright information

© Springer Basel 2015

Authors and Affiliations

  • Shi Li
    • 1
  • Xiaofei Hu
    • 2
  • Zheng Wang
    • 3
  • Meng Wu
    • 1
  • Jinnong Zhang
    • 1
    Email author
  1. 1.Department of Respiratory Medicine, Union Hospital, Tongji Medical CollegeHuazhong University of Science and TechnologyWuhanChina
  2. 2.Department of Respiratory MedicineXiaogan Central HospitalXiaoganChina
  3. 3.Department of Respiratory MedicinePeople’s Hospital of Henan ProvinceZhengzhouChina

Personalised recommendations