22 JOURNAL OF COSMETIC SCIENCE 60.0 50.0 - E 40.0 0 30.0 C -E a 20.0 D.. ..J 10.0 0.0 0 Cl) I- G) G) G) I- 0 en ... C CJ C C C :c C ::z ... 0 w 'ii · m cu C z C 0 0 .... ... .c .... Cl) Cl) ... cu u .c � � Cl) .c (.) y 0 C. I ...J .... en E ...J C. I cu � .c ..J CJ ... D.. Cl) Cl) � (.) E cu -e I I z ca. 0 I CJ ..J en Figure 3. Scavenging effect of antioxidants against superoxide anion generated by alloxan. Lipid peroxides (LPO) were generated in liposomes by alloxan without addition (none). The level of LPO in samples with 20 µM antioxidants was measured after 60 min. 1 0 2 are shown in Table I. Rose bengal plus visible light was used as the source of 1 0 2 . The LPO level of control liposomes was 23.81 nmol/ml, and the exposure to 1 0 2 increased LPO to 91.84 nmol/ml. The addition of EGT reduced LPO to 26.53 nmol/ml, a 96% reduction (Table I). TNF-a EXPRESSION BY UVB IRRADIATION To examine the effects of EGT on DVB-induced TNF-a expression, we carried out a reporter assay using fibroblast cell line XPTNF2, which is an SV 40 fibroblast line deficient in DNA repair and carrying the TNF-a promoter chloramphenicol acetyl­ transferase (CAT) reporter gene. UVB irradiation of these cells increased the promoter activity, and as a result exhibited a CAT activity of 39.87 nmol/mg/h. Twenty µM and 50 µM EGT reduced CAT activities to 15.97 and 22.07 nmol/mg/h, respectively (Table II). MMP-1 SYNTHESIS INDUCED BY UVA IRRADIATION We examined the production of MMP-1 from UV A-irradiated fibroblasts. In order to

ANTIOXIDANT ACTIVITY OF EGT 23 (a) (b) (c) 326 328 330 332 334 Field (mT) Figure 4. Singlet oxygen quenching effect of EGT by ESR study. (a) Control (without EGT). (b) EGT 10 mg/ml. (c) EGT 20 mg/mJ. Table I Inhibition of EGT on Lipid Peroxidation Initiated by Singlet Oxygen Liposomes alone Li posomes + rose bengal Lip + rose bengal + 20 µM EGT nmol LPO/ml 23.81 91.84 26.53 As a source of singlet oxygen, the photo-irradiated rose bengal system was used. Liposomes prepared from phosphatidyl choline with 10 mM rose bengal were irradiated by using a Sylvania 150 W slide projector. Oxidation products in the liposomes were assayed with K-Assay1M LPO-CC from Kamiya Biomedical Company (Seattle, WA). Data are expressed as mean value from dependent examinations in duplicate. address the contribution of 1 O 2 to new MMP-1 production, the effect of NaN 3, a typical quencher against 1 0 2 , on UV A-induced MMP-1 production was examined. UV A ex­ posure to fibroblasts at 40 J/cm2 dramatically increased MMP-1 secretion by 70-fold in cultured medium (data not shown). Fibroblasts produced a small amount of MMP-1 (0.3 ± 2.3 units/cells) in sham­ irradiated cells. On the other hand, UV A-exposed fibroblasts irradiated with 20 J/cm2