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Docosahexaenoic acid enhances the antioxidant response of human fibroblasts by upregulating γ-glutamyl-cysteinyl ligase and glutathione reductase

Published online by Cambridge University Press:  08 March 2007

Khelifa Arab ,
Adrien Rossary ,
Françoise Flourié ,
Yves Tourneur  and
Jean-Paul Steghens
Khelifa Arab
Affiliation:
UF 21455 Oxidative Stress and Vitamins,Biochemistry Federation, E. Herriot Hospital, Lyon, France EA 3090, Claude Bernard University Lyon 1, France
Adrien Rossary
Affiliation:
UF 21455 Oxidative Stress and Vitamins,Biochemistry Federation, E. Herriot Hospital, Lyon, France EA 3090, Claude Bernard University Lyon 1, France
Françoise Flourié
Affiliation:
UF 21455 Oxidative Stress and Vitamins,Biochemistry Federation, E. Herriot Hospital, Lyon, France
Yves Tourneur
Affiliation:
Quantimetry Center, Pharmacy Faculty, Claude Bernard University Lyon 1, France
Jean-Paul Steghens*
Affiliation:
UF 21455 Oxidative Stress and Vitamins,Biochemistry Federation, E. Herriot Hospital, Lyon, France EA 3090, Claude Bernard University Lyon 1, France
*
*Corresponding author: Dr Jean-Paul Steghens, fax +33 4 72 11 06 75, email jean-paul.steghens@chu-lyon.fr
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Abstract

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The chemopreventive effects of dietary n-3 PUFA in various pathologies has so far remained controversial, and we were interested in studying their potential influence on cell redox status. DHA (22:6n-3), a typical highly unsaturated n-3 PUFA, was used at 30µmol/l in a model of human fibroblast cell culture. A dose–response effect, roughly linear, was checked for DHA between 0 and 60µmol/l, and was accompanied by a large increase in intracellular GSH content. A time course study of this effect shows that, after a short fall, as soon as 4h after the beginning of the experiment, the large increase in the GSH content was associated with elevated catalytic activities of γ-glutamyl-cysteinyl ligase, glutathione reductase and glutathione S-transferase. This coordinated response is characteristic of an antioxidant response and was confirmed by the induction of expression of mRNA for γ-glutamyl-cysteinyl ligase, glutathione reductase and haem-oxygenase. This large increase in the GSH content contributes to decreasing the reactive oxygen species level, as assessed by the decreased accumulation of dichlorofluorescein inside cells. To our knowledge, this is the first report on a specific and potent effect of DHA for decreasing the oxidative stress of human fibroblasts.

Keywords

Docosahexaenoic acidChemopreventionThiol-redox statusFibroblastsGlutathioneOxidative stress
Type
Research Article
Information
British Journal of Nutrition , Volume 95 , Issue 1 , January 2006 , pp. 18 - 26
Copyright
Copyright © The Nutrition Society 2006

