• Journal of the Society of Cosmetic Scientists of Korea
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• v.23 no.2
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• pp.23-32
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• 1997

Melanin pigmentation in human skin is a major defense mechanism against ultraviolet light of the sun. Tyrosinase(EC 1.14.18.1) plays a key role in the biosynthesis of ultraviolet of melanin. This is why much researches have been focused on its regulation in controlling the epidermal melanization. We have found that the water-extract of Banha(Pinelliae ternate B.), an oriental medicinal plant, has no tyrosinase inhibitory activity, but does inhibit the melanin biolsynthesis in B16 mouse melanin cells. We also found that Banha extract lowers the tyrosinase activity in cultured cells. To elucidate the action mechanism of Banha extract we have investigated its effect on the tyrosinase mRNA level using reverse transcription-polymerase chain reaction technique. It was revealed that Banha extract reduced the tyrosinase mRNA level in dose dependent manner; when B16 mouse melanoma cells were cultured with 2mg/ml and 5mg/ml of Banha extract, there were 20% and 44% decrease in tyrosinase mRNA level, respectively. These data suggest that the Banha extract exerts its melanogenic inhibitory effect through the transcriptional regulation of tyrosinase mRNA.

• Korean Journal of Food Science and Technology
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• v.48 no.1
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• pp.72-76
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• 2016

We investigated the application of functional materials by examining a variety of physiological activities of Geranium krameri extract obtained from the National Institute of Horticultural and Herbal Science. Geranium krameri extract had a low cytotoxicity against murine melanoma B16F10 cells. At concentrations with little or no cytotoxicity, Geranium krameri extract showed high DPPH radical scavenging activity ($IC_{50}$, $8.72{\mu}g/mL$) and anti-microbial activities against Bacillus subtilis, Escherichia coli, and Candida albicans. Additionally, Geranium krameri extract inhibited tyrosinase activity ($IC_{50}$, $456.86{\mu}g/mL$) and decreased melanin content ($IC_{50}$, $50.35{\mu}g/mL$). The treatment of B16F10 cells with Geranium krameri extract suppressed the protein expression of tyrosinase in a dose-dependent manner. These findings suggest that Geranium krameri extract inhibits melanin synthesis in murine melanoma B16F10 cells by suppressing intracellular tyrosinase expression, as well as directly inhibits tyrosinase activity simultaneously.

• Korean Journal of Food Science and Technology
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• v.53 no.2
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• pp.160-164
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• 2021

This study investigated various physiological activities to examine the applicability of the functional materials of Allium tuberosum root extract. The A. tuberosum root extract showed a low cytotoxicity against murine melanoma B16F10 cells. It also showed high DPPH radical scavenging activity (ID50, 6.2 ㎍/mL), inhibited tyrosinase activity (ID50, 115.4 ㎍/mL), and decreased melanin content (ID50, 31.5 ㎍/mL). Treatment of B16F10 cells with A. tuberosum root extract suppressed the protein expression of tyrosinase in a dose-dependent manner. These findings suggest that A. tuberosum root extract inhibits melanin synthesis by suppressing intracellular tyrosinase expression. Additionally, A. tuberosum root extract inhibited elastase with an ID50 value of 145.1 ㎍/mL and contained isoquercitrin. These results indicate that A. tuberosum root extract is an appropriate natural material.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.37 no.4
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• pp.319-326
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• 2011

In order to investigate the potential of Nigella sativa (N. sativa) oil as an active ingredient for whitening cosmetics, we prepared N. sativa oil. We measured its inhibitory effects on mushroom tyrosinase activity, cellular tyrosinase activity, and melanin synthesis inhibitory activity in B16 melanoma cells. N. sativa oil and its components showed inhibitory activity against mushroom tyrosinase and melanin synthesis. In a melanin synthesis inhibition assay using mouse B16-F10 melanoma cell, it reduced melanin production up to 86 % at a concentration of 10 mg/mL without cytotoxicity. In the study on the melanogenic protein expressions by using RT-PCR and Western blot, N. sativa oil and its components inhibited expression of tyrosinase protein, which is a well-known key protein on melanogenesis, and tyrosinase expression was gradually decreased in a dose-dependent manner. Therefore, this result suggests that N. sativa oil could be used as an active ingredient for whitening cosmetics.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.38 no.3
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• pp.247-254
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• 2012

Ramalin (${\gamma}$-glutamyl-N'-(2-hydroxyphenyl)hydrazide) isolated from the Antarctic lichen Ramalina terebrata has been shown to have strong antioxidant activities in the previous study. To investigate additional activities of ramalin, we studied the effects of ramalin on melanogenesis in melan-a cells, a non-tumorigenic melanocyte cell line. At a non-cytotoxic concentration, ramalin dramatically decreased melanin synthesis in melan-a cells in a dose-dependent manner, which was more potent than arbutin, a well-known tyrosinase inhibitor. Ramalin inhibited cell-free tyrosinase activity directly and intracellular tyrosinase activity as well. Its inhibitory mechanisms on melanin production were further assessed, and we found that ramalin significantly decreased the protein levels of melanogenic enzymes such as tyrosinase, tyrosinase-related protein 1 (TRP-1), and tyrosinase-related protein 2 (TRP-2). However, the mRNA levels of these enzymes were not altered. In a clinical study, application of 0.2 % ramalin on human skin significantly improved the degree of skin brightness after 3 weeks. In conclusion, ramalin has strong anti-melanogenic activity that is exerted both by the direct inhibition of tyrosinase activity and by down-regulation of melanogenic proteins. Furthermore, ramalin showed skin brightening effect in a clinical study. Collectively, these results suggest that ramalin may be a useful inhibitor for melanogenesis in skin.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.38 no.1
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• pp.67-73
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• 2012

