Archives of Pharmacal Research

, Volume 37, Issue 4, pp 520–529 | Cite as

Ambroxol inhibits rhinovirus infection in primary cultures of human tracheal epithelial cells

  • Mutsuo Yamaya
  • Hidekazu Nishimura
  • Lusamba Kalonji Nadine
  • Chiharu Ota
  • Hiroshi Kubo
  • Ryoichi Nagatomi
Research Article

Abstract

The mucolytic drug ambroxol hydrochloride reduces the production of pro-inflammatory cytokines and the frequency of exacerbation in patients with chronic obstructive pulmonary disease (COPD). However, the inhibitory effects of ambroxol on rhinovirus infection, the major cause of COPD exacerbations, have not been studied. We examined the effects of ambroxol on type 14 rhinovirus (RV14) infection, a major RV group, in primary cultures of human tracheal epithelial cells. RV14 infection increased virus titers and cytokine content in the supernatants and RV14 RNA in the cells. Ambroxol (100 nM) reduced RV14 titers and cytokine concentrations of interleukin (IL)-1β, IL-6 and IL-8 in the supernatants and RV14 RNA in the cells after RV14 infection, in addition to reducing susceptibility to RV14 infection. Ambroxol also reduced the expression of intercellular adhesion molecule-1 (ICAM-1), the receptor for RV14, and the number of acidic endosomes from which RV14 RNA enters the cytoplasm. In addition, ambroxol reduced the activation of the transcription factor nuclear factor kappa B (NF-κB) in the nucleus. These results suggest that ambroxol inhibits RV14 infection partly by reducing ICAM-1 and acidic endosomes via the inhibition of NF-κB activation. Ambroxol may modulate airway inflammation by reducing the production of cytokines in rhinovirus infection.

Keywords

Airway epithelium Ambroxol Infection control Inflammation Intercellular adhesion molecule-1 (ICAM-1) Rhinovirus 

