Protection by Food-derived Antioxidants from UV-A1–Induced Photodamage, Measured Using Living Skin Equivalents¶

Abstract In a study of biomarkers of ultraviolet-A1 radiation (UV-A1)-induced skin damage, living skin equivalent cultures (LSE) were treated with the antioxidants hesperetin and quercetin-3-glucoside and irradiated with 25 or 50 J/cm2 UV-A1. Changes in the following biomarkers were measured; Interleukin 1-alpha (IL-1α), Heme Oxygenase-1 (HO-1), TdT-mediated dUTP nick end labeling (TUNEL) and 8-hydroxy-2′-deoxyguanosine (8-OHdG). IL-1α and HO-1 were analyzed by real-time PCR, Western blot, enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry. TUNEL and 8-OHdG were determined by (immuno)histochemical techniques. Sections were stained with hematoxylin and eosin (H&E). UV-A1 induced keratinocyte and fibroblast vacuolation and nuclear pyknosis, intense TUNEL staining of fibroblasts and increased staining of cells and nuclei for 8-OHdG. Lesser or marginal increases in intensity followed staining for HO-1 and IL-1α. The IL-1α increase was confirmed by ELISA assays of the medium supernatants. Hesperetin and quercetin-3-glucoside reduced changes in H&E, 8-OHdG, TUNEL and IL-1α. Quercetin-3-glucoside reduced the amount of IL-1α in LSE media. These observations support the use of the selected biomarkers to monitor UV-A1 damage and provide evidence that dietary ingredients could reduce ultraviolet-A radiation-induced damage.

[1]  V. Reeve,et al.  Immunoprotective UVA (320-400 nm) irradiation upregulates heme oxygenase-1 in the dermis and epidermis of hairless mouse skin. , 2004, The Journal of investigative dermatology.

[2]  A. Svobodová,et al.  Natural phenolics in the prevention of UV-induced skin damage. A review. , 2003, Biomedical papers of the Medical Faculty of the University Palacky, Olomouc, Czechoslovakia.

[3]  R. Tyrrell,et al.  Hesperetin Glucuronide, a Photoprotective Agent Arising from Flavonoid Metabolism in Human Skin Fibroblasts¶ , 2003, Photochemistry and photobiology.

[4]  J. Youn,et al.  Changes of comedonal cytokines and sebum secretion after UV irradiation in acne patients. , 2002, European journal of dermatology : EJD.

[5]  H. Hendriks,et al.  Nutritional skin care: health effects of micronutrients and fatty acids. , 2001, The American journal of clinical nutrition.

[6]  S. Werner,et al.  Haem oxygenase-1: a novel player in cutaneous wound repair and psoriasis? , 2001, The Biochemical journal.

[7]  T. Gambichler,et al.  UV Transmission Measurements of Small Skin Specimens with Special Quartz Cuvettes , 2000, Dermatology.

[8]  S. Ryter,et al.  Cellular studies with UVA radiation: A role for iron , 2000 .

[9]  C. Scaletta,et al.  Expression of DNA damage and stress proteins by UVA irradiation of human skin in vivo , 2000 .

[10]  K. Herbert,et al.  Immunochemical quantitation of UV-induced oxidative and dimeric DNA damage to human keratinocytes , 2000, Free radical research.

[11]  H. Bonkovsky,et al.  Heme oxygenase: recent advances in understanding its regulation and role. , 1999, Proceedings of the Association of American Physicians.

[12]  S. Kondo,et al.  The roles of keratinocyte-derived cytokines in the epidermis and their possible responses to UVA-irradiation. , 1999, The journal of investigative dermatology. Symposium proceedings.

[13]  R. Tyrrell,et al.  Heme oxygenase induction mediates the photoimmunoprotective activity of UVA radiation in the mouse. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[14]  J. Mergny,et al.  UV-A-induced decrease in nuclear factor-kappaB activity in human keratinocytes. , 1999, The Biochemical journal.

[15]  C. Potten,et al.  Human melanocytes and keratinocytes exposed to UVB or UVA in vivo show comparable levels of thymine dimers. , 1998, The Journal of investigative dermatology.

[16]  R. Tyrrell,et al.  Modulation of the UVA activation of haem oxygenase, collagenase and cyclooxygenase gene expression by epigallocatechin in human skin cells , 1998, FEBS letters.

