Regulation of selenoprotein GPx4 expression and activity in human endothelial cells by fatty acids, cytokines and antioxidants


      Phospholipid hydroperoxide glutathione peroxidase (GPx4) is the only antioxidant enzyme known to directly reduce phospholipid hydroperoxides within membranes and lipoproteins, acting in conjunction with α-tocopherol to inhibit lipid peroxidation. Peroxidation of lipids has been implicated in a number of pathophysiological processes, including inflammation and atherogenesis. We investigated the relative positive and negative effects of specific polyunsaturated fatty acids (PUFAs) and inflammatory cytokines on the activity and gene expression of the selenium-dependant redox enzyme GPx4. In human umbilical vein endothelial cells (HUVEC), GPx4 mRNA levels and activity were increased optimally by 114 nM selenium (as sodium selenite). Docosahexaenoic acid (DHA) and conjugated linoleic acid (CLA) further increased mRNA levels whereas arachidonic acid (ARA) had no effect; enzyme activity was decreased by DHA, was unaffected by CLA or was increased by ARA. GPx4 protein levels increased with selenium, ARA and DHA addition but not with CLA. Interleukin-1β (IL-1β) increased GPx4 mRNA, protein and activity whereas TNFα at 1 ng/ml increased activity while at 3 ng/ml it reduced activity and mRNA. Conversely, α-tocopherol reduced mRNA levels without affecting activity. These results indicate that lipids, cytokines and antioxidants modulate GPx4 in a complex manner that in the presence of adequate selenium, may favour protection against potentially proatherogenic processes.


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        • Arai M.
        • Imai H.
        • Koumura T.
        • Yoshida M.
        • Emoto K.
        • Umeda M.
        • et al.
        Mitochondrial phospholipid hydroperoxide glutathione peroxidase plays a major role in preventing oxidative injury to cells.
        J. Biol. Chem. 1999; 274: 4924-4933
        • Thomas J.P.
        • Geiger P.G.
        • Girotti A.W.
        Lethal damage to endothelial cells by oxidized low density lipoprotein: role of selenoperoxidases in cytoprotection against lipid hydroperoxide- and iron-mediated reactions.
        J. Lipid Res. 1993; 34: 479-490
        • Lindau-Shepard B.
        • Shaffer J.B.
        • Del Vecchio P.J.
        Overexpression of manganous superoxide dismutase (MnSOD) in pulmonary endothelial cells confers resistance to hyperoxia.
        J. Cell. Physiol. 1994; 161: 237-242
        • Lu D.
        • Maulik N.
        • Moraru I.I.
        • Kreutzer D.L.
        • Das D.K.
        Molecular adaptation of vascular endothelial cells to oxidative stress.
        Am. J. Physiol. 1993; 264: C715-722
        • Cross C.E.
        • Halliwell B.
        • Borish E.T.
        • Pryor W.A.
        • Ames B.N.
        • Saul R.L.
        • et al.
        Oxygen radicals and human disease.
        Ann. Intern. Med. 1987; 107: 526-545
        • Aviram M.
        Review of human studies on oxidative damage and antioxidant protection related to cardiovascular diseases.
        Free Radic. Res. 2000; 33: S85-97
        • Brigelius-Flohe R.
        • Friedrichs B.
        • Maurer S.
        • Streicher R.
        Determinants of GPx4 expression in a cultured endothelial cell line.
        Biomed. Environ. Sci. 1997; 10: 163-176
        • Dolecek T.A.
        Epidemiological evidence of relationships between dietary polyunsaturated fatty acids and mortality in the multiple risk factor intervention trial.
        Proc. Soc. Exp. Biol. Med. 1992; 200: 177-182
        • Kromhout D.
        • Bosschieter E.B.
        • de Lezenne Coulander C.N.
        The inverse relation between fish consumption and 20-year mortality from coronary heart disease.