References

  1. Akira, S., T. Hirano, T. Taga, and T. Kishimoto. 1990. Biology of multifunctional cytokines: IL 6 and related molecules (IL 1 and TNF). The FASEB Journal 4: 2860–2867.Google Scholar
  2. Blesa, S., J. Cortijo, M. Mata, A. Serrano, D. Closa, F. Santangelo, J.M. Estrela, J. Suchankova, and E.J. Morcillo. 2003. Oral N-acetylcysteine attenuates the rat pulmonary inflammatory response to antigen. European Respiratory Journal 21: 394–400.PubMedCrossRefGoogle Scholar
  3. Casasnovas, J.M., and T.A. Springer. 1994. Pathway of rhinovirus disruption by soluble intercellular adhesion molecule 1 (ICAM-1): An intermediate in which ICAM-1 is bound and RNA is released. Journal of Virology 68: 5882–5889.PubMedCentralPubMedGoogle Scholar
  4. Condit, R.C. 2006. Principles of virology. In Fields virology, 5th ed, ed. D.M. Knipe, and P.M. Howley, 25–57. Philadelphia: Lippincott Williams and Wilkins.Google Scholar
  5. Fernandez, R., J.R. Bosqueiro, A.C. Cassola, and G. Malnic. 1997. Role of Cl in electrogenic H+ secretion by cortical distal tubule. Journal of Membrane Biology 157: 193–201.PubMedCrossRefGoogle Scholar
  6. Gibbs, B.F., W. Schmutzler, I.B. Vollrath, P. Brosthardt, U. Braam, H.H. Wolff, and G. Zwadlo-Klarwasser. 1999. Ambroxol inhibits the release of histamine, leukotrienes and cytokines from human leukocytes and mast cells. Inflammation Research 48: 86–93.PubMedCrossRefGoogle Scholar
  7. Greve, J.M., G. Davis, A.M. Meyer, C.P. Forte, S.C. Yost, C.W. Marlor, M.E. Kamarck, and A. McClelland. 1989. The major human rhinovirus receptor is ICAM-1. Cell 56: 839–847.PubMedCrossRefGoogle Scholar
  8. Hasegawa, I., N. Niisato, Y. Iwasaki, and Y. Marunaka. 2006. Ambroxol-induced modification of ion transport in human airway Calu-3 epithelia. Biochemical and Biophysical Research Communications 343: 475–482.PubMedCrossRefGoogle Scholar
  9. Hong, J.S., H.H. Ko, E.S. Han, and C.S. Lee. 2003. Inhibition of bleomycin-induced cell death in rat alveolar macrophages and human lung epithelial cells by ambroxol. Biochemical Pharmacology 66: 1297–1306.PubMedCrossRefGoogle Scholar
  10. Jang, Y.Y., J.H. Song, Y.K. Shin, E.S. Han, and C.S. Lee. 2003. Depressant effects of ambroxol and erdosteine on cytokine synthesis, granule enzyme release, and free radical production in rat alveolar macrophages activated by lipopolysaccharide. Pharmacology and Toxicology 92: 173–179.PubMedCrossRefGoogle Scholar
  11. Malerba, M., A. Ponticiello, A. Radaeli, G. Bensi, and V. Grassi. 2004. Effect of twelve-months therapy with oral ambroxol in preventing exacerbations in patients with COPD. Double-blind, randomized, multicenter, placebo-controlled study (the AMETHIST Trial). Pulmonary Pharmacology & Therapeutics 17: 27–34.CrossRefGoogle Scholar
  12. Marshansky, V., and P. Vinay. 1996. Proton gradient formation in early endosomes from proximal tubes. Biochimica et Biophysica Acta 1284: 171–180.PubMedCrossRefGoogle Scholar
  13. Mellman, I., R. Fuchs, and A. Helenius. 1986. Acidification of the endocytic and exocytic pathways. Annual Review of Biochemistry 55: 663–700.PubMedCrossRefGoogle Scholar
  14. Mezzetti, M., L. Colombo, M.G. Marini, V. Crusi, P. Pierfederici, and E. Mussini. 1990. A pharmacokinetic study on pulmonary tropism of ambroxol in patients under thoracic surgery. The European Journal of Emergency Surgery and Intensive Care 13: 179–185.Google Scholar
  15. Moulin, P., T. Igarashi, P. Van der Smissen, J.P. Cosyns, P. Verroust, R.V. Thakker, S.J. Scheinman, P.J. Courtoy, and O. Devuyst. 2003. Altered polarity and expression of H+-ATPase without ultrastructural changes in kidneys of Dent’s disease patients. Kidney International 63: 1285–1295.PubMedCrossRefGoogle Scholar
  16. Nass, R., and R. Rao. 1998. Novel localization of a Na+/H+ exchanger in a late endosomal compartment of yeast. Implications for vacuole biogenesis. Journal of Biological Chemistry 273: 21054–21060.PubMedCrossRefGoogle Scholar
