Azithromycin Inhibits MUC5AC Production Induced by the Pseudomonas aeruginosa Autoinducer N-(3-Oxododecanoyl) Homoserine Lactone in NCI-H292 Cells

ABSTRACT The features of chronic airway diseases, including chronic bronchitis, cystic fibrosis, bronchiectasis, and diffuse panbronchiolitis, include chronic bacterial infection and airway obstruction by mucus. Pseudomonas aeruginosa is one of the most common pathogens in chronic lung infection, and quorum-sensing systems contribute to the pathogenesis of this disease. The quorum-sensing signal molecule [N-(3-oxododecanoyl) homoserine lactone (3O-C12-HSL)] not only regulates bacterial virulence but also is associated with the immune response. In this study, we investigated whether 3O-C12-HSL could stimulate the production of a major mucin core protein, MUC5AC. The effect of a macrolide on MUC5AC production was also studied. 3O-C12-HSL induced NCI-H292 cells to express MUC5AC at both the mRNA and the protein levels in time- and dose-dependent manners. A 15-membered macrolide, azithromycin, inhibited MUC5AC production that was activated by 3O-C12-HSL. 3O-C12-HSL induced extracellular signal-regulated kinase (ERK) 1/2 and I-κB phosphorylation in cells, and this induction was suppressed by azithromycin. 3O-C12-HSL-induced MUC5AC production was blocked by the ERK pathway inhibitor PD98059. Our findings suggest that the P. aeruginosa autoinducer 3O-C12-HSL contributes to excessive mucin production in chronic bacterial infection. Azithromycin seems to reduce this mucin production by interfering with intracellular signal transduction.

[1]  J. Davies,et al.  MUC5AC, but not MUC2, is a prominent mucin in respiratory secretions , 1996, Glycoconjugate Journal.

[2]  Y. Hirakata,et al.  Clarithromycin inhibits overproduction of muc5ac core protein in murine model of diffuse panbronchiolitis. , 2003, American journal of physiology. Lung cellular and molecular physiology.

[3]  L. Saiman,et al.  Azithromycin in patients with cystic fibrosis chronically infected with Pseudomonas aeruginosa: a randomized controlled trial. , 2003, JAMA.

[4]  Si-Youn Song,et al.  Interleukin-1beta induces MUC2 and MUC5AC synthesis through cyclooxygenase-2 in NCI-H292 cells. , 2002, Molecular pharmacology.

[5]  Roger S Smith,et al.  The Pseudomonas Autoinducer N-(3-Oxododecanoyl) Homoserine Lactone Induces Cyclooxygenase-2 and Prostaglandin E2 Production in Human Lung Fibroblasts: Implications for Inflammation1 , 2002, The Journal of Immunology.

[6]  T. Nukiwa,et al.  Clarithromycin suppresses lipopolysaccharide-induced interleukin-8 production by human monocytes through AP-1 and NF-kappa B transcription factors. , 2002, The Journal of antimicrobial chemotherapy.

[7]  Richard Phipps,et al.  The Pseudomonas aeruginosa Quorum-Sensing Molecule N-(3-Oxododecanoyl)Homoserine Lactone Contributes to Virulence and Induces Inflammation In Vivo , 2002, Journal of bacteriology.

[8]  T. A. Springer,et al.  IL-8 Production in Human Lung Fibroblasts and Epithelial Cells Activated by the Pseudomonas Autoinducer N-3-Oxododecanoyl Homoserine Lactone Is Transcriptionally Regulated by NF-κB and Activator Protein-21 , 2001, The Journal of Immunology.

[9]  K. Dabbagh,et al.  IL-13 induces mucin production by stimulating epidermal growth factor receptors and by activating neutrophils. , 2001, American journal of physiology. Lung cellular and molecular physiology.

[10]  K. Yanagihara,et al.  Lipopolysaccharide Induces Mucus Cell Metaplasia in Mouse Lung. , 2001, American journal of respiratory cell and molecular biology.

[11]  Matthew R. Parsek,et al.  Quorum-sensing signals indicate that cystic fibrosis lungs are infected with bacterial biofilms , 2000, Nature.

[12]  S. Kjelleberg,et al.  A novel and sensitive method for the quantification of N-3-oxoacyl homoserine lactones using gas chromatography-mass spectrometry: application to a model bacterial biofilm. , 2000, Environmental microbiology.

