• YAKHAK HOEJI
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• v.52 no.3
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• pp.165-171
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• 2008

To develop effective skin whitening agents, we tested natural herbal extracts for their melanogenic inhibitory activities. Sedum samentosum was selected for its inhibitory effect on melanogenesis in B16 melanoma cells. Ethanolic extract of S. samentosum (SSE) was evaluated for antioxidative effect and tyrosinase inhibitory activity of melanogenesis. We investigated the changes in protein level and mRNA level of tyrosinase, tyrosinase related protein (TRP)-1 and TRP-2 by using western blotting and RT-PCR, respectively. SSE showed scavenging activities of free radicals and reactive oxygen species (ROS) with the $IC_{50}$ of 342.7 $\mug/ml$ against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and 64.69 $\mug/ml$ against superoxide radicals in the xanthine/xanthine oxidase system, respectively. SSE treatment suppressed the biosynthesis of melanin up to 46% and reduced tyrosinase activity up to 51% at 100 $\mug/ml$ in B16 melanoma cells. The tyrosinase activity and tyrosinase expression in B16 melanoma cells were reduced in a dose-dependent manner by SSE. Also, SSE was able to significantly inhibit tyrosinase and TRP-1 expression in mRNA level. These results suggest that SSE inhibited melanin production which may be dependent on tyrosinase activity and expression in B16 melanoma cells, and an effective whitening agent for the skin.

• Animal cells and systems
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• v.10 no.4
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• pp.233-236
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• 2006

For the purpose of the development of skin-whitening or therapeutic agents against hyperpigmentation, aqueous extract from Nardostachys chinensis (AENC) was evaluated for melanogenesis inhibitory activity in B16F10 melanoma cell. The treatment with AENC at the 0.2, 0.5 and 1.0 mg/ml level significantly inhibits the biosynthesis of melanin compared with untreated control. The tyrosinase activity also significantly decreased in AENC-treated cells at the 0.2 and 0.5 mg/ml level and inhibitory effects were more efficient than commercial arbutin at 0.1 mg/ml. The Western analyses confirmed the significantly decreased expression of tyrosinase and tyrosinase-related protein-2 by AENC treatment. These results indicate that AENC may contribute to the inhibition of melanin biosynthesis through regulating the expression as well as activity of tyrosinase and AENC may be useful as a new candidate in the design of new skinwhitening or therapeutic agents.

• Journal of Physiology & Pathology in Korean Medicine
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• v.23 no.2
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• pp.337-342
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• 2009

Acquired pigmentary skin diseases such as abnormal melanogenesis, vitiligo, chloasma and inflammatory pigmentation are related to regulate the melanin production, In this study, an ethanol extract of Hovenia dulcis Thunb.(EHD) makedly inhibited melanin biosynthesis and suppressed, the protein expression of tyrosinase, tyrosinase-related protein 1(TRP-1), and tyrosinase-related protein 2(TRP-2) in B16F10 cells. On the other hand, EHD did not inhibit mushroom tyrosinase activity. These results indicate that EHD may contribute to the inhibition of melanin biosynthesis through regulating tyrosinase activity and expression, and serve as a new candidate in the design of new skin-whitening or therapeutic agents.

• Korean Journal of Acupuncture
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• v.37 no.1
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• pp.24-30
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• 2020

Objectives : The purpose of this study was to investigate melanogenesis inhibition of ethanol extract of Polyporus (EP) by using B16F10 melanoma cells. Methods : We measured antioxidant effect of EP by using 1,1-Diphenyl-1-picrylhydrazyl (DPPH) assay and we confirmed melanin contents and tyrosinase activity of EP in cells. Additionally, the expression of tyrosinase-related protein-1 (TRP-1) and TRP-2 was observed by Western blot. Results : EP showed significantly high radical scavenging activity and inhibition of melanogenesis in dose-dependent manner by decreasing cellular tyrosinase activity and melanin content with or without α-melanin stimulating hormone. TRP-1 and TRP-2 expressions were also suppressed by EP in B16F10 cells. Conclusions : These results suggest that EP inhibits the melanogenesis and it could be a new organic ingredient for hyper-pigmentation.

