title: Gliadin increases iNOS gene expression in interferon-gamma-stimulated RAW 264.7 cells through a mechanism involving NF-kappa B.
abstract: Nitric oxide (NO) plays an important role in the pathogenesis of the histological changes seen in coeliac disease. We have investigated the effect of peptic-tryptic digest of gliadin (Pt-G) and gliadin (G) on inducible nitric oxide synthase (iNOS) protein expression in RAW 264.7 macrophages stimulated with interferon-gamma (IFN-gamma). Pt-G and G enhanced in a concentration and time-dependent manner NO production by IFN-gamma-stimulated RAW 264.7 cells. The increase of iNOS protein expression was correlated with NF-kappa B/DNA binding activity and occurred at transcriptional level. Pyrrolidine dithiocarbamate and N-alpha-para-tosyl-L-lysine chloromethyl ketone, two known inhibitors of NF-kappa B activation, decreased significantly NO production and iNOS protein expression as well as NF-kappa B/DNA binding activity. Our results show that the effect of Pt-G and G on enhancement of iNOS protein expression in IFN-gamma-treated RAW 264.7 cells is mainly mediated through NF-kappa B and suggest that blockage of NF-kappa B activation reduces enhancing effect of gluten on NO production in inflamed mucosa of coeliac patients.
authors.affiliation: Dipartimento di Farmacologia Sperimentale, Facoltà di Scienze Biotecnologiche, Università degli Studi di Napoli Federico II, Via D. Montesano 49, 80131 Naples, Italy.
authors.forename: Maria Chiara
authors.initials: MC
authors.lastname: Maiuri
authors.forename: Daniela
authors.initials: D
authors.lastname: De Stefano
authors.forename: Guido
authors.initials: G
authors.lastname: Mele
authors.forename: Barbara
authors.initials: B
authors.lastname: Iovine
authors.forename: Maria Assunta
authors.initials: MA
authors.lastname: Bevilacqua
authors.forename: Luigi
authors.initials: L
authors.lastname: Greco
authors.forename: Salvatore
authors.initials: S
authors.lastname: Auricchio
authors.forename: Rosa
authors.initials: R
authors.lastname: Carnuccio
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D004586:Electrophoresis; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D015870:Gene Expression; D005903:Gliadin / Q000494:pharmacology*; D007371:Interferon-gamma / Q000378:metabolism* / Q000494:pharmacology; D008264:Macrophages / Q000378:metabolism; D051379:Mice; D016328:NF-kappa B / Q000378:metabolism*; D009569:Nitric Oxide / Q000378:metabolism; D019001:Nitric Oxide Synthase / Q000096:biosynthesis* / Q000235:genetics; D052247:Nitric Oxide Synthase Type II; D009573:Nitrites / Q000378:metabolism; D011759:Pyrrolidines / Q000494:pharmacology; D013859:Thiocarbamates / Q000494:pharmacology; D014107:Tosyllysine Chloromethyl Ketone / Q000494:pharmacology
chemical_list: D004791:Enzyme Inhibitors; D016328:NF-kappa B; D009573:Nitrites; D011759:Pyrrolidines; D013859:Thiocarbamates; D014107:Tosyllysine Chloromethyl Ketone; C020972:pyrrolidine dithiocarbamic acid; D009569:Nitric Oxide; D007371:Interferon-gamma; D005903:Gliadin; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; C496317:Nos2 protein, mouse