MURAL - Maynooth University Research Archive Library



    Lipid Peroxidation Is Involved in the Activation of NF-kB by Tumor Necrosis Factor but Not Interleukin-1 in the Human Endothelial Cell Line ECV304


    Bowie, Andrew G. and Moynagh, Paul N. and O'Neill, Luke A.J. (1997) Lipid Peroxidation Is Involved in the Activation of NF-kB by Tumor Necrosis Factor but Not Interleukin-1 in the Human Endothelial Cell Line ECV304. Journal of Biological Chemistry, 272 (41). pp. 25941-25950. ISSN 0021-9258

    [img]
    Preview
    Download (664kB) | Preview


    Share your research

    Twitter Facebook LinkedIn GooglePlus Email more...



    Add this article to your Mendeley library


    Abstract

    It has been proposed that reactive oxygen species, and in particular H2O2, may be involved in the activation of NF-kB by diverse stimuli in different cell types. Here we have investigated the effect of a range of putative antioxidants on NF-kB activation by interleukin-1 and tumor necrosis factor as well as the ability of H2O2 to activate NF-kB in primary human umbilical vein endothelial cells and the transformed human endothelial cell line ECV304. Activation of NF-kB and stimulation of IkBa degradation by H2O2 was only evident in the transformed cells and required much longer contact times than that observed with interleukin-1 or tumor necrosis factor. Furthermore, only H2O2 was sensitive to Nacetyl- L-cysteine, and no increase in H2O2 was detected in response to either cytokine. Pyrrolidine dithiocarbamate has been purported to be a specific antioxidant inhibitor of NF-kB that acts independently of activating agent or cell type. However, we found that tumor necrosis factor- but not interleukin-1-driven NF-kB activation and IkBa degradation were sensitive to pyrrolidine dithiocarbamate in transformed cells, while neither pathway was inhibited in primary cells. Phorbol ester-mediated activation was sensitive in both transformed and primary cells. Other antioxidants failed to inhibit either cytokine, while the iron chelators desferrioxamine and 2,2,6,6-tetramethylpiperidine-1-oxyl mimicked the pattern of inhibition seen for the dithiocarbamate. This suggested that pyrrolidine dithiocarbamate was inhibiting NF-kB activation in endothelial cells primarily through its iron-chelating properties. Tumor necrosis factor, but not interleukin-1, was found to induce lipid peroxidation in ECV304 cells. This was inhibited by pyrrolidine dithiocarbamate and desferrioxamine. t-Butyl hydroperoxide, which induces lipid peroxidation, activated NF-kB. Finally, butylated hydroxyanisole, which inhibits lipid peroxidation but has no iron-chelating properties, inhibited NF-kB activation by tumor necrosis factor but not interleukin-1. Taken together, the results argue against a role for H2O2 in NF-kB activation by cytokines in endothelial cells. Furthermore, tumor necrosis factor and interleukin- 1 activate NF-kB through different mechanisms in ECV304 cells, with the tumor necrosis factor pathway involving iron-catalyzed lipid peroxidation.

    Item Type: Article
    Additional Information: This research was originally published in the Journal of Biological Chemistry. Bowie, AG, Moynagh, P.N. and O'Neill LA. (1997) 'Lipid peroxidation is involved in the activation of NF-kappaB by tumor necrosis factor but not interleukin-1 in the human endothelial cell line ECV304. Lack of involvement of H2O2 in NF-kappaB activation by either cytokine in both primary and transformed endothelial cells'. The Journal of Biological Chemistry, 272 :25941-25950
    Keywords: Lipid Peroxidation; NF-kB; Tumor Necrosis Factor; Interleukin-1; Human Endothelial Cell Line ECV304;
    Academic Unit: Faculty of Science and Engineering > Biology
    Item ID: 7194
    Identification Number: https://doi.org/10.1074/jbc.272.41.25941
    Depositing User: Professor Paul Moynagh
    Date Deposited: 18 Jul 2016 09:10
    Journal or Publication Title: Journal of Biological Chemistry
    Publisher: American Society for Biochemistry and Molecular Biology
    Refereed: Yes
    Funders: Forbairt, Health Research Board (HRB)
    URI:

    Repository Staff Only(login required)

    View Item Item control page

    Downloads

    Downloads per month over past year