ANTIOXIDANT ACTIVITY OF EGT 21 carried out for 3 5 cycles. PCR products were analyzed by 1 % agarose gel electrophoresis and visualized by ethidium bromide staining under ultraviolet light. RESULTS SCAVENGING ABILITY AGAINST •0 2 - The scavenging ability of EGT against •0 2 - was evaluated using the hypoxanthine and xanthine oxidase system as a source of •02 - . EGT showed scavenging against •02 - in a dose-dependent manner in the micromolar range (Figure 2). In addition, we examined the effects of EGT on lipid peroxidation (LPO) of liposomes initiated by •02 - generated by alloxan. The basal level of LPO in the control liposome was 17 .3 7 nmol/ml, and the addition of alloxan to the system was increased to 50.31 nmol/ml. EGT (20 µM) reduced LPO to 22.12 nmol/ml, an 85% reduction, and exhibited superior effects among other sulfur-containing antioxidants that were tested at the same concentration (Figure 3 ). QUENCHING ACTIVITY AGAINST 1 02 The quenching activity of EGT was measured by using the ESR spin-trapping method and lipid peroxidation (LPO) initiated by 1 0 2 . In general, hematoporphyrin produces 1 0 2 during UV A irradiation. As a source of 1 0 2 , the hematoprophyrin and UV A system was used. The ESR spectrum of the 1 0 2 is shown in Figure 4. The addition of EGT showed a decrease of 1 Orderived TEMP radicals in a dose-dependent manner. These results indicated that EGT effectively quenched 1 0 2 . The results for LPO initiated by 30 .-. 25 � en 20 C en 15 C 10 UJ 5 0 0 20 40 60 Cone. of EGT (µM) Figure 2. Scavenging effect of EGT against superoxide anion generated by hypoxanthine-xanthine oxidase system. The OD56onrn in the absence of EGT was set as 0% scavenging. The scavenging percent was calculated as the reduction in OD divided by the starting OD, x 100.

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