ANTIOXIDANT ACTIVITY OF EGT 25 mRNA expression. Thus, we examined the effect of EGT on MMP-1 mRNA expression in cultured normal human fibroblasts exposed to UVA. MMP-1 mRNA in human fibroblasts was elevated 1.25-fold at 24 h post UV A irradiation. EGT reduced MMP-1 mRNA expression levels in a dose-dependent manner (Figure 6). The results indicated that EGT down-regulated MMP-1 mRNA expression of fibroblasts induced by UVA irradiation. DISCUSSION EGT has structural similarities to histidine, urocanic acid, and carnitine, none of which are substantial antioxidants. EGT has strong and unusual antioxidant activity due to its sulfur in thione form, which converts to the sulfhydryl form to scavenge oxygen radicals. A recent study showed the protective abilities of EGT on cell damage induced by H202 and ONOO- (26). In addition, it suppressed NF-KB activation stimulated by H20 2 (27). Therefore, we focused on the antioxidative effects of EGT against •0 2 - and 1 0 2 , and examined whether it reduced the markers of skin aging. We show here that EGT scavenged •0 2 - and 1 0 2 and inhibited the lipid peroxidation initiated by these ROS. UV-irradiated fibroblasts secrete several proteins, including TNF-a, which is a pro­ inflammatory cytokine, and MMP-1, which degrades the collagen matrix. EGT inhib­ ited DVB-induced up-regulation of TNF-a gene expression and suppressed MMP-1 gene transcription in human dermal fibroblasts exposed to UV A. These data suggest that ROS can initiate these changes in gene expression. Cutaneous damage induced by UV irradiation may lead to premature skin aging, called photoaging (4). The relationship between wrinkle formation and dermal matrix alter­ ation has been actively studied by many researchers. It has been proposed that UV irradiation causes photoaging in skin by induction of an imbalance of dermal matrix metabolism, including reduction of collagen synthesis and enhancement of MMP-1 production. Recently, Chung et al. (28) proposed a theory on the dermal alterations in MMP-1 G3PDH EGT (mg/ml) UVA (J/cm2) 0 0 0 0.5 1.0 20 20 20 Figure 6. EGT-suppressed MMP-1 up-regulation by UVA irradiation. Human fibroblasts were exposed to UVA at a dose of 20 J/cm2 in the presence of various concentrations of EGT in HBS. Total RNA was extracted at 6 h after UVA irradiation. The expression of MMP-1 was evaluated by RT-PCR.
26 JOURNAL OF COSMETIC SCIENCE which MAP kinase signal transduction pathways shifted from extracellular regulated kinase-1 and -2 (ERK 1/2) to stress-activated protein kinase (SAPK), c-Jun amino­ terminal kinase, and p38 MAP kinase. In addition, UVA radiation in human skin fibroblasts induced p38 and JNK activity transiently but not extracellular signal­ regulated kinases (ERK) (29). The alteration of the signal transduction leads to wrinkle formation through the reduction of type I collagen synthesis and the increase in MMP production. Our data support the concept that these biological changes are caused in part by oxygen radicals, including hydrogen peroxide (H 2 0) and 1 0 2 (30,31). Taken together, EGT showed excellent scavenging ability against ROS and suppression of TNF-a and MMP-1 expression. It is well elucidated that ROS, TNF-a, and MMP-1 play important roles in UV-induced skin aging, particularly in wrinkle formation. Therefore, our findings may suggest that EGT prevents the progression of photoaging in skin. REFERENCES (1) 0. A. Bianciotto, L.B. Pinedo, N. A. San Roman, A. Y. Blessio, and M. B. Collantes, The effect of natural UV-B radiation on a perennial Salicornia salt-marsh in Bahia San Sebastian, Tierra del Fuego, Argentina: A 3-year field study.]. Photochem. Photobiol. B., 70, 177-185 (2003). (2) H. Alpermann and H. G. Vogel, Effect of repeated ultraviolet irradiation on skin of hairless mice, Arch. Dermatol. Res., 262, 15-25 (1978). (3) K. J. Johnston, A. I. Oikarinen, N. J. Lowe, J. G. Clark, and J. Uitto, Ultraviolet radiation-induced connective tissue changes in the skin of hairless mice,]. Invest. Dermatol., 82, 587-590 (1984). (4) L. H. Kligman, F. J. Akin, and A. M. Kligman, The contributions of UV A and UVB to connective tissue damage in hairless mice,]. Invest. Dermatol., 84, 272-276 (1985). (5) A. Oikarinen, J. Karvonen, J. Uitto, and M. Hannuksela, Connective tissue alterations in skin exposed to natural and therapeutic UV-radiation, Photodermatol., 2, 15-26 (1985). (6) I. Koshiishi, E. Horikoshi, H. Mitani, and T. Imanari, Quantitative alterations of hyaluronan and dermatan sulfate in the hairless mouse dorsal skin exposed to chronic UV irradiation, Biochirn. Biophys. Acta., 1428, 327-333 (1999). (7) Y. Nishimori, C. Edwards, A. Pearse, K. Matsumoto, M. Kawai, and R. Marks, Degenerative alter­ ations of dermal collagen fiber bundles in photodamaged human skin and UV-irradiated hairless mouse skin: Possible effect on decreasing skin mechanical properties and appearance of wrinkles,]. Invest. Dermatol., 117, 1458-1463 (2001). (8) H. Talwar, C. Griffiths, G. Fisher, T. Hamilton, and J. Voorhees, Reduction type I and type III procollagens in photodamaged adult human skin,]. Invest. Dermatol., 105, 285-290 (1995). (9) G. Fisher, Z. Wang, S. Datta, J. Varani, S. Kang, and J. Voorhees, Pathophysiology of premature skin aging induced by ultraviolet light, N. Engl.]. Med., 337, 1419-1428 0997). (10) V. M. Kahari and U. Saarialho-Kere, Matrix metalloproteinases in skin, Exp. Dennatol., 6, 199-213 (1997). (11) L. Rittie and G. J. Fisher, UV-light-induced signal cascades and skin aging, Aging Res. Rev., 1, 705-720 (2002). (12) C. Chadwick, C. Patten, 0. Nikaido, T. Matsunaga, C. P. Roby, and A. Young, The detection of cyclobutane thymine dimers, (6-4) photolesions and the Dewar photoisomers in sections of UV­ irradiated human skin using specific antibodies, and the demonstration of depth penetration effects, ]. Photochem. Photobiol. B, 28, 163-170 (1995). (13) A. Oxholm, P. Oxhold, B. Staberg, and K. Bendtzen, Immunohistological detection of interleukin I-like molecules and tumor necrosis factor in human epidermis before and after DVB-irradiation in vivo, Br.]. Dennatol., 118, 369-376 (1988). (14) H. Masaki and H. Sakurai, Increased generation of hydrogen peroxide possibly from mitochondrial respiratory chain after UVB irradiation ofmurine fibroblasts.]. Dennatol. Sci., 14, 207-216 (1997). (15) R. M. Tyrrell and M. Pidoux, Singlet oxygen involvement in the inactivation of cultured human
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