References

Akihisa, T, Tokuda, H, Ogata, M, Ukiya, M, Iizuka, M, Suzuki, T, Metori, K, Shimizu, N & Nishino, HCancer chemopreventive effects of polyunsaturated fatty acids. Cancer Lett (2004) 205, 9–13.CrossRefGoogle ScholarPubMed
Alam, J & Cook, JLTranscriptional regulation of the heme oxygenase-1 gene via the stress response element pathway. Curr Pharm Des (2003) 9 2499–2511.CrossRefGoogle ScholarPubMed
Anderson, MEGlutathione and glutathione delivery compounds. Avd Pharmacol (1997) 38, 65–78.Google ScholarPubMed
Arab, K & Steghens, JPPlasma lipid hydroperoxides measurement by an automated xylenol orange method. Anal Biochem (2004) 325, 158–163.CrossRefGoogle ScholarPubMed
Barker, JE, Heales, SJR, Cassidy, A, Bolanos, JP, Land, JM & Clark, JBDepletion of brain glutathione results in a decrease of glutathione reductase activity; an enzyme susceptible to oxidative damage. Brain Res (1996) 716, 118–122.CrossRefGoogle Scholar
Brown, ER & Subbaiah, PVDifferential effects of eicosapentanoic acid and docosahexaenoic acid on human skin fibroblasts. Lipids (1994) 29, 825–829.CrossRefGoogle ScholarPubMed
Carter, WO, Narayanan, PK & Robinson, JPIntracellular hydrogen peroxide and superoxide anion detection in endothelial cells. J Leukoc Bio (1994) 55 253–258.CrossRefGoogle ScholarPubMed
Chik, K, Flourie, F, Arab, K & Steghens, JPA simple and fast kinetic LC/MS measurement of γ glutamyl-cysteinyl ligase activity. J Chromatogr B, (2005) May 16. Available online. DOI:10.101/.jchromb.2004.04.040.CrossRefGoogle Scholar
Davidson, LA, Nguyen, DV, Hokanson, RM et al. Chemopreventive n–3 polyunsaturated fatty acids reprogram genetic signatures during colon cancer initiation and progression in the rat. Cancer Res (2004) 64, 6797–6804.CrossRefGoogle ScholarPubMed
Dickinson, DA, Iles, N, Watanabe, KE, Iwamoto, T, Zhang, H, Krzywanski, DM & Forman, HJ4–Hydroxynonenal induces glutamate cysteine ligase through JNK in HBE1 cells. Free Radic Biol Med (2002) 33, 974–987.CrossRefGoogle ScholarPubMed
Germain, E, Chajes, V, Cognault, S, Lhuillery, C & Bougnoux, PEnhancement of doxorubicin cytotoxicity by polyunsaturated fatty acids in the human breast tumor cell line MDA-MB-231:relationship to lipid peroxidation. Int J Cancer (2002) 75, 578–583.3.0.CO;2-5>CrossRefGoogle Scholar
Habig, WH, Pabst, MJ & Jakoby, WBGlutathione S–transferases. The first enzymatic step in mercapturic acid formation. J Biol Chem (1974) 29, 249–7139.Google Scholar
Kim, H, Akbar, M, Lau, A & Esdall, LInhibition of neuronal apoptosis by docosahexaenoic acid (22:6n–3). J Biol Chem (2000) 275, 35215–35223.CrossRefGoogle ScholarPubMed
Komatsu, W, Ishihara, K, Murata, M, Saito, H & Shinohara, KDocosahexaenoic acid suppresses nitric oxide production and inducible nitric oxide synthase expression in interferon–gamma plus lipopolysaccharide–stimulated murine macrophages by inhibiting the oxidative stress. Free Radic Biol Med (2003) 34, 1006–1016.CrossRefGoogle ScholarPubMed
Kris-Etherton, PM, Harris, WS & Appel, LJFish consumption, fish oil, omega-3 fatty acids, and cardiovascular disease. Circulation (2002) 106, 2747–2757.CrossRefGoogle ScholarPubMed
Larsson, SC,Kumlin, M, Ingelman—Sundberg, M & Wolk, ADietary long-chain n-3 fatty acids for the prevention of cancer: a review of potential mechanisms Am J Clin Nutr (2004) 79,935–945.CrossRefGoogle Scholar
Lee, JM, Calkins, MJ,Chan, K,Kan, YW & Johnson, JAIdentification of the NF-E2-related factor-2-dependent genes conferring protection against oxidative stress in primary cortical astrocytes using oligonucleotide microarray analysis J Biol Chem (2003a) 278 12029–12038.CrossRefGoogle ScholarPubMed
Lee, HT,Xu, H, Ota-Setlik, A & Emala, CWOxidant preconditioning protects human proximal tubular cells against lethal oxidant injury via p38 MAPK and heme oxygenase-1. Am J Nephrol (2003b) 23, 324–333.CrossRefGoogle ScholarPubMed
Levonen, AL, Landar, A, Ramachandran, A, Ceaser, EK,Dickinson, DA,Zanoni, G,Morrow, JD & Darley-Usmar, VMCellular mechanisms of redox cell signalling: role of cysteine modification in controlling antioxidant defences in response to electrophilic lipid oxidation products. Biochem J (2004) 378, 373–382.CrossRefGoogle ScholarPubMed
