title: Nrf2 deficiency causes lipid oxidation, inflammation, and matrix-protease expression in DHA-supplemented and UVA-irradiated skin fibroblasts.
abstract: Fish oil rich in docosahexaenoic acid (DHA) has beneficial effects on human health. Omega-3 polyunsaturated fatty acids are precursors of eicosanoids and docosanoids, signaling molecules that control inflammation and immunity, and their dietary uptake improves a range of disorders including cardiovascular diseases, ulcerative colitis, rheumatoid arthritis, and psoriasis. The unsaturated nature of these fatty acids, however, makes them prone to oxidation, especially when they are incorporated into (membrane) phospholipids. The skin is an organ strongly exposed to oxidative stress, mainly due to solar ultraviolet radiation. Thus, increased levels of PUFA in combination with oxidative stress could cause increased local generation of oxidized lipids, whose action spectrum reaches from signaling molecules to reactive carbonyl compounds that can crosslink biomolecules. Here, we investigated whether PUFA supplements to fibroblasts are incorporated into membrane phospholipids and whether an increase of PUFA within phospholipids affects the responses of the cells to UV exposure. The redox-sensitive transcription factor Nrf2 is the major regulator of the fibroblast stress response to ultraviolet radiation or exposure to oxidized lipids. Here we addressed how Nrf2 signaling would be affected in PUFA-supplemented human dermal fibroblasts and mouse dermal fibroblasts from Nrf2-deficient and wild type mice. We found, using HPLC-tandem MS, that DHA supplements to culture media of human and murine fibroblasts were readily incorporated into phospholipids and that subsequent irradiation of the supplemented cells with UVA resulted in an increase in 1-palmitoyl-2-(epoxyisoprostane-E2)-sn-glycero-3-phosphorylcholine and Oxo-DHA esterified to phospholipid, both of which are Nrf2 agonists. Also, induction of Nrf2 target genes was enhanced in the DHA-supplemented fibroblasts after UVA irradiation. In Nrf2-deficient murine fibroblasts, the expression of the target genes was, as expected, decreased, but surprisingly, expression of TNFα and MMP13 was strongly induced in DHA-supplemented, UVA-irradiated cells. Also, Nrf2-deficient cells had increased levels of oxidized phospholipids relative to the unoxidized precursors after UVA irradiation. Our data suggest that under ultraviolet stress a functioning Nrf2 system is required to prevent DHA-induced inflammation and matrix degradation in dermal fibroblasts.
authors.affiliation: Department of Dermatology, Medical University of Vienna, Anna Spiegel Gebäude E6 Lab5, 1090 Vienna, Austria; Christian Doppler Laboratory for Biotechnology of Skin Aging, Vienna, Austria. Electronic address: florian.gruber@meduniwien.ac.at.
authors.forename: Florian
authors.initials: F
authors.lastname: Gruber
authors.affiliation: Department of Dermatology, Medical University of Vienna, Anna Spiegel Gebäude E6 Lab5, 1090 Vienna, Austria.
authors.forename: Cayo Mecking
authors.initials: CM
authors.lastname: Ornelas
authors.affiliation: Department of Dermatology, Medical University of Vienna, Anna Spiegel Gebäude E6 Lab5, 1090 Vienna, Austria.
authors.forename: Susanne
authors.initials: S
authors.lastname: Karner
authors.affiliation: Department of Dermatology, Medical University of Vienna, Anna Spiegel Gebäude E6 Lab5, 1090 Vienna, Austria; Christian Doppler Laboratory for Biotechnology of Skin Aging, Vienna, Austria.
authors.forename: Marie-Sophie
authors.initials: MS
authors.lastname: Narzt
authors.affiliation: Department of Dermatology, Medical University of Vienna, Anna Spiegel Gebäude E6 Lab5, 1090 Vienna, Austria; Christian Doppler Laboratory for Biotechnology of Skin Aging, Vienna, Austria.
authors.forename: Ionela Mariana
authors.initials: IM
authors.lastname: Nagelreiter
authors.affiliation: Department of Dermatology, Medical University of Vienna, Anna Spiegel Gebäude E6 Lab5, 1090 Vienna, Austria.
authors.forename: Maria
authors.initials: M
authors.lastname: Gschwandtner
authors.affiliation: Department of Vascular Biology, Medical University of Vienna, 1090 Vienna, Austria.
authors.forename: Valery
authors.initials: V
authors.lastname: Bochkov
authors.affiliation: Department of Dermatology, Medical University of Vienna, Anna Spiegel Gebäude E6 Lab5, 1090 Vienna, Austria.
authors.forename: Erwin
authors.initials: E
authors.lastname: Tschachler
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002851:Chromatography, High Pressure Liquid; D004281:Docosahexaenoic Acids / Q000494:pharmacology*; D005347:Fibroblasts / Q000187:drug effects* / Q000378:metabolism / Q000528:radiation effects*; D006801:Humans; D007124:Immunoenzyme Techniques; D007249:Inflammation / Q000378:metabolism*; D015227:Lipid Peroxidation / Q000187:drug effects / Q000502:physiology / Q000528:radiation effects; D051379:Mice; D051267:NF-E2-Related Factor 2 / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012867:Skin / Q000187:drug effects / Q000378:metabolism / Q000528:radiation effects; D053719:Tandem Mass Spectrometry; D014466:Ultraviolet Rays / Q000009:adverse effects
chemical_list: D051267:NF-E2-Related Factor 2; D004281:Docosahexaenoic Acids