Abstract
Ultraviolet (UV) radiation is the major environmental factor affecting functions of the skin. Compounds rich in polyphenols, such as Thymus vulgaris leaf extract and thymol, have been proposed for the prevention of UV-induced skin damage. We compared the acute effects induced by UVA and UVB rays on epidermal morphology and proliferation, cytotoxicity, and genotoxicity. Normal human skin explants were obtained from young healthy women (n = 7) after informed consent and cultured at the air-liquid interface overnight. After 24 h, the samples were divided in 2 groups: the former exposed to UVA (16 or 24 J/cm2) and the latter irradiated with UVB (0.24 or 0.72 J/cm2). One hour after the end of irradiation, supernatants were collected for evaluation of the lactate dehydrogenase activity. Twenty-four hours after UVB exposure, biopsies were processed for light and transmission electron microscopy analysis, proliferation, cytotoxicity, and genotoxicity. UVB and UVA rays induced early inhibition of cell proliferation and DNA damage compared to controls. In particular, UVB rays were always more cytotoxic and genotoxic than UVA ones. For this reason, we evaluated the effect of either T. vulgaris L. extract (1.82 µg/ml) or thymol (1 µg/ml) on all samples treated for 1 h before UVB irradiation. While Thymus had a protective action for all of the endpoints evaluated, the action of the extract was less pronounced on epidermal proliferation and morphological features. The results presented in this study could be the basis for investigating the mechanism of thymol and T. vulgaris L. extract against the damage induced by UV radiation.
References
1.
Aeschbach, R., J. Löliger, B.C. Scott, A. Murcia, J. Butler, B. Halliwell, O.I. Aruoma (1994) Antioxidant actions of thymol, carvacrol, 6-gingerol, zingerone and hydroxytyrosol. Food Chem Toxicol 32: 31-36.
2.
Afaq, F., S.K. Katiyar (2011) Polyphenols: skin photoprotection and inhibition of photocarcinogenesis. Mini Rev Med Chem 11: 1200-1215.
3.
Alghamdi, K.M., H. Khurram, A. Taïeb (2012) Survey of dermatologists' phototherapy practices for vitiligo. Indian J Dermatol Venereol Leprol 78: 74-81.
4.
Aoki-Yoshida, A., R. Aoki, Y. Takayama (2013) Protective effect of pyruvate against UVB-induced damage in HaCat human keratinocytes. J Biosci Bioeng 115: 442-448.
5.
Archana, P.R., B. Nageshwar Rao, M. Ballal, B.S. Satish Rao (2009) Thymol, a naturally occurring monocyclic dietary phenolic compound protects Chinese hamster lung fibroblasts from radiation-induced cytotoxicity. Mutat Res 680: 70-77.
6.
Arcila-Lozano, C.C., G. Loarca-Piña, S. Lecona-Uribe, E. González de Mejía (2004) Oregano: properties, composition and biological activity (in Spanish). Arch Latinoam Nutr 54: 100-111.
7.
Auger, F.A., F. Berthod, V. Moulin, R. Pouliot, L. Germain (2004) Tissue-engineered skin substitutes: from in vitro constructs to in vivo applications. Biotechnol Appl Biochem 39: 263-275.
8.
Bedoni, M., C. Sforza, C. Dolci, E. Donetti (2007) Proliferation and differentiation biomarkers in normal human breast skin organotypic cultures. J Dermatol Sci 46: 139-142.
9.
Bernard, F., D. Asselineau (1997) Successive alteration and recovery of epidermal differentiation and morphogenesis after specific UVB-damages in skin reconstructed in vitro. Dev Biol 183: 123-138.
10.
Bozin, B., N. Mimica-Dukic, N. Simin, G. Anackov (2006) Characterization of the volatile composition of essential oils of some Lamiaceae spices and the antimicrobial and antioxidant activities of the entire oils. J Agric Food Chem 54: 1822-1828.
11.
Braga, P.C., M. Dal Sasso, M. Culici, L. Galastri, M.T. Marceca, E.E. Guffanti (2006) Antioxidant potential of thymol determined by chemiluminescence inhibition in human neutrophils and cell-free systems. Pharmacology 76: 61-68.
12.