Liu, W,Kato, M, Akhand, AA,Hayakawa, A,Suzuki, H,Miyata, T,Kurokawa, K,Hotta, Y,Ishikawa, N & Nakashima, I4-hydroxynonenal induces a cellular redox status-related activation of the caspase cascade for apoptotic cell death J Cell Sci (2000) 113, 635–641.CrossRefGoogle ScholarPubMed
Locus Link http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?CMD=search&DB=gene (2005)Google Scholar
Lopez-Garcia, E, Schulze, MZ,Manson, JE,Meigs, JB,Albert, CM,Rifai, N,Willet, WC & Hu, FBConsumption of (n-3) fatty acids is related to plasma biomarkers of inflammation and endothelial activation in women. J Nutr (2004) 134, 1806–1811.CrossRefGoogle Scholar
Marcheselli, LV, Hong, S, Lukiw, JW.et al Novel docosanoids inhibit brain ischemia-reperfusion mediated leukocyte infiltration and pro-inflammatory gene expression J Biol Chem (2003) 278 43807–43817.CrossRefGoogle ScholarPubMed
Martinez, M, Vazquez, E, Garcia-Silva, MT,Manzanares, J,Bertran, JM,Castello, F & Mougan, ITherapeutic effects of docosahexaenoic acid ethyl ester in patients with generalized peroxisomal disorders. J Clin Nutr, (2000) 71 Suppl., 376S–385S.CrossRefGoogle ScholarPubMed
Merendino, N, Molinari, R,Loppi, B,Pessina, G,D'Aquino, M,Tomassi, G & Velloti, FInduction of apoptosis in human pancreatic cancer cell by docosahexaenoic acid. Ann N Y Acad Sci (2003) 1010 361–364.CrossRefGoogle ScholarPubMed
Primer3 software http://cbr-rbc.nrc-cnrc.gc.ca/cgi-bin/primer3_www.cgi (2005)Google Scholar
Rhodes, LE, Shanbakhti, H, Azurdia, RM,et al Effect of eicosapentanoic acid, an omega-3 polyunsaturated fatty acid, on UVR-related cancer risk in humans. An assessment of early genotoxic markers. Carcinogenesis (2003) 24, 919–925.CrossRefGoogle ScholarPubMed
Romero-Ramos, M, Venero, JL, Garcia-Rodriguez, S,Ayala, A,Machado, A & Cano, JSemichronic inhibition of glutathione reductase promotes oxidative damage to proteins and induces both transcription and translation of tyrosine hydroxylase in the nigrostriatal system. Free Radic Res (2003) 37, 1003–1012.CrossRefGoogle ScholarPubMed
Rossi, R,Milzani, A,Dalle-Donne, I,Giustarini, D,Lusini, L,Colombo, R & Di Simplicio, PBlood glutathione disulfide: in vivo factor or in vitro artifact?" Clin Chem. (2002) 48,742– 753.CrossRefGoogle ScholarPubMed
Rushmore, TH,Morton, MR & Pickett, CBThe antioxidant responsive element. Activation by oxidative stress and identifi-cation of the DNA consensus sequence required for functional activity. J Biol Chem (1991) 266,11632–11639.CrossRefGoogle Scholar
Shi, MM, Kugelman, A, Iwalmoto, T,Tian, L & Forman, HJQuinone-induced oxidative stress elevated glutathione and induces g-glutamylcysteine synthetase activity in rat lung epithelial L2 cells. J Biol Chem (1994) 269,26512–26517.CrossRefGoogle Scholar
Steghens, JP,Flourie, F, Arab, K &Collombel, CFast liquid chromatography-mass spectrometry glutathione measurement in whole blood: micromolar GSSG is a sample preparation artifact. J Chromatogr B (2003) 25, 343–349.CrossRefGoogle Scholar
Wasserman, WW & Fahl, WEFunctional antioxidant responsive elements Proc Natl Acad Sci (1997) 94, 5361–5366.CrossRefGoogle ScholarPubMed
Wen, B, Deutsch, E,Opolon, P,Frascogna, V,Connault, E & Bourhis, n-3 polyunsaturated fatty acids decrease mucosal/epidermal reactions and enhance antitumour effect of ionising radiation with inhibition of tumour angiogenesis Br J Cancer (2003) 89, 1102–1107.CrossRefGoogle ScholarPubMed
Wild, AC, Moinova, HR & Mulcahy, TRegulation of g-glutamylcysteine synthetase subunit gene expression by the transcription factor Nrf2. J Biol Chem (1999) 274,33627–33636.CrossRefGoogle Scholar
Worthington, DJ & Rosemeyer, MAHuman glutathione reductase: purification of the crystalline from erythrocytes Eur J Biochem (1974) 48,166–177.CrossRefGoogle ScholarPubMed
Wu, G, Fang, YZ, Yang, S,Lupton, JR & Turner, NDGlutathione metabolism and its implications for health. J Nutr (2004) 134, 489–492.CrossRefGoogle ScholarPubMed
Ziboh, AV, Miller, CC & Cho, YMetabolism of polyunsaturated fatty acids by skin epidermal enzymes: generation of antiinflammatory and antiproliferative metabolites. Am J Clin Nutr (2000) 71, Suppl., 361S–366S.CrossRefGoogle ScholarPubMed