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-γ (IFN-γ). Pt-G and G enhanced in a concentration and time-dependent manner NO production by IFN-γ-stimulated RAW 264.7 cells. The increase of iNOS protein expression was correlated with NF-κB/DNA binding activity and occurred at transcriptional level. Pyrrolidine dithiocarbamate and N-α-para-tosyl-L-lysine chloromethyl ketone, two known inhibitors of NF-κB activation, decreased significantly NO production and iNOS protein expression as well as NF-κB/DNA binding activity. Our results show that the effect of Pt-G and G on enhancement of iNOS protein expression in IFN-γ-treated RAW 264.7 cells is mainly mediated through NF-κB and suggest that blockage of NF-κB activation reduces enhancing effect of gluten on NO production in inflamed mucosa of coeliac patients.
Similar content being viewed by others
References
Amore A, Emancipator SN, Roccatello D, Gianoglio B, Peruzzi L, Porcellini MG, Piccoli G, Coppo R (1994) Functional consequences of the binding of gliadin to cultured rat mesangial cells: bridging immunoglobulin A to cells and modulation of eicosanoid synthesis and altered cytokine production. Am J Kid Dis 23:290–301
Auricchio S, De Ritis G, De Vincenzi M, Magazzù G, Maiuri L, Mancini E (1990) Mannan and oligomers of N-acetylglucosamine protect intestinal mucosa of celiac patients with active disease from in vitro toxicity of gliadins peptides. Gastroenterology 99:973–978
Beckett CG, Dell'Olio D, Shidrawi RG, Rosen-Bronson S, Ciclitira PJ (1998) The detection and localization of inducible nitric oxide synthase production in the small intestine of patients with coeliac disease. Eur J Gastroenterol Hepat 11:529–535
Beckett CG, Dell'Olio D, Shidrawi RG, Rosen-Bronson S, Ciclitira PJ (1999) Gluten-induced nitric oxide and pro-inflammatory cytokine release by cultured coeliac small intestinal biopsies. Eur J Gastroenterol Hepatol 11:529–535
Chu SC, Marks-Konczalik J, Wu HP, Banks TC, Moss J (1998) Analysis of the cytokine-stimulated human inducible nitric oxide synthase (iNOS) gene: characterization of differences between human and mouse iNOS promoters. Biochem Biophys Res Commun 248:871–878
D'Acquisto F, de Cristofaro F, Maiuri MC, Tajana G, Carnuccio R (2001) Protective role of nuclear factor kappaB against nitric oxide-induced apoptosis in J774 macrophages. Cell Death Diff 8:144–151
Damjanov I (1987) Lectin cytochemistry and histochemistry. Lab Invest 57:5–20
De Ritis G, Occorsio P, Auricchio S, Gramenzi F, Morisi G, Silano V (1979) Toxicity of wheat flour proteins and protein-derived peptides for in vitro developing intestine from rat fetus. Pediatr Res 13:1255–1261
Drew PD, Franzoso G, Becker KG, Bours V, Carlson LM, Siebenlist U, Ozato K (1995) NFκB and interferon regulatory factor 1 physically interact and synergistically induce major histocompatibility class I gene expression. J Interferon Cytokine Res 15:1037–1045
Eberhardt W, Kunz D, Hummel R, Pfeilschifter J (1996) Molecular cloning of the rat inducible nitric oxide synthase gene promoter. Biochem Biophys Res Commun 223:752–756
Epinat J-C, Gilmore TD (1999) Diverse agent act at multiple levels to inhibit the Rel/NF-κB signal transduction pathway. Oncogene 18:6896–6909
Farré Castany MA, Kocna P, Tlaskalová-Hogenová H (1995) Binding of gliadin to lymphoblastoid, myeloid and epithelial cell lines. Folia Microbiol 40:431–435
