Abstract
The harmful effects of ultraviolet (UV) exposure on the skin are associated with the generation of reactive oxygen species (ROS) such as superoxide anion radical (•O2-), hydrogen peroxide (H2O2), hydroxyl radical (•OH), and singlet oxygen (1O2) as well as with lipid peroxides and their radicals (LOOH and LOO•). To give direct proof that such ROS are generated in UV-exposed skin, we proposed the in vivo detection and imaging method in which both a sensitive and specific chemiluminescence (CL) probe, such as CLA, and an ultralow-light imaging apparatus with a CCD camera were used. With this method we found that •O2- is formed intrinsically and that 1O2 and •O2- are generated in the UVA-exposed skin of mice. In addition, we indicated that antioxidative ability against ROS in the skin of hairless rats decreased as age increased. Using these findings, we demonstrated the protective abilities of sodium ascorbate, caffeic acid, essential aroma oils, and zinc(ii) ion and its complexes, which we administered to mice both topically and orally. We present a review for the current state of our research proposing the sensitive CL method as a useful in vivo tool in photobiological research for the detection of oxidative stress as well as for the evaluation of antioxidative agents to the skin.
Similar content being viewed by others
References
A. M. Kligman, Early destructive effect of sunlight on human skin, J. Am. Med. Assoc., 1969, 210, 2377–2380.
A. Oikarinen, J. Karvonen, J. Uitto and M. Hannuksela, Connective tissue alterations in skin exposed to natural and therapeutic UV-radiation, Photodermatology, 1985, 2, 15–26.
N. Bech-Thomsen and H. C. Wulf, Carcinogenic and melanogenic effects of a filtered metal halide UVA source and a tubular fluorescent UVA tanning source with or without additional solar-simulated UV radiation in hairless mice, Photochem. Photobiol., 1995, 62, 773–779.
J. Nishi, R. Ogura, M. Sugimoto, T. Hidaka and M. Kohno, Involvement of active oxygen in lipid peroxide radical reaction of epidermal homogenate following ultraviolet light exposure, J. Invest. Dermatol., 1991, 97, 115–119.
H. Masaki, T. Atsumi and H. Sakurai, Detection of hydrogen peroxide and hydroxyl radicals in murine skin fibroblasts under UVB irradiation, Biochem. Biophys. Res. Commun., 1995, 206, 474–479.
H. Masaki and H. Sakurai, Increased generation of hydrogen peroxide possibly from mitochondrial respiratory chain after UVB irradiation of murine fibroblasts, J. Dermatol. Sci., 1997, 14, 207–216.
P. Evelson, C. P. Ordonez, S. Llesuy and A. J. Boveris, Oxidative stress and in vivo chemiluminescence in mouse skin exposed to UVA radiation, Photochem. Photobiol., 1997, B 38, 215–219.
G. Sauermann, W. P. Mei, U. Hoppe and F. Stab, Ultraweak photon emission of human skin in vivo: influence of topically applied antioxidants on human skin, Methods Enzymol., 1999, 300, 419–428.
R. Gillies, G. Zonios, R. R. Anderson and N. Kollias, Fluorescence excitation spectroscopy provides information about human skin in vivo, J. Invest. Dermatol., 2000, 115, 704–707.
J. Wang, D. Xing, Y. He and X. Hu, Experimental study on photodynamic diagnosis of cancer mediated by chemiluminescence probe, FEBS Lett., 2002, 523, 128–132.
H. Ou-Yang, G. Stamatas, C. Saliou and N. Kollias, A chemiluminescence study of UVA-induced oxidative stress in human skin in vivo, J. Invest. Dermatol., 2004, 122, 1020–1029.
G. He, A. Samouilov, P. Kuppusamy and J. L. Zweir, In vivo EPR imaging of the distribution and metabolism of nitroxide radicals in human skin, J. Mag. Reson., 2001, 148, 155–164.
T. Herrling, J. Fuchs, J. Rehberg and N. Groth, UV-induced free radicals in the skin detected by ESR spectroscopy and imaging using nitroxides, Free Rad. Biol. Med., 2003, 35, 59–67.
G. He, V. K. Kutala, P. Kuppusamy and J. L. Zweir, In vivo measurement and mapping of skin redox stress induced by ultraviolet light exposure, Free Rad. Biol. Med., 2004, 36, 665–672.
H. Yasui and H. Sakurai, Chemiluminescent detection and imaging of reactive oxygen species in live mouse skin exposed to UVA, Biochem. Biophys. Res. Commun., 2000, 269, 131–136.
H. Yasui and H. Sakurai, Age-dependent generation of reactive oxygen species in the skin of live hairless rats exposed to UVA light, Exp. Dermatol., 2003, 12, 655–661.
M. Ozaki, S. Kawashima, T. Yamashita, T. Hirase, M. Namiki, N. Inoue, K. Hirata, H. Yasui, H. Sakurai, Y. Yoshida, M. Masada and M. Yokoyama, Overexpression of endothelial nitric oxide synthase accelerates atherosclerotic lesion formation in apoE-deficient mice, J. Clin. Invest., 2002, 110, 331–340.
H. Sakurai, H. Yasui, K. Mishina and H. Nishimura, Protective effect of zinc ion on ultraviolet light-induced skin damage: studies on imaging of ROS by chemiluminescence method, Biomed. Res. Trace Elements, 2003, 14, 27–21.