In the present study, we investigated the effects of dipeptides on melanogenesis in B16 melanoma cells. It was found that WV (Trp+Val), WM (Trp+Met), and CQ (Cys+Gln) decreased melanin production dosedependently. However, dipeptides did not directly inhibit tyrosinase activity, the rate-limiting melanogenic enzyme. Therefore, we further investigated the expression of tyrosinase. Our results showed that ${\alpha}$-MSH-induced tyrosinase expression was down-regulated by WV, WM, and CQ. Thus, we propose that WV, WM, and CQ show hypopigmentary activity through tyrosinase down-regulation.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.31 no.1 s.49
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• pp.17-23
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• 2005

We had previously reported that Selina (selina-4(14), 7(11)-dien-8-one) was isolated from methanol extract of Afractylodes rhizome and has strong whitening activity in B16 melanoma cells. In this report, we demonstrated its action mechanism in melan-a cells, non-tumorigenic melanocytes. We also investigated the clinical efficacy of cosmetic preparation containing Selina. Selina reduced the melanin synthesis of Melan-a cells by $50\%$ at a concentration of $10 {\mu}g/mL$ without any apparent cytotoxicity. We also found that the treatment of cells with Selina decreased tyrosinase activity by $60\%$ at a concentration of $10 {\mu}g/mL$ but Selina was not a direct inhibitor of tyrosinase activities. To elucidate the action mechanism of Selina, we investigated the changes in mRNA and protein level of tyrosinase, TRP-1 and TRP-2 using RT-PCR and western blotting, respectively. As a result, the mRNA and protein level of tyrosinase were markedly reduced at $10 {\mu}g/mL$ of Selina without any effect on TRP-1 and TRP-2. These results suggest that Selina exerts its whitening effect mainly through regulating expression of tyrosinase. A 7 week-clinical trial using formulation containing $0.2\%$ selina-4(14), 7(11)-dien-8-one with 20 volunteers resulted in statistically significant whitening effect (p < 0.05), without any adverse effect. Based on these results, Selina (selina-4(14), 7(11)-dien-8-one) can be s useful and safe ingredient for the cleanness and brightness of skin.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.31 no.4 s.54
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• pp.305-310
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• 2005

To obtain effective and safe depigmenting agents, we investigated the effects of Scirpi rhizoma, a medicine among Chinese herbs, on melanogenesis. Dried S. rhizoma was refluxed with 70% aqueous ethanol and the extract was evaporated to dryness. To determine the effects as a whitening agent, various in vitro tests were performed such as free radical scavenging activity, melanin formation assay, tyrosinase activity and expression of tyrosinase, TRP-1 and TRP-2(western blot and RT-PCR) in B16 melanoma cells. S. rhizoma showed scavenging activities of free radicals and reactive oxygen species (ROS) with the $IC_{50}\;of\;638{\mu}g/mL$ against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and $21.7{\mu}g/mL$ against superoxide radicals in the xanthine/xanthine oxidase system, respectively. S. rhizoma significantly inhibited melanin production in B16 melanoma cells. S. rhizoma treatment(48 h) suppressed the biosynthesis of melanin up to 27% at 100{\mu}g/mL$ and reduced tyrosinase activity up to 31% at $100{\mu}g/mL$ in B16 melanoma cells. S. rhizoma was also able to significantly inhibit tyrosinase and TRP-1 expression in protein and mRNA level. These results suggest that S. rhizoma inhibited melanin biosynthesis by regulating tyrosinase activity and expression in B16 melanoma cells. Therefore, S. rhizoma may be useful as a new antioxidant and whitening agent to inhibit melanogenesis.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.48 no.1
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• pp.11-24
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• 2022

It has been reported that the ethanolic extract of the root of Piper methysticum (P. methysticum) inhibits melanogenesis in melanocyte stimulating hormone (MSH)-activated B16 melanoma cells. Flavokawain B (FKB) and Flavokawain C (FKC) isolated from this extract have been found to inhibit melanin production based on anti-melanogenesis activity. This study was designed to find out the inhibition and its process of FKB and FKC on melanin synthesis in melan-a melanocytes. FKB and FKC inhibited melanogenesis at 10 μM, 5 μM respectively in melan-a melanocytes. However, they did not inhibit extracellular tyrosinase activity from melan-a melanocytes. FKB reduced the protein level of tyrosinase (Tyr), tyrosinase-related protein 1 (TRP-1), tyrosinase-related protein 2 (TRP-2), microphthalmia-associated transcription factor (MITF) and the mRNA level of Tyr and TRP-1. FKC reduced the protein level of TRP-2 and MITF and the mRNA level of TRP-1 and Tyr. The reduced expression of Tyr and TRP-1 might be resulted from the decreased MITF which regulates major melanogenic proteins. However, since the mRNA expression of MITF did not change by FKB and FKC treatment, the effects of FKB and FKC on extracellular signal regulating kinase (ERK)/AKT phosphorylation, known to regulate the degradation of MITF, were confirmed. FKB and FKC significantly increased the phosphorylation of ERK1/2, not in AKT. These results suggest that FKB and FKC may be helpful as a potential depigmenting agent for various hyper-pigmentary disorders.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.34 no.3
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• pp.183-188
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• 2008

Initial screening assay for depigmenting agents includes in vitro mushroom tyrosinase assay and antioxidant assay. Based on this screening result, melanin synthesis in melanocyte, in screened samples, is further measured. Measuring cellular melanin needs time, human resource, and skills. Therefore initial screening method should be reliable. We examined, 34 Chinese herbs, correlated the screening assay methods with cellular melanin. No reliable relationship was observed between factors, indicating the limitation in the use of these assays, probably due to the complexicity of melanogenesis.