title: Bradykinin- and lipopolysaccharide-induced bradykinin B2 receptor expression, interleukin 8 release and "nitrosative stress" in bronchial epithelial cells BEAS-2B: role for neutrophils.
abstract: Bradykinin-induced interleukin (IL)-8 release should potentially activate neutrophils releasing myeloperoxidase (MPO) and subsequently generating "nitrosative stress". We studied bradykinin-induced expression of bradykinin B(2) receptor and bradykinin- and lipopolysaccharide (LPS)-induced IL-8 release, MPO (marker of neutrophil activation) and 3-nitrotyrosine (3-NT; marker of "nitrosative stress") production in human bronchial epithelial cells BEAS-2B alone or in co-culture with human neutrophils. We evaluated B(2) receptor protein expression in BEAS-2B cells by immunostainings and Western blot analysis, and measured respectively bradykinin- or LPS-induced IL-8 release in BEAS-2B cells and bradykinin- and/or LPS-induced MPO and 3-NT production in BEAS-2B cells co-cultured with human neutrophils by ELISA. In addition, we evaluated bradykinin- and/or LPS-induced 3-NT formation in BEAS-2B cells co-cultured with human neutrophils by immunocytochemistry. Bradykinin up-regulates B(2) receptor expression (P<0.05) and stimulate IL-8 release (P<0.001) in BEAS-2B cells. Either the selective bradykinin B(2) receptor antagonist HOE 140 or the selective bradykinin B(1) receptor antagonist Lys-(des-Arg(9), Leu(8))-bradykinin alone halved IL-8 release and the combination of both drugs suppressed this effect. In BEAS-2B cells co-cultured with human neutrophils bradykinin increased MPO release and 3-NT production compared to BEAS-2B cells with human neutrophils (P<0.001), and the addition of LPS in BEAS-2B cells with human neutrophils and bradykinin induced a further dramatically increase of MPO release and 3-NT formation (P<0.001). Bradykinin and LPS provoked "nitrosative stress", potentially mediated by IL-8, in bronchial epithelium co-cultured with neutrophils suggesting a role for bradykinin in the amplification of chronic airway inflammation via production of "nitrosative stress".
authors.affiliation: Department of Clinical and Biological Sciences, Division of Respiratory Disease, University of Torino, Torino, Italy. fabioluigimassimo.ricciardolo@unito.it
authors.forename: Fabio L M
authors.initials: FL
authors.lastname: Ricciardolo
authors.forename: Valentina
authors.initials: V
authors.lastname: Sorbello
authors.forename: Sabrina
authors.initials: S
authors.lastname: Benedetto
authors.forename: Ilaria
authors.initials: I
authors.lastname: Defilippi
authors.forename: Federica
authors.initials: F
authors.lastname: Sabatini
authors.forename: Andrea
authors.initials: A
authors.lastname: Robotti
authors.forename: Dein C
authors.initials: DC
authors.lastname: van Renswouw
authors.forename: Caterina
authors.initials: C
authors.lastname: Bucca
authors.forename: Gert
authors.initials: G
authors.lastname: Folkerts
authors.forename: Virginia
authors.initials: V
authors.lastname: De Rose
mesh_terms: D001920:Bradykinin / Q000494:pharmacology*; D001980:Bronchi / Q000166:cytology; D002460:Cell Line; D018920:Coculture Techniques; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D016209:Interleukin-8 / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000494:pharmacology*; D009504:Neutrophils / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D009195:Peroxidase / Q000378:metabolism; D026361:Reactive Nitrogen Species / Q000378:metabolism*; D043782:Receptor, Bradykinin B2 / Q000378:metabolism*; D014443:Tyrosine / Q000031:analogs & derivatives / Q000096:biosynthesis / Q000378:metabolism
chemical_list: D016209:Interleukin-8; D008070:Lipopolysaccharides; D026361:Reactive Nitrogen Species; D043782:Receptor, Bradykinin B2; C002744:3-nitrotyrosine; D014443:Tyrosine; D009195:Peroxidase; D001920:Bradykinin