[17]  F. Bernerd,et al.  UVA exposure of human skin reconstructed in vitro induces apoptosis of dermal fibroblasts: subsequent connective tissue repair and implications in photoaging , 1998, Cell Death and Differentiation.

[18]  T. Nagaraja,et al.  Intracellular acidification induced by passive and active transport of ammonium ions in astrocytes. , 1998, American journal of physiology. Cell physiology.

[19]  J. Hoidal,et al.  Effect of tumor necrosis factor-alpha and interleukin-1 alpha on heme oxygenase-1 expression in human endothelial cells. , 1998 .

[20]  J. Hoidal,et al.  Effect of tumor necrosis factor-α and interleukin-1α on heme oxygenase-1 expression in human endothelial cells. , 1998, American journal of physiology. Heart and circulatory physiology.

[21]  R. Tyrrell,et al.  Development of Refractoriness of Induced Human Heme Oxygenase‐1 Gene Expression to Reinduction by UVA Irradiation and Hemin , 1997, Photochemistry and photobiology.

[22]  S. R. Feldman,et al.  Epidermal stratification reduces the effects of UVB (but not UVA) on keratinocyte cytokine production and cytotoxicity , 1997, Photodermatology, photoimmunology & photomedicine.

[23]  S. Grether-Beck,et al.  Induction of Proinflammatory Cytokines in Human Epidermoid Carcinoma Cells by in vitro Ultraviolet A1 Irradiation , 1997, Photochemistry and photobiology.

[24]  F. Bernerd,et al.  Successive alteration and recovery of epidermal differentiation and morphogenesis after specific UVB-damages in skin reconstructed in vitro. , 1997, Developmental biology.

[25]  J. Eberle,et al.  L-ascorbic acid inhibits UVA-induced lipid peroxidation and secretion of IL-1alpha and IL-6 in cultured human keratinocytes in vitro. , 1997, The Journal of investigative dermatology.

[26]  O. Damour,et al.  Use of dermal equivalent and skin equivalent models for identifying phototoxic compounds in vitro , 1997, Photodermatology, photoimmunology & photomedicine.

[27]  S. Toyokuni,et al.  8-hydroxy-2'-deoxyguanosine is increased in epidermal cells of hairless mice after chronic ultraviolet B exposure. , 1996, The Journal of investigative dermatology.

[28]  J. Mergny,et al.  Ultraviolet‐A induces activation of AP‐1 in cultured human keratinocytes , 1996, FEBS letters.

[29]  R. Tyrrell,et al.  Activation of mammalian gene expression by the UV component of sunlight – from models to reality , 1996, BioEssays : news and reviews in molecular, cellular and developmental biology.

[30]  L. Applegate,et al.  Oxidative defense in cultured human skin fibroblasts and keratinocytes from sun‐exposed and non‐exposed skin , 1995, Photodermatology, photoimmunology & photomedicine.

[31]  J. H. Chung,et al.  Effect of ultraviolet A on IL-1 production by ultraviolet B in cultured human keratinocytes. , 1995, Journal of dermatological science.

[32]  R. Tyrrell,et al.  TWO GENES CONTRIBUTE TO DIFFERENT EXTENTS TO THE HEME OXYGENASE ENZYME ACTIVITY MEASURED IN CULTURED HUMAN SKIN FIBROBLASTS AND KERATINOCYTES: IMPLICATIONS FOR PROTECTION AGAINST OXIDANT STRESS , 1995, Photochemistry and photobiology.

[33]  R. Tyrrell,et al.  Heme oxygenase 1 mediates an adaptive response to oxidative stress in human skin fibroblasts. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[34]  R. Tyrrell,et al.  Endogenous glutathione levels modulate both constitutive and UVA radiation/hydrogen peroxide inducible expression of the human heme oxygenase gene. , 1992, Carcinogenesis.

[35]  M. Darmon,et al.  Epidermal morphogenesis and induction of the 67 kD keratin polypeptide by culture of human keratinocytes at the liquid-air interface. , 1985, Experimental cell research.

[36]  C. Scaletta,et al.  Photoprotective potential of lycopene, beta-carotene, vitamin E, vitamin C and carnosic acid in UVA-irradiated human skin fibroblasts. , 2002, Free radical biology & medicine.

[37]  R. Tyrrell.  Redox regulation and oxidant activation of heme oxygenase-1. , 1999, Free radical research.

[38]  R. Patel,et al.  Direct detection of 8-oxodeoxyguanosine and 8-oxoguanine by avidin and its analogues. , 1998, Analytical biochemistry.