• Proceedings of the SCSK Conference
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• 2003.09b
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• pp.241-242
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• 2003

The effects of N-acetylphytospingosine(NAPS), one of the phytospingosine derivatives, on melanogenesis of B 16F 1 0 mouse melanoma cell lines were investigated. We assessed the effect of NAPS on the depigmentation of B16F10 cells. The melanin content of cells was significantly reduced by NAPS. We examined the inhibitory effect of NAPS on tyrosinase activity using L-dopa as a substrate and the results showed that tyrosinase activity was inhibited in a does-dependent manner. The mRNA level of tyrosinase as well as that of tyrosinase related protein-l (TRP-l) and tyrosinase related protein-2 (TRP-2) genes were not affected by NAPS based on a reverse transcription-polymerase chain reaction (RT-PCR) assay. We also performed a Western blotting analysis using anti-tyrosinase antibody. It showed that there is no change in tyrosinase protein level after treatment of NAPS. These results suggest that the depigmenting mechanism of NAPS in B16F10 melanoma cells involves inhibition of melanosomal tyrosinase activity, rather than the mRNA expression or protein level of tyrosinase.

• Journal of Microbiology and Biotechnology
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• v.28 no.12
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• pp.2121-2132
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• 2018

Abnormal melanin synthesis results in several hyperpigmentary disorders such as freckles, melanoderma, age spots, and other related conditions. In this study, we investigated the anti-melanogenic effects of an extract from the microalgae Chlamydomonas reinhardtii (CE) and potential mechanisms responsible for its inhibitory effect in B16F10, normal human epidermal melanocyte cells, and human skin-equivalent models. The CE extract showed significant dose-dependent inhibitory effects on ${\alpha}$-melanocyte-stimulating, hormone-induced melanin synthesis in cells. Additionally, the CE extract exhibited suppressive effects on the mRNA and protein expression of microphthalmia-associated transcription factor, tyrosinase, tyrosinase-related protein-1, and tyrosinase-related protein-2. The CE extract also inhibited the phosphorylation of protein kinase A and extracellular signal-related kinase, which function as upstream regulators of melanogenesis. Using a three-dimensional, reconstructed pigmented epidermis model, the CE-mediated, anti-pigmentation effects were confirmed by Fontana-Masson staining and melanin content assays. Taken together, CE extract can be used as an anti-pigmentation agent.

• Mycobiology
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• v.40 no.2
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• pp.117-123
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• 2012

Nuruk contributes to the unique characteristics of Korean alcoholic beverages. In this study, the effects of nuruk extracts (NE) on anti-oxidant characters, melanogenesis, and anti-photoaging activity were investigated. NEs were obtained from the 70% ethanol extracts of six types of nuruk, which have been used in brewing of fermented alcohol beverages in Korea. First, various antioxidant characteristics were identified in terms of 2,2'-azino-bis(3-ethylbenzthiozoline-6-sulphonic acid) (ABTS) radical scavenging activity, superoxide dismutase (SOD) expression, and inhibition of xanthine oxidase activity. NE#4 exhibited potent ABTS radical scavenging activity ($IC_{50}$ = 19.51 ${\mu}g$/mL). Compared with NE#4, relatively lower levels of activity were observed for NE#3 and NE#6, with $IC_{50}$ values of 90.99 and 76.88 ${\mu}g$/mL, respectively. According to results of western blot analysis for determination of SOD expression in $H_2O_2$-treated HepG2 cells, NE#5 and NE#6 induced a dramatic increase in the expression ratio of SOD, compared to the group treated with $H_2O_2$ only. Activity of xanthine oxidase, which converts xanthine into uric acid, generating superoxide ions, was inhibited by NE#4 and NE#6 in a dose-dependent manner. NE#4 induced significant inhibition of mushroom tyrosinase activity. A reduction in cellular melanin contents of 80% was observed in B16F1 melanocytes treated with NE#5 and NE#6; these effects were similar to those of arbutin at 100 ${\mu}M$. In addition, gelatin zymography and reverse transcription-PCR analysis were performed for assessment of anti-photoaging activity of Nuruk. Treatment with NE#6 resulted in dramatically inhibited activities of matrix metalloproteinase (MMP)-2/-9, suppressed expression of MMP-1, and increased expression of type-1 procollagen. Results of gelatin zymography for NE#4 and NE#5 were similar, to a slightly lesser degree. These results suggest the potential of NE#4 and NE#6 as natural ingredients for use in functional foods and cosmetics.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.34 no.2
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• pp.101-107
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• 2008