Byrne, S.N., C. Beaugie, C. O'Sullivan, S. Leighton, G.M. Halliday (2011) The immune-modulating cytokine and endogenous alarmin interleukin-33 is upregulated in skin exposed to inflammatory UVB radiation. Am J Pathol 179: 211-222.
13.
Cadenas, E. (2000) Mitochondrial free radical generation, oxidative stress and aging. Free Radic Biol Med 29: 222-230.
14.
Cadet, J., T. Douki (2011) Oxidatively generated damage to DNA by UVA radiation in cells and human skin. J Invest Dermatol 131: 1005-1007.
15.
Calò, R., L. Marabini (2014) Protective effect of Vaccinum myrtillus extract against UVA- and UVB-induced damage in a human keratinocyte cell line (HaCaT cells). J Photochem Photobiol 132: 27-35.
16.
Calò, R., C.M. Visone, L. Marabini (2015) Thymol and Thymus vulgaris L. activity against UVA- and UVB-induced damage in NCTC 2544 cell line. Mutat Res Genet Toxicol Environ Mutagen 791: 30-37.
17.
Didry, N., L. Dubreuil, M. Pinkas (1994) Activity of thymol, carvacrol, cinnamaldehyde and eugenol on oral bacteria. Pharm Acta Helv 69: 25-28.
18.
Donetti, E., M. Bedoni, E. Boschini, A.A. Bertelli, C. Sforza, N. Gagliano (2005) Early epidermal response after a single dose of gamma-rays in organotypic culture of human breast skin. Br J Dermatol 153: 881-886.
19.
Donetti, E., L. Cornaghi, A. Gualerzi, F.W. Baruffaldi Preis, F. Prignano (2014) An innovative three-dimensional model of normal human skin to study the proinflammatory psoriatic effects of tumor necrosis factor-alpha and interleukin-17. Cytokine 68: 1-8.
20.
Fernández-García, E. (2014) Skin protection against UV light by dietary antioxidants. Food Funct 5: 1994-2003.
21.
Fischer, T.W., K. Kleszczyński, L.H. Hardkop, N. Kruse, D. Zillikens (2013) Melatonin enhances antioxidative enzyme gene expression (CAT, GPx, SOD), prevents their UVR-induced depletion, and protects against the formation of DNA damage (8-hydroxy-2′-deoxyguanosine) in ex vivo human skin. J Pineal Res 54: 303-312.
22.
Fujisawa, S., Y. Kadoma (1992) Effect of phenolic compounds on the polymerization of methyl methacrylate. Dent Mater 8: 324-326.
23.
Grosso, C., A.C. Figueiredo, J. Burillo, A.M. Mainar, J.S. Urieta, J.G. Barroso, J.A. Coelho, A.M.F. Palavra (2010) Composition and antioxidant activity of Thymus vulgaris volatiles: comparison between supercritical fluid extraction and hydrodistillation. J Sep Sci 33: 2211-2218.
24.
Halliday, G.M., S.N. Byrne, D.L. Damian (2011) Ultraviolet A radiation: its role in immunosuppression and carcinogenesis. Semin Cutan Med Surg 30: 214-221.
25.
Hashemipour, H., H. Kermanshahi, A. Golian, T. Veldkamp (2013) Effect of thymol and carvacrol feed supplementation on performance, antioxidant enzyme activities, fatty acid composition, digestive enzyme activities, and immune response in broiler chickens. Poult Sci 92: 2059-2069.
26.
Horosová, K., D. Bujnáková, V. Kmet (2006) Effect of oregano essential oil on chicken lactobacilli and E. coli. Folia Microbiol 51: 278-280.
27.
Hudaib, M., E. Speroni, A.M. Di Pietra, V. Cavrini (2002) GC/MS evaluation of thyme (Thymus vulgaris L.) oil composition and variations during the vegetative cycle. J Pharm Biomed Anal 29: 691-700.
28.
Ichihashi, M., M. Ueda, A. Budiyanto, T. Bito, M. Oka, M. Fukunaga, K. Tsuru, T. Horikawa (2003) UV-induced skin damage. Toxicology 189: 21-39.
29.
Ikeahata, H., T. Ono (2011) The mechanisms of UV mutagenesis. J Radiat Res 52: 115-125.
30.
