Inhibitory effects of (-)-epigallocatechin-3-gallate on melanogenesis in ultraviolet A-induced B16 murine melanoma cell
Purpose: To investigate the anti-melanogenesis effect of green tea compound, (-)-epigallocatechin-3- gallate (EGCG), on B16 murine melanoma cell irradiated by ultraviolet A (UVA) in the search for natural skin-lightening alternative agents.
Methods: B16 murine melanoma cells by UVA (9.0 J/cm2) for 0 to 32 min and then incubated in Dulbecco's Modified Eagle's Medium (DMEM) with EGCG (0-200 μg/mL) for 2 days. Cell viability was determined by MTT method and cell protein was quantified using a PA102 Bradford protein assay kit. Activity of tyrosinase (TRY) was determined based on the oxidation rate of 3,4-dihydroxy phenylalanine (DOPA). The ultra-structure of the melanosomes was observed by transmission electron microscopy (TEM).
Results: TRY activity and melanin concentration were increased to 146.70 ± 10.28 % (p < 0.05) and 157.06 ± 6.37 % (p < 0.05), respectively, by 9.0 J/cm2 UVA irradiation for 8 min, compared to blank control without UV A and EGCG. EGCG inhibited the UV A induced increase in TRY activity and melanin level, and the optimum concentration of EGCG was 25 μg/mL. TRY activity and melanin concentration were decreased to 64.71 ± 4.41 (p < 0.05) and 86.24 ± 5.15 % (p < 0.05), respectively, compared to blank (control) which was neither treated by UVA nor by EGCG. TEM showed that UVA induced the formation of melanosomes while EGCG inhibited UVA-induced melanosome maturation.
Conclusion: EGCG inhibits UVA-induced melanogenesis via suppression of TRY activity and melanosome maturation and is thus a potential alternative to melanogenesis inhibitor.
Keywords: Green tea, Catechins, Melanin, Melanosome, Tyrosinase, Cell proliferation
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