  17. Natajaran, K., S. Singh, T.R. Burke Jr, D. Grunberger, and B.B. Aggarwal. 1996. Caffeic acid phenethyl ester is a potent and specific inhibitor of activation of nuclear transcription factor NF-kappa B. Proceedings of the National Academy of Sciences of the United States of America 93: 9090–9095.CrossRefGoogle Scholar
  18. Nobata, K., M. Fujimura, Y. Ishiura, S. Myou, and S. Nakao. 2006. Ambroxol for the prevention of acute upper respiratory disease. Clinical and Experimental Medicine 6: 79–83.PubMedCrossRefGoogle Scholar
  19. Nolan, T., R.E. Hands, and S.A. Bustin. 2006. Quantification of mRNA using real-time RT-PCR. Nature Protocols 1: 1559–1582.PubMedCrossRefGoogle Scholar
  20. Numazaki, Y., T. Oshima, A. Ohmi, A. Tanaka, Y. Oizumi, S. Komatsu, T. Takagi, M. Karahashi, and N. Ishida. 1987. A microplate methods for isolation of viruses from infants and children with acute respiratory infections. Microbiology and Immunology 31: 1085–1095.PubMedCrossRefGoogle Scholar
  21. Olivieri, D., G. Zavatti, G. Tomasini, S. Daniotti, G. Bonsignore, G. Ferrara, N. Carnimero, R. Chianese, E. Catena, S. Marcatili, M. Del Donno, C. Grassi, E. Pozzi, V. Grassi, C. Tantucci, M. Lucchesi, G. Schimid, C.F. Marchioni, S. Penitenti, A. Mistretta, N. Crimi, L. Casali, R. Cabiddu, C. Donner, A. Patessio, V. Massei, C.M. Sanguinetti, O. Orlandi, S. Bruna, C. Serra, and A. Giacopelli. 1987. Ambroxol for the prevention of chronic bronchitis: Exacerbations: Long-term multicenter trial. Respiration 51: 42–51.PubMedCrossRefGoogle Scholar
  22. Papi, A., and S.L. Johnston. 1999. Respiratory epithelial cell expression of vascular cell adhesion molecule-1 and its up-regulation by rhinovirus infection via NF-κB and GATA transcription factors. Journal of Biological Chemistry 274: 30041–30051.PubMedCrossRefGoogle Scholar
  23. Pérez, L., and L. Carrasco. 1993. Entry of poliovirus into cells does not require a low-pH step. Journal of Virology 67: 4543–4548.PubMedCentralPubMedGoogle Scholar
  24. Seemungal, T., R. Harper-Owen, A. Bhowmik, D.J. Jeffries, and J.A. Wedzicha. 2000. Detection of rhinovirus in induced sputum at exacerbation of chronic obstructive pulmonary disease. European Respiratory Journal 16: 677–683.PubMedCrossRefGoogle Scholar
  25. Seemungal, T., R. Harper-Owen, A. Bhowmik, I. Moric, G. Sanderson, S. Message, P. Maccallum, T.W. Meade, D.J. Jeffries, S.L. Johnston, and J.A. Wedzicha. 2001. Respiratory viruses, symptoms, and inflammatory markers in acute exacerbations and stable chronic obstructive pulmonary disease. American Journal of Respiratory and Critical Care Medicine 164: 1618–1623.PubMedCrossRefGoogle Scholar
  26. Sethi, S. 2004. New developments in the pathogenesis of acute exacerbations of chronic obstructive pulmonary disease. Current Opinion in Infectious Diseases 17: 113–119.PubMedCrossRefGoogle Scholar
  27. Severina, I.S., O.G. Bussygina, N.V. Pyatakova, Y.V. Khropov, and R.A. Krasnoperov. 2000. Ambroxol as an inhibitor of nitric oxide-dependent activation of soluble guanylate cyclase. European Journal of Pharmacology 407: 61–64.PubMedCrossRefGoogle Scholar
  28. Su, F., F. Wang, W. Gao, and H. Li. 2007. Determination of ambroxol in human plasma by high performance liquid chromatography-electrospray ionization mass spectrometry (HPLC-MS/ESI). Journal of Chromatography B 853: 364–368.CrossRefGoogle Scholar
  29. Su, X., L. Wang, Y. Song, and C. Bai. 2004. Inhibition of inflammatory responses by ambroxol, a mucolytic agent, in a murine model of acute lung injury induced by lipopolysaccharide. Intensive Care Medicine 30: 133–140.PubMedCrossRefGoogle Scholar
  30. Subauste, M.C., D.B. Jacoby, S.M. Richards, and D. Proud. 1995. Infection of a human respiratory epithelial cell line with rhinovirus. Induction of cytokine release and modulation of susceptibility to infection by cytokine exposure. Journal of Clinical Investigation 96: 549–557.PubMedCentralPubMedCrossRefGoogle Scholar
