title: Acetylcholine mediates the release of IL-8 in human bronchial epithelial cells by a NFkB/ERK-dependent mechanism.
abstract: Acetylcholine may play a role in cell activation and airway inflammation. We evaluated the levels of both mRNA and protein of muscarinic M(1), M(2), M(3) receptors in human bronchial epithelial cell line (16HBE). 16HBE cells were also stimulated with acetylcholine and extracellular signal-regulated kinase1/2 (ERK1/2) and NFkB pathway activation as well as the IL-8 release was assessed in the presence or absence of the inhibitor of Protein-kinase (PKC) (GF109203X), of the inhibitor of mitogenic activated protein-kinase kinase (MAPKK) (PDO9805), of the inhibitor of kinaseB-alpha phosphorilation (pIkBalpha) (BAY11-7082), and of muscarinic receptor antagonists tiotropium bromide, 4-Diphenylacetoxy-N-methylpiperidine methiodide (4-DAMP), telenzepine, gallamine. Additionally, we tested the IL-8-mediated neutrophil chemotactic activity of 16HBE supernatants stimulated with acetylcholine in the presence or absence of tiotropium. 16HBE cells expressed both protein and mRNA for muscarinic M(3), M(2) and M(1) receptors with levels of muscarinic M(3) receptor>muscarinic M(1) receptor>muscarinic M(2) receptor. Acetylcholine (10 microM) significantly stimulated ERK1/2 and NFkB activation as well as IL-8 release in 16HBE cells when compared to basal values. Furthermore, while the use of tiotropium, 4-DAMP, GF109203X, PDO98059, BAY11-7082 completely abolished these events, the use of telenzepine and gallamine were only partially able to downregulate these effects. Additionally, acetylcholine-mediated IL-8 release from 16HBE cells significantly increased chemotaxis toward neutrophils and this effect was blocked by tiotropium. In conclusion, acetylcholine activates the release of IL-8 from 16HBE involving PKC, ERK1/2 and NFkB pathways via muscarinic receptors, suggesting that it is likely to contribute to IL-8 related neutrophilic inflammatory disorders in the airway. Thus, muscarinic antagonists may contribute to control inflammatory processes in airway diseases.
authors.affiliation: Institute of Biomedicine and Molecular Immunology, Italian National Research Council, Palermo, Italy. profita@ibim.cnr.it
authors.forename: Mirella
authors.initials: M
authors.lastname: Profita
authors.forename: Anna
authors.initials: A
authors.lastname: Bonanno
authors.forename: Liboria
authors.initials: L
authors.lastname: Siena
authors.forename: Maria
authors.initials: M
authors.lastname: Ferraro
authors.forename: Angela M
authors.initials: AM
authors.lastname: Montalbano
authors.forename: Flora
authors.initials: F
authors.lastname: Pompeo
authors.forename: Loredana
authors.initials: L
authors.lastname: Riccobono
authors.forename: Michael P
authors.initials: MP
authors.lastname: Pieper
authors.forename: Mark
authors.initials: M
authors.lastname: Gjomarkaj
mesh_terms: D000109:Acetylcholine / Q000494:pharmacology* / Q000502:physiology; D001980:Bronchi / Q000166:cytology; D002461:Cell Line, Transformed; D002634:Chemotaxis, Leukocyte; D004847:Epithelial Cells / Q000187:drug effects* / Q000378:metabolism; D005419:Flavonoids / Q000494:pharmacology; D005703:Gallamine Triethiodide / Q000494:pharmacology; D006801:Humans; D007211:Indoles / Q000494:pharmacology; D016209:Interleukin-8 / Q000378:metabolism*; D008301:Maleimides / Q000494:pharmacology; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism*; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism*; D018727:Muscarinic Antagonists / Q000494:pharmacology; D016328:NF-kappa B / Q000378:metabolism*; D009504:Neutrophils / Q000187:drug effects / Q000502:physiology; D009570:Nitriles / Q000494:pharmacology; D010880:Piperidines / Q000494:pharmacology; D010890:Pirenzepine / Q000031:analogs & derivatives / Q000494:pharmacology; D011493:Protein Kinase C / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D011976:Receptors, Muscarinic / Q000235:genetics / Q000378:metabolism; D012602:Scopolamine Derivatives / Q000494:pharmacology; D013450:Sulfones / Q000494:pharmacology; D000069447:Tiotropium Bromide
chemical_list: C434003:3-(4-methylphenylsulfonyl)-2-propenenitrile; D005419:Flavonoids; D007211:Indoles; D016209:Interleukin-8; D008301:Maleimides; D018727:Muscarinic Antagonists; D016328:NF-kappa B; D009570:Nitriles; D010880:Piperidines; D012333:RNA, Messenger; D011976:Receptors, Muscarinic; D012602:Scopolamine Derivatives; D013450:Sulfones; C046338:telenzepine; D010890:Pirenzepine; C042375:4-diphenylacetoxy-1,1-dimethylpiperidinium; D011493:Protein Kinase C; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; C070515:bisindolylmaleimide I; D000109:Acetylcholine; D005703:Gallamine Triethiodide; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one; D000069447:Tiotropium Bromide