Flohè L, Brigelius-Flohe R, Saliou C, Traber MG, Packer L (1997) Redox regulation of NF-κB activation. Free Radic Biol Med 22:1115–1126
Gao J, Morrison DC, Parmely TJ, Russell SW, Murphy WJ (1997) An interferon-gamma-activated site (GAS) is necessary for full expression of the mouse iNOS gene in response to interferon-gamma and lipopolysaccharide. J Biol Chem 272:1226–1230
Holmegren Peterson K, Fälth-Magnusson K, Magnusson K-E, Stenhammar L, Sundqvist T (1998) Children with celiac disease express inducible nitric oxide synthase in the small intestine during gluten challenge. Scand J Gastroenterol 33:939–943
Kamijo R, Harada H, Matsuyama T, Bosland M, Gerecitano J, Shapiro D, Le J, Koh SI, Kimura T, Green SJ (1994) Requirement for transcription factor IRF-1 in NO synthase induction in macrophages. Science 263:1612–1615
Kim H, Lee HS, Chang KT, Ko TH, Baek KJ, Kwon NS (1995) Chloromethyl ketones block induction of nitric oxide synthase in murine macrophages by preventing activation of nuclear factor-κB. J Immunol 154:4741–4748
Kim YM, Lee BS, Yi KY, Paik SG (1997) Upstream NF-kappaB site is required for the maximal expression of mouse inducible nitric oxide synthase gene in interferon-gamma plus lipopolysaccharide-induced RAW 264.7 macrophages. Biochem Biophys Res Commun 236:655–660
Kolberg J, Sollid LM (1985) Lectin activity of gluten identified as wheat germ agglutinin. Biochem Biophys Res Comm 130:867–872
Kontakou M, Przemioslo RT, Sturgess RP, Limb AG, Ciclitira PJ (1995) Expression of tumor necrosis factor-alfa, interleukin-6, and interleukin-2 mRNA in the jejunum of patients with coeliac disease. Scand J Gastroenterol 30:456–463
Liu RH, Hotchkiss JH (1995) Potential genotoxicity of chronically elevated nitric oxide: a review. Mutat Res 339:73–89
Maiuri L, Troncone R, Mayer M, Coletta S, Picarelli A, De Vincenzi M, Pavone V, Auricchio S (1996) In vitro activities of A-gliadin related synthetic peptides: damaging affect on the atrophic coeliac mucosa and activation of mucosal immune response in the treated coeliac mucosa. Scand J Gastroenterol 31:247–253
Marsh MN (1992) Gluten, major histocompatibility complex, and the small intestine. Gastroenterology 102:330–354
Martin E, Nathan C, Xie QW (1994) Role of interferon regulatory factor 1 in induction of nitric oxide synthase. J Exp Med 180:977–984
Marzio R, Jirillo E, Ransijn A, Mauel J, Corradin SB (1997) Expression and function of the early activation antigen CD 69 in murine macrophages. J Leukoc Biol 62:349–355
Nilsen EM, Lundin KEA, Krajci P, Scott H, Sollid LM, Brandtzaeg P (1995) Gluten specific, HLA-DQ restricted T cells from celiac mucosa produce cytokines with Th1 or Th0 profile dominated by interferon-gamma. Gut 37:766–776
Ohmori Y, Schreiber RD, Hamilton TA (1997) Synergy between interferon-γ and tumor necrosis factor-α in transcriptional activation is mediated by cooperation between signal transducer and activated of transcription 1 and nuclear factor κB. J Biol Chem 272:14899–14907
Pender SLF, Lionetti P, Murch SH, Wathan N, MacDonald TT (1996) Proteolytic degradation of intestinal mucosa extracellular matrix after lamina propria T cell activation. Gut 39:284–290
Pittschieler K, Ladinser B, Petell JK (1994) Reactivity of gliadin and lectins with celiac intestinal mucosa. Pediatr Res 36:635–641
Przemioslo R, Kontakou M, Nobili V, Ciclitira PJ (1994) Detection of interferon-gamma mRNA in the mucosa of patients with coeliac disease by in situ hybridization. Gut 35:1398–1404