        Eng. J. Med. 1985; 312: 1205-1209
        • Blonk M.C.
        • Bilo H.J.
        • Nauta J.J.
        • Popp-Snijders C.
        • Mulder C.
        • Donker A.J.
        Dose-response effects of fish-oil supplementation in healthy volunteers.
        Am. J. Clin. Nutr. 1990; 52: 120-127
        • Kinsella J.E.
        • Lokesh B.
        • Stone R.A.
        Dietary n-3 polyunsaturated fatty acids and amelioration of cardiovascular disease: possible mechanisms.
        Am. J. Clin. Nutr. 1990; 52: 1-28
        • Crosby A.J.
        • Wahle K.W.
        • Duthie G.G.
        Modulation of glutathione peroxidase activity in human vascular endothelial cells by fatty acids and the cytokine interleukin-1 beta.
        Biochim. Biophys. Acta. 1996; 1303: 187-192
        • Wahle K.W.
        • Rotondo D.
        Fatty acids and endothelial cell function: regulation of adhesion molecule and redox enzyme expression.
        Curr. Opin. Clin. Nutr. Metab. Care. 1999; 2: 109-115
        • Collie-Duguid E.S.
        • Wahle K.W.
        Inhibitory effect of fish oil N-3 polyunsaturated fatty acids on the expression of endothelial cell adhesion molecules.
        Biochem. Biophys. Res. Commun. 1996; 220: 969-974
        • Weber C.
        • Erl W.
        • Pietsch A.
        • Danesch U.
        • Weber P.C.
        Docosahexaenoic acid selectively attenuates induction of vascular cell adhesion molecule-1 and subsequent monocytic cell adhesion to human endothelial cells stimulated by tumor necrosis factor-alpha.
        Arterioscler. Thromb. Vasc. Biol. 1995; 15: 622-628
        • Mayer K.
        • Merfels M.
        • Muhly-Reinholz M.
        • Gokorsch S.
        • Rosseau S.
        • Lohmeyer T.
        • et al.
        Omega-3 fatty acids suppress monocyte adhesion to human endothelial cells: role of endothelial PAF generation.
        Am. J. Physiol. Heart. Circ. Physiol. 2002; 283: H811-818
        • Maiorino M.
        • Thomas J.P.
        • Girotti A.W.
        • Ursini F.
        Reactivity of phospholipid hydroperoxide glutathione peroxidase with membrane and lipoprotein lipid hydroperoxides.
        Free Radic. Res. Commun. 1991; 12–13: 131-135
        • Funk C.D.
        Prostaglandins and leukotrienes: advances in eicosanoid biology.
        Science. 2001; 294: 1871-1875
      1. Flohe L, Wingender E, Brigelius-Flohe R. The regulation of glutathione peroxidases. In: Forman H, Cadenas E, editors. Oxidative stress and signal transduction. New York: Chapman & Hall; 1997. p. 415–40.

      2. Sambrook J, Fritsch EF, Maniatis T, editors. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press; 1989.

        • Weitzel F.
        • Ursini F.
        • Wendel A.
        Phospholipid hydroperoxide glutathione peroxidase in various mouse organs during selenium deficiency and repletion.
        Biochim. Biophys. Acta. 1990; 1036: 88-94
        • Olsen O.E.
        • Palmer I.S.
        • Carey H.H.
        Modification of the official fluorometric method for selenium assay in plants.
        J. Assoc. Off. Anal. Chem. 1975; 58: 117-121
        • De Leenheer A.P.
        • De Bevere V.O.
        • Cruyl A.A.
        • Claeys A.E.
        Determination of serum alpha-tocopherol (vitamin E) by high-performance liquid chromatography.
        Clin. Chem. 1978; 24: 585-590
        • Leist M.
        • Raab B.
        • Maurer S.
        • Rosick U.
        • Brigelius-Flohe R.
        Conventional cell culture media do not adequately supply cells with antioxidants and thus facilitate peroxide-induced genotoxicity.
