title: Skin resistance to oxidative stress induced by resveratrol: from Nrf2 activation to GSH biosynthesis.
abstract: Skin is particularly exposed to oxidative stress, either from environmental insults such as sunlight or pollution or as a consequence of specific impairments in antioxidant status resulting from pathologies or aging. Traditionally, antioxidant products are exogenously provided to neutralize pro-oxidant species. However, another approach based on stimulation of endogenous antioxidant defense pathways is more original. Resveratrol (RSV) was reported to display such a behavior in various tissues, but data about the mechanisms of action in skin are scarce. We show here that, in primary culture of normal human keratinocytes (NHKs) or in full-thickness reconstructed human skin, RSV activated the Nrf2 pathway at nontoxic doses, from 20 µM up to 100µM. Among the Nrf2 downstream genes, glutamylcysteinyl ligase and glutathione peroxidase-2 were induced at the mRNA and protein levels. In parallel, a significant increase in glutathione content, assessed by LC/MS analysis, was observed in both models. Nrf2 gene silencing experiments performed in NHKs confirmed that Nrf2 was involved in RSV-induced modulation of cellular antioxidant status, in part by increasing cellular glutathione content. Finally, improvement of endogenous defenses induced in RSV-pretreated reconstructed skin ensured protection against the toxic oxidative effects of cumene hydroperoxide (CHP). In fact after RSV pretreatment, in response to CHP stress, glutathione content did not decrease as in unprotected samples. Cellular alterations at the dermal-epidermal junction were clearly prevented. Together, these complementary experiments demonstrated the beneficial effects of RSV on skin, beyond its direct antioxidant properties, by upregulation of a cutaneous endogenous antioxidant pathway.
authors.affiliation: L'Oréal Research and Innovation, 93600 Aulnay-sous-Bois, France. Electronic address: jsoeur@rd.loreal.com.
authors.forename: J
authors.initials: J
authors.lastname: Soeur
authors.affiliation: L'Oréal Research and Innovation, 93600 Aulnay-sous-Bois, France.
authors.forename: J
authors.initials: J
authors.lastname: Eilstein
authors.affiliation: L'Oréal Research and Innovation, 93600 Aulnay-sous-Bois, France.
authors.forename: G
authors.initials: G
authors.lastname: Léreaux
authors.affiliation: L'Oréal Research and Innovation, 93600 Aulnay-sous-Bois, France.
authors.forename: C
authors.initials: C
authors.lastname: Jones
authors.affiliation: L'Oréal Research and Innovation, 93600 Aulnay-sous-Bois, France.
authors.forename: L
authors.initials: L
authors.lastname: Marrot
mesh_terms: D000975:Antioxidants / Q000494:pharmacology*; D015153:Blotting, Western; D002478:Cells, Cultured; D002853:Chromatography, Liquid; D005978:Glutathione / Q000378:metabolism*; D006801:Humans; D015603:Keratinocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D051267:NF-E2-Related Factor 2 / Q000235:genetics / Q000378:metabolism*; D018384:Oxidative Stress; D021381:Protein Transport; D012333:RNA, Messenger / Q000235:genetics; D017382:Reactive Oxygen Species / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D000077185:Resveratrol; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012869:Skin Absorption / Q000187:drug effects*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013267:Stilbenes / Q000494:pharmacology*
chemical_list: D000975:Antioxidants; D051267:NF-E2-Related Factor 2; D012333:RNA, Messenger; D017382:Reactive Oxygen Species; D013267:Stilbenes; D005978:Glutathione; D000077185:Resveratrol
keywords: Free radicals; Glutathione; Normal human keratinocyte; Nrf2; Oxidative stress; Reconstructed human skin model; Resveratrol