[13]  B. Iglewski,et al.  Bacterial Quorum Sensing in Pathogenic Relationships , 2000, Infection and Immunity.

[14]  K. Dabbagh,et al.  Oxidative Stress Causes Mucin Synthesis Via Transactivation of Epidermal Growth Factor Receptor: Role of Neutrophils1 , 2000, The Journal of Immunology.

[15]  J. Davies,et al.  Identification of MUC5B, MUC5AC and small amounts of MUC2 mucins in cystic fibrosis airway secretions. , 1999, The Biochemical journal.

[16]  S. Wenzel,et al.  Allergen-induced IL-9 directly stimulates mucin transcription in respiratory epithelial cells. , 1999, The Journal of clinical investigation.

[17]  K. Dabbagh,et al.  IL-4 induces mucin gene expression and goblet cell metaplasia in vitro and in vivo. , 1999, Journal of immunology.

[18]  C. Agustí,et al.  Epidermal growth factor system regulates mucin production in airways. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[19]  J. A. Linton,et al.  Effects of TNF-alpha and IL-1 beta on mucin, lysozyme, IL-6 and IL-8 in passage-2 normal human nasal epithelial cells. , 1999, Acta oto-laryngologica.

[20]  S. Kudoh,et al.  Improvement of survival in patients with diffuse panbronchiolitis treated with low-dose erythromycin. , 1998, American journal of respiratory and critical care medicine.

[21]  Jian-Dong Li,et al.  Activation of NF-kappaB via a Src-dependent Ras-MAPK-pp90rsk pathway is required for Pseudomonas aeruginosa-induced mucin overproduction in epithelial cells. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[22]  Jian-Dong Li,et al.  Mucin gene (MUC 2 and MUC 5AC) upregulation by Gram-positive and Gram-negative bacteria. , 1998, Biochimica et biophysica acta.

[23]  D. Wheeler,et al.  The Pseudomonas aeruginosaQuorum-Sensing Signal MoleculeN-(3-Oxododecanoyl)-l-Homoserine Lactone Has Immunomodulatory Activity , 1998, Infection and Immunity.

[24]  R. Flavell,et al.  A novel role for murine IL-4 in vivo: induction of MUC5AC gene expression and mucin hypersecretion. , 1997, American journal of respiratory cell and molecular biology.

[25]  J. D. Li,et al.  Transcriptional activation of mucin by Pseudomonas aeruginosa lipopolysaccharide in the pathogenesis of cystic fibrosis lung disease. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[26]  M. Kondo,et al.  Lipopolysaccharide-induced goblet cell hypersecretion in the guinea pig trachea: inhibition by macrolides. , 1997, The American journal of physiology.

[27]  G. Campbell,et al.  Intrapulmonary pharmacokinetics of azithromycin in healthy volunteers given five oral doses , 1996, Antimicrobial agents and chemotherapy.

[28]  A. Prince,et al.  Diverse Pseudomonas aeruginosa gene products stimulate respiratory epithelial cells to produce interleukin-8. , 1995, The Journal of clinical investigation.

[29]  J. Reiser,et al.  Autoinducer-mediated regulation of rhamnolipid biosurfactant synthesis in Pseudomonas aeruginosa. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[30]  E. Greenberg,et al.  A second N-acylhomoserine lactone signal produced by Pseudomonas aeruginosa. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[31]  S. Gendler,et al.  Epithelial mucin genes. , 1995, Annual review of physiology.

[32]  E. Greenberg,et al.  Structure of the autoinducer required for expression of Pseudomonas aeruginosa virulence genes. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[33]  M. Gambello,et al.  LasR of Pseudomonas aeruginosa is a transcriptional activator of the alkaline protease gene (apr) and an enhancer of exotoxin A expression , 1993, Infection and immunity.

[34]  C. Basbaum,et al.  Mucin in disease. Modification of mucin gene expression in airway disease. , 1991, The American review of respiratory disease.

[35]  S. Goswami,et al.  Erythromycin inhibits respiratory glycoconjugate secretion from human airways in vitro. , 1990, The American review of respiratory disease.

[36]  J. Shelhamer,et al.  Human respiratory mucus. , 1984, The American review of respiratory disease.