Johnson-Huang, L.M., M. Suarez-Farinas, M. Sullivan-Whalen, P. Gilleaudeau, J.G. Krueger, M.A. Lowes (2010) Effective narrow-band UVB radiation therapy suppresses the IL-23/ IL-17 axis in normalized psoriasis plaques. J Invest Dermatol 130: 2654-2663.
31.
Juzeniene, A., J. Moan (2012) Beneficial effects of UV radiation other than via vitamin D production. Dermatoendocrinol 4: 109-117.
32.
Katiyar, S.K. (2011) Green tea prevents non-melanoma skin cancer by enhancing DNA repair. Arch Biochem Biophys 508: 152-158.
33.
Khan, N., F. Afaq, M. Saleem, N. Ahmad, H. Mukhtar (2006) Targeting multiple signaling pathways by green tea polyphenol (-)-epigallocatechin-3-gallate. Cancer Res 66: 2000-2005.
34.
Kim, Y.A., B.T. Choi, Y.T. Lee, D.I. Park, S.H. Rhee, K.Y. Park, Y.H. Choi (2004) Resveratrol inhibits cell proliferation and induces apoptosis of human breast carcinoma MCF-7 cells. Oncol Rep 11: 441-446.
35.
Kim, Y.S., J.W. Hwang, S.H. Kang, E.H. Kim, Y.J. Jeon, J.H. Jeong, H.R. Kim, S.H. Moon, B.T. Jeon, P.J. Park (2014) Thymol from Thymus quinquecostatus Celak. protects against tert-butyl hydroperoxide-induced oxidative stress in Chang cells. J Nat Med 68: 154-162.
36.
Kirke, S.M., S. Lowder, J.J. Lloyd, B.L. Diffey, J.N. Matthews, P.M. Farr (2007) A randomized comparison of selective broadband UVB and narrowband UVB in the treatment of psoriasis. J Invest Dermatol 127: 1641-1646.
37.
Lambert, J.D., J. Hong, G.Y. Yang, J. Liao, C.S. Yang (2005) Inhibition of carcinogenesis by polyphenols: evidence from laboratory investigations. Am J Clin Nutr 81: 284S-291S.
38.
Mantena, S.K, S.M. Meeran, C.A. Elmets, S.K. Katiyar (2005) Orally administered green tea polyphenols prevent ultraviolet radiation-induced skin cancer in mice through activation of cytotoxic T cells and inhibition of angiogenesis in tumors. J Nutr 135: 2871-2877.
39.
Meeran, S.M., S. Akhtar, S.K. Katiyar (2009) Inhibition of UVB-induced skin tumor development by drinking green tea polyphenols is mediated through DNA repair and subsequent inhibition of inflammation. J Invest Dermatol 129: 1258-1270.
40.
Meeran, M.F., P.S. Prince (2012) Protective effects of thymol on altered plasma lipid peroxidation and nonenzymic antioxidants in isoproterenol-induced myocardial infarcted rats. J Biochem Mol Toxicol 26: 368-373.
41.
Miyamura, Y., S.G. Coelho, K. Schlenz, J. Batzer, C. Smuda, W. Choi, M. Brenner, T. Passeron, G. Zhang, L. Kolbe, R. Wolber, V.J. Hearing (2011) The deceptive nature of UVA tanning versus the modest protective effects of UVB tanning on human skin. Pigment Cell Melanoma Res 24: 136-147.
42.
Murapa, P., J. Dai, M. Chung, R.J. Mumper, J. D'Orazio (2012) Anthocyanin-rich fractions of blackberry extracts reduce UV-induced free radicals and oxidative damage in keratinocytes. Phytother Res 26: 106-112.
43.
Ocaña, A., G. Reglero (2012) Effects of thyme extract oils (from Thymus vulgaris, Thymus zygis, and Thymus hyemalis) on cytokine production and gene expression of oxLDL-stimulated THP-1-macrophages. J Obes 2012: 1-11.
44.
Piskin, G., U. Tursen, R.M. Sylva-Steenland, J.D. Bos, M.B. Teunissen (2004) Clinical improvement in chronic plaque-type psoriasis lesions after narrow-band UVB therapy is accompanied by a decrease in the expression of IFN-gamma inducers - IL-12, IL-18 and IL-23. Exp Dermatol 13: 764-772.
