J Vet Sci.  2015 Jun;16(2):135-143. 10.4142/jvs.2015.16.2.135.

Anti-melanogenic effects of black, green, and white tea extracts on immortalized melanocytes

• Kim YC
• Choi SY
• Park EY

• KMID: 2164521
• DOI: http://doi.org/10.4142/jvs.2015.16.2.135

Abstract

Tea contains polyphenols and is one of the most popular beverages consumed worldwide. Because most tyrosinase inhibitors that regulate melanogenesis are phenol/catechol derivatives, this study investigated the inhibitory effects of Camellia sinensis water extracts (CSWEs), including black tea, green tea, and white tea extracts, on melanogenesis using immortalized melanocytes. CSWEs inhibited melanin accumulation and melanin synthesis along with tyrosinase activity in a concentration-dependent manner. These inhibitory effects were superior to those of arbutin, a well-known depigmenting agent. The anti-melanogenic activity of black (fermented) tea was higher than that of a predominant tea catecholamine, epigallocatechin gallate. CSWEs, especially black tea extract, decreased tyrosinase protein levels in a concentration-dependent manner. These results suggest that the anti-melanogenic effect of CSWEs is mediated by a decrease in both tyrosinase activity and protein expression, and may be augmented by fermentation. Thus, CSWEs could be useful skin-whitening agents in the cosmetic industry.

Keyword

Camellia sinensis; catecholamine; melanocytes; melanogenesis; tyrosinase

MeSH Terms

Animals
Catechin/analogs & derivatives/metabolism
Cell Line
Melanins/*metabolism
Melanocytes/enzymology/*metabolism
Mice
Monophenol Monooxygenase/*metabolism
Plant Extracts/*pharmacology
Plant Leaves/chemistry
Tea/*chemistry
Catechin
Melanins
Monophenol Monooxygenase
Plant Extracts
Tea

Figure

• 

  Fig. 1 Effect of different tea extracts on cell viability. Melan-A cells were treated with the indicated concentrations of the test reagents (0~50 µg/mL) for 48 h. Values represent the mean ± SD of three independent experiments. AT: arbutin, KA: kojic acid, EGCG: epigallocatechin gallate, BT: black tea, GT: green tea, WT: white tea.

• 

  Fig. 2 Anti-melanogenic effect of the tea extracts. (A) Morphological changes of melan-A cells treated with the extracts. The treatment with 12.5 µg/mL for 72 h reduced the amount of black precipitate and dendrite extension compared to the untreated control. BT at lower concentrations of 3.125 µg/mL (BT1) and 6.25 µg/mL (BT2) also significantly inhibited melanogenesis. (B) Melanin production in cells treated with the test reagents (12.5 µg/mL) for 72 h was measured with an ELISA. BT1 is 3.125 µg/mL, BT2 is 6.25 µg/mL, and BT3 is 12.5 µg/mL. Values represent the mean ± SD of three independent experiments. Statistically significant differences (p < 0.001) identified by an ANOVA and Duncan's multiple range test are indicated by different letters. *p < 0.05 compared to the untreated control. 200× magnification. C: control.

• 

  Fig. 3 Effect of the tea extracts on tyrosinase activity in melan-A cells. Tyrosinase activity in the cells treated with 12.5 µg/mL of the test reagents for 48 h was measured with an ELISA. BT1 is 3.125 µg/mL, BT2 is 6.25 µg/mL, and BT3 is 12.5 µg/mL. Values represent the mean ± SD of three independent experiments. Statistically significant differences (p < 0.001) identified by an ANOVA and Duncan's multiple range test are indicated by different letters.

• 

  Fig. 4 Effect of the tea extracts on tyrosinase protein expression in melan-A cells. Tyrosinase protein expression in cells treated with the test reagents (0~25 µg/mL) for 48 h was analyzed by Western blotting. (A) A representative gel. (B) Relative protein expression normalized relative to β-actin levels. Values represent the mean ± SD of three independent experiments. Treatment with BT, GT, or WT decreased tyrosinase protein expression in a concentration-dependent manner compared to the untreated control. Significant differences (p < 0.001) identified by an ANOVA and Duncan's multiple range test are indicated by different letters. **p < 0.01 compared to the untreated control.

• 

  Fig. 5 Effect of the tea extracts on mRNA expression of tyrosinase, TRP-1, and TRP-2 in melan-A cells. mRNA expression in cells treated with the test reagents (0~12.5 µg/mL) for 48 h was analyzed by RT-PCR. (A) A representative gel. (B) Semi-quantification of mRNA expression at 12.5 µg/mL by normalization to β-actin. Values represent the mean ± SD of three independent measurements. mRNA expression of tyrosinase, TRP-1, and TRP-2 was not affected by treatment with BT, GT, or WT.

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