H. Yasui, A. Ikeda, M. Shigemoto and H. Sakurai, Dermatological suppressive effect of lavender oil against UVA-induced reactive oxygen species (ROS) in the skin of live animals: possibility of lavender oil for both protection of ultraviolet light-induced dermal injury and suppression of skin-aging, Aroma Res., 2004, 18, 129–135, (in Japanese).
C. E. Hooper, R. E. Ansorge, H. M. Browne and P. Tomkins, CCD imaging of luciferase gene expression in single mammalian cells, J. Biolumin. Chemilumin., 1990, 5, 123–130.
J. C. Nicolas, Applications of low-light imaging to life sciences, J. Biolumin. Chemilumin., 1994, 9, 139–144.
A. Roda, P. Pasini, M. Musiani, S. Girotti, M. Baraldini, G. Carrea and A. Suozzi, Chemiluminescent low-light imaging of biospecific reactions on macro- and microsamples using a videocamera-based luminograph, Anal. Chem., 1996, 68, 1073–1080.
D. Stawinska and J. Stawinski, Chemiluminescence of cereal products III. Two-dimensional photocount imaging of chemiluminescence, J. Biolumin. Chemilumin., 1998, 13, 21–24.
G. Rotilio, R. C. Bray and E. M. Fielden, A pulse radiolysis study of superoxide dismutase, Biochem. Biophys. Acta, 1972, 268, 605–609.
D. Bellus, Physical quenchers of singlet molecular oxygen, Adv. Photochem., 1979, 11, 105–205.
D. Gao, M. Kakuma, S. Oka, K. Sugino and H. Sakurai, Reaction of ß-alkanin (Shikonin) with reactive oxygen species: detection of ß-alkanin free radicals, Bioorg. Med. Chem., 2000, 8, 2561–2569.
H. Yasui and H. Sakurai, MCLA-dependent chemiluminence method for detection of peroxyl radical, using t-BuOO• as a model species, submitting.
P. Hasty, J. Campisi, J. Hoeijmakers, H. van Steeg and J. Vijg, Aging and genome maintenance: lessons from the mouse?, Science, 2003, 299, 1355–1359.
Y. Yamada, H. Yasui and H. Sakurai, Suppressive effect of caffeic acid and its derivatives on the generation of UVA-induced reactive oxygen species in the skin of live mice, submitting.
A. S. Prasad, Clinical, biochemical and nutritional spectrum of zinc deficiency in human subjects: an update, Nutr. Rev., 1983, 41, 197–208.
C. T. Dillon, T. W. Hambley, B. J. Kennedy, P. A. Lay, Q. Zhou, N. M. Davies, J. R. Biffin and H. L. Regtop, Gastrointestinal toxicity, anti-inflammatory activity and superoxide dismutase activity of copper and zinc complexes of anti-inflammatory drug indomethacin, Chem. Res. Toxicol., 2003, 16, 28–37.
B. A. Igor, B. S. Tatjana, P. C. Zinaida, E. A. Natalia and G. K. Ludmila, Study of antioxidant properties of metal aspartates, Analyst, 1995, 120, 859–862.
M. S. Ågren, Influence of two vehicles for zinc oxide on zinc absorption through intact skin and wounds, Acta. Derm. Venereol., 1991, 71, 153–156.
H. Sakurai, H. Nishimura, K. Mishina and H. Yasui, Oral zinc supplementation suppresses the formation of reactive oxygen species in the skin of live animals under ultra-violet light exposure, 4th lnt. Symp. Trace Elem. Humans: New Persp., Proceedings Book Part 1, ed. S. Ermidou-Pollet and S. Pollet, Entypossis, Athens, Greece, 2003, pp. 590–601.
Y. Yoshikawa, E. Ueda, K. Kawabe, H. Miyake, T. Takino, H. Sakurai and Y. Kojima, Development of new insulinomimetic zinc(ii) picolinate complexes with a Zn(N2O2) coordination mode: structure characterization, in vitro and in vivo studies, J. Biol. Inorg. Chem., 2002, 7, 68–73.
H. Masaki, Y. Okano and H. Sakurai, Induction of metallothioneins in HaCaT cells treated with ZnCl2 and Zn(ii) complexes, manuscript in preparation.
C. S. Sander, H. Chang, S. Salzmann, C. S. L. Müller, S. Ekanayake-Mudiyanselage, P. Elsner and J. J. Thiele, Photoaging in associated with protein oxidation in human skin in vivo, J. Invest. Dermatol., 2002, 118, 618–625.
L. Hellemans, H. Corstjens, A. Neven, L. Declercq and D. Maes, Antioxidant enzyme activity in human stratum corneum shows seasonal variation with an age-dependent recovery, J. Invest. Dermatol., 2003, 120, 434–439.
Author information
Authors and Affiliations
Corresponding author
Additional information
Presented at the 14th International Congress on Photobiology, at Jungmoon, Jeju Island, South Korea, 10th-15th June 2004.
Rights and permissions
About this article
Cite this article
Sakurai, H., Yasui, H., Yamada, Y. et al. Detection of reactive oxygen species in the skin of live mice and rats exposed to UVA light: a research review on chemiluminescence and trials for UVA protection. Photochem Photobiol Sci 4, 715–720 (2005). https://doi.org/10.1039/b417319h
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1039/b417319h