• Journal of Physiology & Pathology in Korean Medicine
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• v.21 no.5
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• pp.1243-1249
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• 2007

The aim of this study was to investigate the effect of ethanol extract of Fagopyrum esculentum on the melanogenesis. To determine whether ethanol extract of Fagopyrum esculentum suppress melanin synthesis in cellular level, B16F10 melanoma cells were cultured in the presence of different concentrations of Fagopyrum esculentum ethanol extract. In the present study, we examined the effects of Fagopyrum esculentum ethanol extract on cell proliferation, melanin contents, tyrosinase activity, expression of melanogenic enzyme proteins including tyrosinase, tyrosinase-related protein 1 (TRP-1) and tyrosinase-related protein 2 (TRP-2). Cell proliferation was slightly increased by treatment with ethanol extract of Fagopyrum esculentum $(25-200 {\mu}g/m{\ell}).$ The ethanol extract of Fagopyrum esculentum effectively suppressed melanin contents at a dose of $100 {\mu}g/m{\ell}).$ It was observed that the color of cell pellets was totally whitened compared with the control. The ethanol extract of Fagopyrum esculentum inhibited tyrosinase activity, regulate melanin biosynthesis as the key enzyme in melanogenesis. Using western blot analysis, the ethanol extract of Fagopyrum esculentum dose-dependently decreased tyrosinase and TRP-1 protein levels, and tyrosinase and TRP-1 were detected in similar manner. ${\alpha}-MSH$ leads to a stimulation of melanin synthesis through increase of tyrosinase activity, melanin contents and cytoplasmic dendricity. In this study, ethanol extract of Fagopyrum esculentum down-regulated the ${\alpha}-MSH$-induced tyrosinase activity, melanin contents and cytoplasmic dendricity. Regarding protein levels of the melanogenic enzymes, the amounts of tyrosinase and TRP-1 was increased after incubation with a-MSH. The treatment of ethanol extract of Fagopyrum esculentum decreased the ${\alpha}-MSH$-induced expression levels of tyrosinase and TRP-1. These results suggest that the ethanol extract of Fagopyrum esculentum exerts its depigmenting effects through the suppression of tyrosinase, TRP-1 and cytoplasmic dendricity. And it may be a potent depigmetation agent in hyperpigmentation condition.

• Microbiology and Biotechnology Letters
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• v.44 no.1
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• pp.1-8
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• 2016

The purpose of this study was to assess the whitening effects of an extract from Smilax china L., which is a vine shrub belonging to the lily family. With regard to the whitening effects, 70% ethanol and water extracts from Smilax china L. showed more than 77.6% and 40.2% tyrosinase inhibition at a concentration of $1,000{\mu}l$. Furthermore, the 70% ethanol extract showed cytotoxicity of 89% at a concentration of $100{\mu}g/ml$ in melanoma cells. Western blot showed that the inhibitory effect of the 70% ethanol extract on MITF, TRP-1, TRP-2, and tyrosinase protein expression decreased by 89.9%, 46.2%, 57.6%, and 55.8%, respectively, at a concentration of $50{\mu}g/ml$. Moreover, reverse transcription-PCR showed that the inhibitory effect of the 70% ethanol extract on MITF, TRP-1, TRP-2, and tyrosinase mRNA expression decreased by 78.5%, 58.0%, 78.8%, and 70.8%, respectively, at the same concentration of $50{\mu}g/ml$ concentration. Further, realtime PCR showed that the 70% ethanol extract-induced decrease in MITF, TRP-1, TRP-2, and tyrosinase quantitative mRNA expression rate was concentration-dependent. The findings suggest that the extract from Smilax china L. has great potential as a cosmetic ingredient with whitening effects.

• Journal of Life Science
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• v.24 no.5
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• pp.485-490
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• 2014

Poria cocas has been reported to be effective in skin whitening. However, the direct effect of P. cocas ethanol extracts (PCEE) on melanin synthesis has not been scientifically studied. To elucidate the direct effect of PCEE on melanogenesis, a 3,4-dihydroxyindole-2-carboxylic acid (DOPA) synthesis assay, tyrosinase activity assay, and Western blotting for melanogenic proteins, including tyrosinase, tyrosinase-related protein (TRP)-1, and TRP-2 were performed in mouse B16F1 cells. The results revealed that PGEE inhibited melanin production in a dose-dependent manner by blocking the synthesis of DOPA. Although the activation of tyrosinase was not affected, the expression levels of TRP-1 and TRP-2 were controlled. These results suggest that PCEE has a whitening effect, indicating that it may be a useful agent in the development of whitening cosmetics.