In order to investigate the potential of a Angelica keiskei Koidzumi extract and its fractions as an active ingredient for whitening cosmetics, we prepared Angelica keiskei Koidzumi extract(70% ethanol), and the aqueous suspension was successively extracted with hexane, ethyl acetate(EA), and n-butanol fraction. We measured their inhibitory effects on mushroom tyrosinase and melanin synthesis in B16 melanoma cells and normal human kerationocytes in vitro. They did not show inhibitory activity against mushroom tyrosinase and the melanin synthesis except hexane and EA fractions. Hexane and EA fractions markedly inhibited cellular tyrosinase activity at a lower concentration(25 and 5 ${\mu}g/mL$, respectively) than arbutin(250 ${\mu}g/mL$). We also quantified the released amount of endothelin-1(ET-1), a mitogen of melanocyte, and interleukin-$1{\alpha}$(IL-$1{\alpha})$, a mediator of UVB-induced inflammation. Hexane and EA fractions did not affect IL-$1{\alpha}$ production, but they decreased UVB-induced ET-1 production in normal human keratinocytes in a dose dependant manner. As a result, hexane and EA fractions could inhibit the melanogenesis through the inhibition of UVB-induced ET-1 production in normal human keratinocytes. This result suggests that hexane and EA fractions of Angelica keiskei Koidzumi extract could be used as an active ingredient for cosmetics.

• Journal of Life Science
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• v.23 no.12
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• pp.1516-1524
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• 2013

To find a novel skin whitening agent, the effect of cordycepin-enriched Cordyceps militaris (CM${\alpha}$) extract fermented by fungi on anti-melanogenesis in B16F0 mouse melanoma cells was investigated. Fermented CM${\alpha}$ was prepared with fungi, including Monascus purpureus (Mp), Aspergillus oryzae (Ao), Aspergillus kawachii (Ak), and Rhizopus oryzae (Ro), respectively. When the content of the phenolics and the flavonoids and the activities of the antioxidant and the mushroom tyrosinase inhibition were measured in the CM fermented by Ak (AkF-CM), the highest content of the phenolics was 46 mg/g dry weight and the highest content of the flavonoids was 0.93 mg/g; the highest activity of the DPPH radical scavenging was 62.74% and the highest activity of the mushroom tyrosinase inhibition was 79.97% CM${\alpha}$CM${\alpha}$. From this result, AkF-CM${\alpha}$ exhibited the highest mushroom tyrosinase inhibitory activity and so it was used in subsequent anti-melanogenesis. B16F0 melanoma cells were treated with 1-10 mg/ml concentrations of AkF-CM${\alpha}$ and 200 ${\mu}M$ arbutin as the positive control. The melanin content and cell viability of the melanoma cells by arbutin treatment decreased to 43% and 92% of the control, respectively. AkF-CM${\alpha}$ treatment at 1, 3, and 5 mg/ml concentrations decreased the extracellular melanin release induced by IBMX treatment by 35%, 45%, and 53%, respectively. AkF-CM${\alpha}$ showed inhibitory activity against both intracellular tyrosinase in melanoma cells and mushroom tyrosinase. AkF-CM${\alpha}$ reduced the protein level of tyrosinase in the IBMX-stimulated cells. These results indicate that AkF-CM${\alpha}$ suppressed the activity and protein content of cellular tyrosinase and decreased the total melanin content in cultured B16F0 melanoma cells.

• Nutrition Research and Practice
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• v.11 no.3
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• pp.173-179
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• 2017

BACKGROUND/OBJECTIVES: Gastrodia elata Blume (GEB), a traditional herbal medicine, has been used to treat a wide range of neurological disorders (e.g., paralysis and stroke) and skin problems (e.g., atopic dermatitis and eczema) in oriental medicine. This study was designed to investigate whether GEB extract inhibits melanogenesis activity in murine B16F10 melanoma. MATERIALS/METHOD: Murine B16F10 cells were treated with 0-5 mg/mL of GEB extract or $400{\mu}g/mL$ arbutin (a positive control) for 72 h after treatment with/without 200 nM alpha-melanocyte stimulating hormone (${\alpha}$-MSH) for 24 h. Melanin concentration, tyrosinase activity, mRNA levels, and protein expression of microphthalmia-associated transcription factor (MITF), tyrosinase, tyrosinase-related protein (Trp)1, and Trp2 were analyzed in ${\alpha}$-MSH-untreated and ${\alpha}$-MSH-treated B16F10 cells. RESULTS: Treatment with 200 nM ${\alpha}$-MSH induced almost 2-fold melanin synthesis and tyrosinase activity along with increased mRNA levels and protein expression of MITF, tyrosinase, Trp1 and Trp2. Irrespective of ${\alpha}$-MSH stimulation, GEB extract at doses of 0.5-5 mg/mL inhibited all these markers for skin whitening in a dose-dependent manner. While lower doses (0.5-1 mg/mL) of GEB extract generally had a tendency to decrease melanogenesis, tyrosinase activity, and mRNA levels and protein expression of MITF, tyrosinase, Trp1, and Trp2, higher doses (2-5 mg/mL) significantly inhibited all these markers in ${\alpha}$-MSH-treated B16F10 cells in a dose-dependent manner. These inhibitory effects of the GEB extract at higher concentrations were similar to those of $400{\mu}g/mL$ arbutin, a well-known depigmenting agent. CONCLUSIONS: These results suggest that GEB displays dose-dependent inhibition of melanin synthesis through the suppression of tyrosinase activity as well as molecular levels of MITF, tyrosinase, Trp1, and Trp2 in murine B16F10 melanoma. Therefore, GEB may be an effective and natural skin-whitening agent for application in the cosmetic industry.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.40 no.4
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• pp.403-412
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• 2014

