Interleukin-1-induced nuclear factor κB activation is inhibited by overexpression of phospholipid hydroperoxide glutathione peroxidase in a human endothelial cell line

Abstract

Oxygen radicals are commonly accepted mediators in the tumour necrosis factor-mediated nuclear factor κB (NFκB) signalling cascade, but evidence for their role during interleukin-1 (IL-1) signalling is lacking. To test the involvement of hydroperoxides we investigated whether IL-1-induced NFκB activation could be influenced by glutathione peroxidases (GPx). These enzymes remove hydroperoxides with various specificities for the hydroperoxide substrate. By overexpressing phospholipid hydroperoxide glutathione peroxidase (PHGPx), which characteristically reacts with lipophilic hydroperoxides, the roles of H2O2 and lipidhydroperoxides were assessed. A human umbilical endothelial cell line, ECV 304, was stably transfected with the genes for both PHGPx and selenophosphate synthetase (selD), which provides selenophosphate for selenoprotein biosynthesis. When grown in selenium-deficient culture medium, the double-transfected clone (ECVPHGPx+SelD+) expressed 5-fold higher (P < 0.005) PHGPx activity (measured by phosphatidylcholine hydroperoxide removal) than controls. The rate of H2O2 removal was also significantly (P < 0.01) higher in this clone. When grown with high levels of extracellular selenium (up to 100 nM selenite), PHGPx activity and H2O2 removal were enhanced substantially in control cells and transfected cells. Under these conditions, PHGPx activity was 1.7-fold (P < 0.005) higher in ECVPHGPx+SelD+, but H2O2 removal was the same as in controls. IL-1-induced NFκB activation was inhibited by selenium supplementation in control cells. In ECVPHGPx+SelD+ under conditions of selenium restriction, IL-1 induced NFκB activation only to a similar extent as under conditions of selenium supplementation in controls, and activation was abolished with 50 nM sodium selenite. These results show that overexpressed PHGPx is sufficient to inhibit NFκB activation, and suggests that NFκB activation by IL-1 is mediated by a preferential substrate of PHGPx, such as a fatty acid hydroperoxide, rather than by H2O2, the preferred substrate of the more abundant cytosolic GPx.

Self-regulatory role of 4-hydroxynonenal in signaling for stress-induced programmed cell death

“…The enzymes such as glutathione S-transferases, aldehyde dehydrogenases, and aldose reductases and transporters such as RalBP1 and MRP1 that catalyze the efflux of its metabolites regulate its intracellular concentrations [2][3][4][5][6][7][8][9][10][11][12][13][14][15]. GSTs catalyze the conjugation of HNE to glutathione (GSH) which is the major pathway for disposition of HNE.…”

“…These studies have opened a new area in the field of ROS-induced signaling focusing on the regulatory roles of the enzymes involved in the formation and metabolism of HNE. Recent studies in this area have shown that enzymes such as glutathione S-transferases (GSTs), aldehyde dehydrogenases, aldose reductase, glutathione peroxidase, and RalBP1 (Ral-binding protein 1) that are among the major determinants of intracellular levels of HNE can modulate stress-induced signaling for programmed cell death [2][3][4][5][6][7][8][9][10]. Some of these studies [11][12][13][14] seem to have major clinical implications particularly in regard to cancer chemotherapy [11,12], inhibition of tumor growth [13], and sepsis [14] as discussed briefly later in this review.…”

Inhibition of NF-κB DNA binding and nitric oxide induction in human T cells and lung adenocarcinoma cells by selenite treatment

“…Reduced cysteine residue(s) of NF-B appear to be required for its actual binding to the promotor region of a target gene (11). Dependence of the early activation steps of this cascade on reactive oxygen species is indicated by the inhibitory effects of added N-acetylcysteine and metal chelators (9,10) and increased intracellular levels of phospholipid hydroperoxide glutathione peroxidase (12).…”

Distinct Promoters Determine Alternative Transcription of gpx-4 into Phospholipid-Hydroperoxide Glutathione Peroxidase Variants

“…It was therefore considered to synergistically act with vitamin E in protecting membranes from lipid peroxidation (9). More recently, however, PHGPx has been implicated in regulating the synthesis of lipid mediators by silencing 5-and 15-lipoxygenase (10 -12), in suppressing hydroperoxideinduced apoptosis (13), in dampening cytokine-induced gene activation (14), in peroxynitrite scavenging (15), and in spermatogenesis (16).…”