  31. Suzuki, T., M. Yamaya, K. Sekizawa, M. Hosoda, N. Yamada, S. Ishizuka, K. Nakayama, M. Yanai, Y. Numazaki, and H. Sasaki. 2001. Bafilomycin A1 inhibits rhinovirus infection in human airway epithelium: effects on endosome and ICAM-1. American Journal of Physiology 280: L1115–L1127.PubMedGoogle Scholar
  32. Suzuki, T., M. Yamaya, K. Sekizawa, M. Hosoda, N. Yamada, S. Ishizuka, A. Yoshino, H. Yasuda, H. Takahashi, H. Nishimura, and H. Sasaki. 2002. Erythromycin inhibits rhinovirus infection in cultured human tracheal epithelial cells. American Journal of Respiratory and Critical Care Medicine 165: 1113–1118.PubMedCrossRefGoogle Scholar
  33. Suzuki, T., M. Yamaya, K. Sekizawa, N. Yamada, K. Nakayama, S. Ishizuka, M. Kamanaka, T. Morimoto, Y. Numazaki, and H. Sasaki. 2000. Effects of dexamethasone on rhinovirus infection in cultured human tracheal epithelial cells. American Journal of Physiology 278: L560–L571.PubMedGoogle Scholar
  34. Teramoto, S., M. Suzuki, E. Ohga, H. Ishii, T. Matsuse, and Y. Ouchi. 1999. Effects of ambroxol on spontaneous or stimulated generation of reactive oxygen species by bronchoalveolar lavage cells harvested from patients with or without chronic obstructive pulmonary diseases. Pharmacology 59: 135–141.PubMedCrossRefGoogle Scholar
  35. Turner, R.B., and R.B. Couch. 2006. Rhinoviruses. In Fields Virology, 5th ed, ed. D.M. Knipe, and P.M. Howley, 895–909. Philadelphia: Lippincott Williams and Wilkins.Google Scholar
  36. van de Stolpe, A., and P.T. van der Saag. 1996. Intercellular adhesion molecule-1. Journal of Molecular Medicine 74: 13–33.PubMedCrossRefGoogle Scholar
  37. Yamaya, M., H. Nishimura, Y. Hatachi, H. Yasuda, X. Deng, T. Sasaki, H. Kubo, and R. Nagatomi. 2012. Inhibitory effects of tiotropium on rhinovirus infection in human airway epithelial cells. European Respiratory Journal 40: 122–132.PubMedCrossRefGoogle Scholar
  38. Yamaya, M., H. Nishimura, Y. Hatachi, M. Yoshida, H. Fujiwara, M. Asada, K. Nakayama, H. Yasuda, X. Deng, T. Sasaki, H. Kubo, and R. Nagatomi. 2011. Procaterol inhibits rhinovirus infection in primary cultures of human tracheal epithelial cells. European Journal of Pharmacology 650: 431–444.PubMedCrossRefGoogle Scholar
  39. Yang, B., D.F. Yao, M. Ohuchi, M. Ide, M. Yano, Y. Okumura, and H. Kido. 2002. Ambroxol suppresses influenza-virus proliferation in the mouse airway by increasing antiviral factor levels. European Respiratory Journal 19: 952–958.PubMedCrossRefGoogle Scholar
  40. Yasuda, H., M. Yamaya, T. Sasaki, D. Inoue, K. Nakayama, M. Yamada, M. Asada, M. Yoshida, T. Suzuki, H. Nishimura, and H. Sasaki. 2006. Carbocisteine inhibits rhinovirus infection in human tracheal epithelial cells. European Respiratory Journal 28: 51–58.PubMedCrossRefGoogle Scholar
  41. Zhu, Z., W. Tang, A. Ray, Y. Wu, O. Einarsson, M.L. Landry, J. Gwaltney Jr, and J.A. Elias. 1996. Rhinovirus stimulation of interleukin-6 in vivo and in vitro. Evidence for nuclear factor κB-dependent transcriptional activation. Journal of Clinical Investigation 97: 421–430.PubMedCentralPubMedCrossRefGoogle Scholar

Copyright information

© The Pharmaceutical Society of Korea 2013

Authors and Affiliations

  • Mutsuo Yamaya
    • 1
  • Hidekazu Nishimura
    • 2
  • Lusamba Kalonji Nadine
    • 1
  • Chiharu Ota
    • 1
  • Hiroshi Kubo
    • 1
  • Ryoichi Nagatomi
    • 3
  1. 1.Department of Advanced Preventive Medicine for Infectious DiseaseTohoku University Graduate School of MedicineSendaiJapan
  2. 2.Virus Research Center, Clinical Research DivisionSendai National HospitalSendaiJapan
  3. 3.Medicine and Science in Sports and ExerciseTohoku University Graduate School of MedicineSendaiJapan

Personalised recommendations