Rivabene R, Mancini E, De Vincenzi M (1999) In vitro cytotoxic effect of wheat gliadin-derived peptides on the Caco-2 intestinal cell line in associated with intracellular oxidative imbalance: implications for coeliac disease. Biochem Biophys Acta 1453:152–160
Roccatello D, Amprimo MC, Coppo R, Cavalli G, Quattrocchio G, Gianoglio B, Ferrero A, di Mauro C, Sena LM, Piccoli G (1990) Influence of gluten-derived fractions on chemiluminescence production by human neutrophils. J Biolumin Chemilumin 5:161–164
Shan L, Molberg Ø, Parrot I, Hausch F, Ferda F, Gray GM, Sollid LM, Khosla C (2002) Structural basis for gluten intolerance in celiac sprue. Science 297:2275–2279
Sherman MP, Aeberhard EE, Wong VZ, Griscavage JM, Ignarro LJ (1993) Pyrrolidine dithiocarbamate inhibits induction of nitric oxide synthase activity in rat alveolar macrophages. Biochem Biophys Res Commun 191:1301–1308
Sollid LM (2000) Molecular basis of celiac disease. Ann Rev Immunol 18:53–81
Teng X, Zhang H, Snead C, Catravas J (2002) Molecular mechanisms of iNOS induction by IL-1β and IFN-γ in rat aortic smooth muscle cells. Am J Physiol Cell Physiol 282:C144–C152
Ter Steege J, Buurman W, Arends JW, Forget P (1997) Presence of inducible nitric oxide synthase, nitrotyrosine, CD68, and CD14 in the small intestine in celiac disease. Lab Invest 77:29–36
Tuckovà L, Flegelová Z, Tlaskalová-Hogenová H, Zìdek Z (2000) Activation of macrophages by food antigens: enhancing effect of gluten on nitric oxide and cytokine production. J Leukoc Biol 67:312–318
Tuckovà L, Novotna J, Novak P, Flegelová Z, Kveton T, Jelinkova L, Zìdek Z, Man P, Tlaskalová-Hogenová H (2002) Activation of macrophages by gliadin fragments: isolation and characterization of active peptide. J Leukoc Biol 71:625–631
Van Straaten EA, Koster-Kamphuis L, Bovee-Oudenhoven IM, van der Meer R, Forget P-P (1999) Increased urinary nitric oxide oxidation products in children with active coeliac disease. Acta Paediatr 88:528–531
Weisz A, Oguchi S, Cicatiello L, Esumi H (1994) Dual mechanism for the control of inducible-type NO synthase gene expression in macrophages during activation by interferon-γ and bacterial lipopolysaccharide. J Biol Chem 269:8324–8333
Weisz A, Cicatiello L, Esumi H (1996) Regulation of the mouse inducible-type nitric oxide synthase gene promoter by interferon-γ, bacterial lipopolysaccharide and NG-monomethyl-L-arginine. Biochem J 316:209–215
Xie QW, Whisnant R, Nathan C (1993) Promoter of mouse gene encoding calcium-independent nitric oxide synthase confers inducibility by interferon-γ and bacterial lipopolysaccharide. J Exp Med 177:1779–1784
Acknowledgements
This research was supported by a grant from the Italian government (PRIN 2002). We thank Prof. Hiroyasu Esumi (National Cancer Center Research Institute East, Chiba, Japan) and Prof. Alessandro Weisz (Istituto di Patologia generale e Oncologia, Seconda Università di Napoli, Naples, Italy) for their generous gift of plasmid used in this paper.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Maiuri, M.C., De Stefano, D., Mele, G. et al. Gliadin increases iNOS gene expression in interferon-γ-stimulated RAW 264.7 cells through a mechanism involving NF-κB. Naunyn-Schmiedeberg's Arch Pharmacol 368, 63–71 (2003). https://doi.org/10.1007/s00210-003-0771-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00210-003-0771-y