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Gliadin increases iNOS gene expression in interferon-γ-stimulated RAW 264.7 cells through a mechanism involving NF-κB

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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.

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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.

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Authors and Affiliations

  1. Dipartimento di Farmacologia Sperimentale, Facoltà di Scienze Biotecnologiche, Università degli Studi di Napoli "Federico II", Via D. Montesano 49, 80131, Naples, Italy

    Maria Chiara Maiuri, Daniela De Stefano, Guido Mele & Rosa Carnuccio

  2. Dipartimento di Biochimica e Biotecnologie Mediche, Facoltà di Scienze Biotecnologiche, Università degli Studi di Napoli "Federico II", Via S. Pansini 5, 80131, Naples, Italy

    Barbara Iovine & Maria Assunta Bevilacqua

  3. Dipartimento di Pediatria, Facoltà di Medicina e Chirurgia, Università degli Studi di Napoli "Federico II", Via S. Pansini 5, 80131, Naples, Italy

    Luigi Greco & Salvatore Auricchio

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  1. Maria Chiara Maiuri
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Correspondence to Rosa Carnuccio.

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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

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  • DOI: https://doi.org/10.1007/s00210-003-0771-y

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