To develop an effective skin whitening agent for cosmetics, we isolated cucurbitacin B from Cucumis sativus L. which has been used as traditional skin lighting regimen by the bioactivity-guided fractionation, and investigated the inhibitory effects of cucurbitacin B on melanogenesis. At a non-cytotoxic concentration, cucurbitacin B reduced melanin contents of B16F1 melanoma cells in a dose-dependent manner. Cucurbitacin B did not directly inhibit mushroom tyrosinase activity, but it inhibited intracellular tyrosinase activity in a dose-dependent manner. Its inhibitory mechanism on melanin biosynthesis was further assessed, and we found that cucurbitacin B significantly decreased the protein level of tyrosinase, a major melanogenic enzymes and MITF, a master transcriptional factor of melanogenesis. In addition, cucurbitacin B increased the expression of WW domain-containing oxidoreductase (WWOX) which is known to function as tumor repressor and inhibits $Wnt/{\beta}$-catenin pathway. Collectively, these results suggest that cucuritacin B from C. sativus could be used as an active ingredient for skin whitening.

• Korean Journal of Pharmacognosy
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• v.33 no.4 s.131
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• pp.399-403
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• 2002

Three melanogenesis inhibitors were isolated from the roots and rhizomes of Veratrum nigrum L. and were identified as (3S,20S,25S)-22,26-iminocholesta-5,22(N)-dien-3-ol (verazine), (3S,2OR,25S)-22,26-iminocholesta-5,22(N)-dien-3-ol (epi-verazine) and (3R,23R)-14,15,16,17- tetradehydroveratraman-3,23-diol (veratramine) on the basis of their spectroscopic data. It was turned out that these compounds did not directly inhibit tyrosinase activity, the key enzyme responsible for the formation of melanin pigment while these compounds showed strong inhibition on the melanogenesis in B16 F1 mouse melanoma $(IC_{50}<1\;{\mu}g/ml)$. Due to the strong inhibitory activity and safety compared to current whitening agents such as arbutin, kojic acid and AHA, the compound can be a good candidate for new skin whitening agents.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.33 no.2
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• pp.87-92
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• 2007

To develop the skin whitening agent, we investigated the effects of Acanthopeltis japonica, a rhodophyta on the coast of Jeju island, on melanogenesis. Dried A. japonica was refluxed with 70 % aqueous ethanol and the extract was evaporated to dryness. To validate the activity as a depigmenting agent, various in vitro tests, polyphenol contents, and free radical scavenging activity were performed. In addition, cellular tyrosinase activity and protein expression of p-ERT, tyrosinase, TRP-1, and TRP-2 were measured in B16/F10 murine melanoma cells. A. japonica had low polyphenol contents and low free radicals scavenging activities against 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical. A. japonica suppressed cellular tyrosinase activity up to 86.9 % at $100{\mu}g/mL$ with inhibition or tyrosinase and TRP-1 expression in ${\alpha}$-melanocyte stimulating hormone (${\alpha}$-MSH)-treated B16/F10 melanoma cells. Our results suggest that inhibitory effects of A. japonica on melanogenesis are due to inhibiting the pathways involving ${\alpha}$-MSH-induced ERK activation. Therefore, A. japonica nay be useful as a skin whitening agent associated with the suppressive effect of melanotrophin-induced signaling pathway to inhibit melanin synthesis.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.40 no.4
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• pp.341-347
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• 2014

This study was aimed to investigate skin whitening effects of Nomura's jellyfish (Nemopilema nomurai) hydrolyzed extracts (NHE). The extracts were prepared through the hydrolysis of N. nomurai using commercial proteolytic enzymes such as Protamex, Alcalase, Flavourzyme and Neutrase with optimum pHs (pH 5-8) at $55^{\circ}C$. Their whitening activities were examined from the inhibition of melanin synthesis using B16-F1 cell lines. Among the examined samples, Neutrase-treated extract (N-NHE) showed the most significant inhibition effect on melanin synthesis by 89.9% at a concentration of $100{\mu}g/mL$. This sample decreased the expression of tyrosinase and TRP-1 (tyrosinase related protein-1) proteins as well. These results suggested the potential application of NHE as whitening ingredients in cosmetic preparation.