title: Detection of nitric oxide release induced by bradykinin in guinea pig trachea and main bronchi using a porphyrinic microsensor.
abstract: Indirect evidence using nitric oxide (NO) synthase (NOS) inhibitors suggests that in guinea-pig airways bradykinin releases bronchoprotective NO. In this study, using a recently developed electrochemical method of NO measurement based on a porphyrinic microsensor, we investigated whether bradykinin releases NO from guinea-pig airways and whether the epithelium is the main source of NO. Further, the Ca(2+)-dependence of bradykinin-induced NO release was assessed stimulating airway preparations with bradykinin in Ca(2+)-free conditions. We also studied the immunohistochemical distribution of the Ca(2+)- dependent constitutive isoforms of NOS (constitutive NOS [cNOS]: neuronal and endothelial [ecNOS]) in our preparations. The porphyrinic microsensor was placed in the bathing fluid onto the mucosal surface of tracheal or main bronchial segments. Addition of bradykinin vehicle (0.9% saline) did not cause any detectable change of the baseline signal. Addition of bradykinin caused an upward shift of the baseline that reached a maximum within 1 to 2 s. The amplitude of the response to bradykinin was concentration-dependent between the range 1 nM to 10 microM, with a maximum effect at 10 microM. Bradykinin-induced NO release was higher in tracheal than in main bronchial segments. The selective bradykinin B(2) receptor antagonist D-Arg(0)-[Hyp(3), Thi(5), D-Tic(7), Oic(8)]bradykinin (1 microM) inhibited NO release induced by a submaximum concentration of bradykinin (1 microM). The ability of bradykinin to release NO was markedly reduced in epithelium-denuded segments, and abolished in Ca(2+)-free conditions and after pretreatment with N(G)-monomethyl-L-arginine (100 microM), but not with N(G)-monomethyl-D-arginine. Both cNOS isoforms were present in trachea and main bronchi, ecNOS being the predominant isoform in the epithelium. The study shows that bradykinin via B(2) receptor activation caused a rapid and Ca(2+)-dependent release of NO, mainly, but not exclusively, derived from the epithelium. It also shows that both cNOS isoforms may be involved in bradykinin-evoked NO release.
authors.affiliation: Department of Pulmonology, Leiden University Medical Center, Leiden, The Netherlands. fricciardo@pulmonology.azl.nl
authors.forename: F L
authors.initials: FL
authors.lastname: Ricciardolo
authors.forename: L
authors.initials: L
authors.lastname: Vergnani
authors.forename: S
authors.initials: S
authors.lastname: Wiegand
authors.forename: F
authors.initials: F
authors.lastname: Ricci
authors.forename: N
authors.initials: N
authors.lastname: Manzoli
authors.forename: A
authors.initials: A
authors.lastname: Fischer
authors.forename: S
authors.initials: S
authors.lastname: Amadesi
authors.forename: R
authors.initials: R
authors.lastname: Fellin
authors.forename: P
authors.initials: P
authors.lastname: Geppetti
mesh_terms: D000818:Animals; D015374:Biosensing Techniques; D001920:Bradykinin / Q000031:analogs & derivatives / Q000378:metabolism* / Q000494:pharmacology; D065168:Bradykinin Receptor Antagonists; D001980:Bronchi / Q000378:metabolism*; D004563:Electrochemistry; D006168:Guinea Pigs; D007150:Immunohistochemistry; D007527:Isoenzymes / Q000032:analysis; D008297:Male; D009569:Nitric Oxide / Q000032:analysis / Q000378:metabolism*; D019001:Nitric Oxide Synthase / Q000032:analysis; D052248:Nitric Oxide Synthase Type I; D052250:Nitric Oxide Synthase Type III; D011166:Porphyrins; D043782:Receptor, Bradykinin B2; D014132:Trachea / Q000378:metabolism*
chemical_list: D065168:Bradykinin Receptor Antagonists; D007527:Isoenzymes; D011166:Porphyrins; D043782:Receptor, Bradykinin B2; D009569:Nitric Oxide; C065679:icatibant; D019001:Nitric Oxide Synthase; D052248:Nitric Oxide Synthase Type I; D052250:Nitric Oxide Synthase Type III; D001920:Bradykinin