• The Korea Journal of Herbology
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• v.30 no.4
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• pp.95-100
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• 2015

Objectives : The purpose of this study was to research the whitening effects and developing by cosmetics of the extract fromDioscoreae Rhizoma, which is one of the most popular health-promoting herb in herbal medications.Methods : We performed tyrosinase inhibition assay, reverse transcription-polymerase chain reaction (RT-PCR) and western blot for whitening effects. Also we measured MTT assay for cell viability.Results : The results were obtained as follows : For whitening effect, tyrosinase inhibition rate of extract fromDioscoreae Rhizomashowed more than 42.28% at 1,000 ㎍/㎖ concentration. Cell toxicity effect on melanoma cells (B16F10) of extract fromDioscoreae Rhizomashowed 81.97% with toxicity at 50 ㎍/㎖ concentration. So we were measured at a concentrations of 5, 10 and 50 ㎍/㎖ in all experiments involving cell. In addition, whitening related mRNAs including microphthalmia associated transcription factor (MITF), tyrosinase related protein-1 (TRP-1), tyrosinase related protein-2 (TRP-2), tyrosinase were reduced byDioscoreae Rhizoma. We also foundDioscoreae Rhizomatransiently decreased protein kinase A (PKA) which is known to be upstream to the down regulation of MITF and tyrosinase. But phosphorylation of extracellular signal related kinase (pERK) were increased byDioscoreae Rhizoma. These results imply thatDioscoreae Rhizomadecrease melanogenesis via ERK activation and subsequent down regulation of MITF and tyrosinase.Conclusions : Therefore, all these findings suggested the potent usage ofDioscoreae Rhizomaas materials of functional cosmetics by confirming whitening activity related with melanin content.

• Korean Journal of Medicinal Crop Science
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• v.19 no.3
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• pp.177-184
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• 2011

This study was performed to assess the antioxidant activities and whitening effects of Agrimonia pilosa Ledeb on melanin synthesis. The whitening effects of Agrimonia pilosa Ledeb water extracts were examined by in vitro mushroom tyrosinase assay and B16BL6 melanoma cells. We assessed inhibitory effect of Agrimonia pilosa Ledeb water extract on expression of melanogenic enzyme proteins including tyrosinase, tyrosinase-related protein 1 (TRP-1) and tyrosinase-related protein 2 (TRP-2) in B16BL6 cells. Inhibitory effect of Agrimonia pilosa Ledeb onto free radical generation was determined by measuring DPPH and hydroxyl radical scavenging activitie. Our results indicated that Agrimonia pilosa Ledeb water extract effectively inhibited free radical generation. In DPPH and hydroxy radical scavenging activity, Agrimonia pilosa Ledeb water extract had a potent anti-oxidant activity in a dose-dependent manner. They significantly inhibited tyrosinase activity in vitro and in B16BL6 melanoma cells. Also, Agrimonia pilosa Ledeb suppressed the expression of tyrosinase in B16BL6 melanoma cells. These results show that Agrimonia pilosa Ledeb inhibited melanin production on the melanogenesis. The underlying mechanism of Agrimonia pilosa Ledeb on whitening activity may be due to the inhibition of tyrosinase activity. We suggest that Agrimonia pilosa Ledeb may be useful as new natural active ingredients for antioxidant and whitening cosmetics.

• Journal of Applied Biological Chemistry
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• v.54 no.4
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• pp.270-278
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• 2011

The objective of the present study was to evaluate the skin whitening effect of the extracts, Lycii fructus (LF), Dry-L. foilum (DLF) and Fresh-L. folium (FLF). Tyrosinase inhibition activities was 44% in DLF ethanol extracts at a $500{\mu}g/mL$. When the tyrosinase activities in B16F10 murine melanoma cell were tested, the activities in DLF ethanol extracts was 14% at a $50{\mu}g/mL$ concentration. The protein expression of microphthalmia-associated transcription factor, tyrosinase related protein 1 (TRP-1), TRP-2, and tyrosinase, which are all melanin related factors, showed that LF, DLF and FLF extracts inhibited the protein bio-synthesis in B16F10 melanoma cell. Especially the DLF extract showed greater decrease of protein expressions. Results indicate that the DLF extract tested in the present study had skin whitening activity and can be used as a function a ingredients for food and cosmetic compositions.