45.
Prignano, F., F. Arnaboldi, L. Cornaghi, F. Landoni, L. Tripo, F.W. Baruffaldi Preis, E. Donetti (2015) Tumor necrosis factor-alpha and interleukin-17 differently affects Langerhans cell distribution and activation in an innovative three-dimensional model of normal human skin. Eur J Cell Biol 94: 71-77.
46.
Rácz, E., E.P. Prens, D. Kurek, M. Kant, D. de Ridder, S. Mourits, E.M. Baerveldt, Z. Ozgur, W.F. van IJcken, J.D. Laman, F.J. Staal, L. van der Fits (2011) Effective treatment of psoriasis with narrow-band UVB phototherapy is linked to suppression of the IFN and Th17 pathways. J Invest Dermatol 131: 1547-1558.
47.
Rastogi, R.P., A. Richa Kumar, M.B. Tyagy, R.P. Sinha (2010) Molecular mechanisms of ultraviolet radiation-induced DNA damage and repair. J Nucleic Acids 2010: 592980.
48.
Reus, A.A., U. Mustafa, C.A.M. Krul (2012) The use of ex vivo human skin tissue for genotoxicity testing, Toxicol Appl Pharmacol 261: 154-163.
49.
Riella, K.R., R.R. Marinho, J.S. Santos, R.N. Pereira-Filho, J.C. Cardoso, R.L.C. Albuquerque-Junior, S.M. Thomazzi (2012) Anti-inflammatory and cicatrizing activities of thymol, a monoterpene of the essential oil from Lippia gracilis, in rodents. J Ethnopharmacol 143: 656-663.
50.
Salucci, S., S. Burattini, D. Curzi, F. Buontempo, A.M. Martelli, G. Zappia, E. Falcieri, M. Battistelli (2014) Antioxidants in the prevention of UVB-induced keratinocyte apoptosis. J Photochem Photobiol B 141: 1-9.
51.
Satooka, H., I. Kubo (2012) Effects of thymol on B16-F10 melanoma cells. J Agric Food Chem 60: 2746-2752.
52.
Silva, A.R., C. Seidl, A.S. Furusho, M.M. Boeno, G.C. Dieamant, A.M. Weffort-Santos (2013) In vitro evaluation of the efficacy of commercial green tea extracts in UV protection. Int J Cosmet Sci 35: 69-77.
53.
Sklar, L.R., F. Almutawa, H.W. Lim, I. Hamzavi (2013) Effects of ultraviolet radiation, visible light, and infrared radiation on erythema and pigmentation: a review. Photochem Photobiol Sci 12: 54-64.
54.
Suzuki, Y., H. Furuta (1988) Stimulation of guinea pig neutrophil superoxide anion-producing system with thymol. Inflammation 12: 575-584.
55.
ten Berge, O., S.G. van Velsen, B. Giovannone, C.A. Bruijnzeel-Koomen, E.F. Knol, K. Guikers, H. van Weelden (2011) Assessment of cyclobutane pyrimidine dimers by digital photography in human skin. J Immunol Methods 373: 240-246.
56.
Undeğer, U., A. Başaran, G.H. Degen, N. Başaran (2009) Antioxidant activities of major thyme ingredients and lack of (oxidative) DNA damage in V79 Chinese hamster lung fibroblast cells at low levels of carvacrol and thymol. Food Chem Toxicol 47: 2037-2043.
57.
Wacker, M., M.F. Holick (2013) Sunlight and vitamin D: a global perspective for health. Dermatoendocrinol 5: 51-108.
58.
Wang, L.S., G.D. Stoner (2008) Anthocyanins and their role in cancer prevention. Cancer Lett 269: 281-290.
59.
Wong, R., S. Geyer, W. Weninger, J.C. Guimberteau, J.K. Wong (2015) The dynamic anatomy and patterning of skin. Exp Dermatol 25: 92-98.
60.
Yang, C.S., X. Wang, G. Lu, S.C. Picinich (2009) Cancer prevention by tea: animal studies, molecular mechanisms and human relevance. Nat Rev Cancer 9: 429-439.
© 2016 S. Karger AG, Basel
2016
Copyright / Drug Dosage / Disclaimer
Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.
You do not currently have access to this content.