        Free Radic. Biol. Med. 1996; 21: 297-306
        • Brown K.M.
        • Pickard K.
        • Nicol F.
        • Beckett G.J.
        • Duthie G.G.
        • Arthur J.R.
        Effects of organic and inorganic selenium supplementation on selenoenzyme activity in blood lymphocytes, granulocytes, platelets and erythrocytes.
        Clin. Sci. (Lond.). 2000; 98: 593-599
        • Bermano G.
        • Arthur J.R.
        • Hesketh J.E.
        Selective control of cytosolic glutathione peroxidase and phospholipid hydroperoxide glutathione peroxidase mRNA stability by selenium supply.
        FEBS Lett. 1996; 387: 157-160
        • Wingler K.
        • Bocher M.
        • Flohe L.
        • Kollmus H.
        • Brigelius-Flohe R.
        mRNA stability and selenocysteine insertion sequence efficiency rank gastrointestinal glutathione peroxidase high in the hierarchy of selenoproteins.
        Eur J Biochem. 1999; 259: 149-157
        • Hara S.
        • Shoji Y.
        • Sakurai A.
        • Yuasa K.
        • Himeno S.
        • Imura N.
        Effects of selenium deficiency on expression of selenoproteins in bovine arterial endothelial cells.
        Biol. Pharm. Bull. 2001; 24: 754-759
        • Bermano G.
        • Nicol F.
        • Dyer J.A.
        • Sunde R.A.
        • Beckett G.J.
        • Arthur J.R.
        • et al.
        Tissue-specific regulation of selenoenzyme gene expression during selenium deficiency in rats.
        Biochem. J. 1995; 311: 425-430
        • Roveri A.
        • Maiorino M.
        • Nisii C.
        • Ursini F.
        Purification and characterization of phospholipid hydroperoxide glutathione peroxidase from rat testis mitochondrial membranes.
        Biochim Biophys Acta. 1994; 1208: 211-221
        • Lee K.N.
        • Kritchevsky D.
        • Pariza M.W.
        Conjugated linoleic acid and atherosclerosis in rabbits.
        Atherosclerosis. 1994; 108: 19-25
        • Dong Z.M.
        • Chapman S.M.
        • Brown A.A.
        • Frenette P.S.
        • Hynes R.O.
        • Wagner D.D.
        The combined role of P- and E-selectins in atherosclerosis.
        J. Clin. Invest. 1998; 102: 145-152
        • Collins R.G.
        • Velji R.
        • Guevara N.V.
        • Hicks M.J.
        • Chan L.
        • Beaudet A.L.J.
        P-Selectin or intercellular adhesion molecule (ICAM)-1 deficiency substantially protects against atherosclerosis in apolipoprotein E-deficient mice.
        Exp. Med. 2000; 191: 189-194
        • Huang H.S.
        • Chang W.C.
        • Chen C.J.
        Involvement of reactive oxygen species in arsenite-induced downregulation of phospholipid hydroperoxide glutathione peroxidase in human epidermoid carcinoma A431 cells.
        Free Radic. Biol. Med. 2002; 33: 864-873
        • Marui N.
        • Offermann M.K.
        • Swerlick R.
        • Kunsch C.
        • Rosen C.A.
        • Ahmad M.
        • et al.
        Vascular cell adhesion molecule-1 (VCAM-1) gene transcription and expression are regulated through an antioxidant-sensitive mechanism in human vascular endothelial cells.
        J. Clin. Invest. 1993; 92: 1866-1874
        • Brigelius-Flohe R.
        • Friedrichs B.
        • Maurer S.
        • Schultz M.
        • Streicher R.
        Interleukin-1-induced nuclear factor kappa B activation is inhibited by overexpression of phospholipid hydroperoxide glutathione peroxidase in a human endothelial cell line.
        Biochem. J. 1997; 328: 199-200