• Journal of Life Science
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• v.23 no.11
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• pp.1336-1341
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• 2013

The purpose of this study was to research the whitening effects of the extract from Angelica gigas Nakai, which is one of the most widely used herbal medicines in Asia. For whitening effects, the tyrosinase inhibition effect of the A. gigas Nakai extract was shown to be greater than 70% at 1,000 ${\mu}g/ml$ concentration. The result of measuring the cell toxicity effect of the extract from A. gigas Nakai on melanoma cells showed 99% toxicity at 500 ${\mu}g/ml$ concentration. The microphthalmia-associated transcription factor (MITF), tyrosinase related protein-1 (TRP-1), tyrosinase related protein-2 (TRP-2), and tyrosinase mRNA expression inhibitory effect by reverse transcription-PCR of the extract from A. gigas Nakai were decreased by 85.7%, 123.9%, 68.8%, and 208%, respectively, at 50 ${\mu}g/ml$ concentration. All these findings could verify that extract from A. gigas Nakai could have an effect on whitening. Moreover, extract from A. gigas Nakai has great potential as a cosmetic ingredient.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.43 no.3
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• pp.231-237
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• 2017

This study was performed to cytotoxicity, tyrosinase inhibition activity, intracellular melanin contents to verify the whitening effect of fraction from Glycine soja Siebold et Zucc. (G. soja). Using western blotting, tyrosinase expression in B16F10 melanoma cells and expression levels of tyrosinase related protein-1 (TRP-1) and protein-2 (TRP-2) were examined. As a result, all of the fractions showed a high cell viability over 82% at the concentrations of 0.125, 0.25, 0.5, 2.0 mg/mL. When the whitening effects of fractions from G. soja were tested using B16F10 melanoma cells treated with the ${\alpha}$-melanocyte stimulating hormone (${\alpha}-MSH$), the EtOAc fractions inhibited tyrosinase and melanogenesis effectively. The result of protein expression measurement using western blot showed that TRP-1, TRP-2 and tyrosinase protein expression in B16F10 melanoma cells treated with extracts decreased. Therefore, it is concluded that the fractions from G. soja have whitening effect by inhibiting protein related melanogenesis.

• 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.

• YAKHAK HOEJI
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• v.47 no.1
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• pp.31-36
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• 2003

To investigate the effect of protein kinase on melanin production via cAMP-dependent pathway, we measured the melanin amount and tyrosinase activity in B16 melanoma cells stimulated by alpha-melanocyte stimulating hormone (MSH), forskolin and 8-Br-cAMP. MSH, forskolin and 8-Br-cAMP significantly increased both melanin production and tyrosinase activity in B16 cells. Melanin production and tyrosinase activity by MSH are significantly inhibited by cyclic AMP-dependent protein kinase inhibitor (KT5720) and protein kinase C down-regulation treated with PMA. Bisindolmaleimide (1$\mu$M), protein kinase C inhibitor, significantly inhibited melanin production and tyrosinase activity stimulated by MSH, forskolin and 8-Br-cAMP with the following order of potency: MSH＞forskolin＞8-Br-cAMP. Tyrosine kinase inhibitor, genistein and DHC, significantly inhibited both, but the inhibitory effect was more potent in 8-Br-cAMP-stimulated B16 cells than MSH-stimulated cells. NFkB inhibitor (parthenolide) significantly inhibited melanin production and tyrosinase activity. Neither melanin production nor tyrosinase activity induced by MSH, forskolin and 8-Br-cAMP were affected by KN-62 (calmodulin-dependent protein kinase II inhibitor), PD098059 (mitogen-activated protein kinase inhibitor, MAPKK) and worthmannin (phosphatidylinositol 3-kinase inhibitor). These results suggest that both protein kinase C and tyrosine kinase are involved in melanin production by cyclic AMP-dependent pathway and NFkB pathway may play an important role in cyclic AMP-dependent melanin production in B16 melanoma cells.

• Journal of Physiology & Pathology in Korean Medicine
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• v.31 no.4
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• pp.226-232
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• 2017

In this study, ethanol extract of Epimedium koreanum Nakai(EEKN) enhanced melanogenesis by inducing expression of tyrosinase and tyrosinase-related protein-1 (TRP-1). But EEKN did not increase the protein expression of tyrosinase-related protein 2 (TRP-2). Moreover, EEKN enhanced tyrosinase activity and melanin contents of B16F10 cells. EEKN raised the expression of CREB phosphorylation and microphthalmia-associated transcription factor (MITF) as a key transcription factor for tyrosinase expression regulating melanogenesis. And PKC inhibitor H89 supressed that EEKN induced tyrosinase activity, melanin contents, and expression of tyrosinase, TRP-1. These results suggest that melanogenesis-promoting effect of EEKN was correlated with regulation of tyrosinase and TRP-1 protein through cAMP/PKC pathway.

• Journal of Life Science
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• v.25 no.10
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• pp.1115-1123
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• 2015

The purpose of this study was to investigate the potential of Nelumbo nucifera G. leaf (NNL) extract as a cosmetic additive. The electron-donating ability of the NNL extract at a concentration of 1,000 μg/ml was 67.83%. In xanthine oxidase, the inhibition effect of the NNL extract was 92.7% at the same concentration. For whitening effects, tyrosinase inhibition effect of NNL extract was 42.7% at a 1,000 μg/ml concentration. The cell toxicity of the NNL extract was examined in melanoma cells (B16F10) using a 3-[4, 5–dimethyl–thiazol–2–yl]-2, 5-diphenyl-tetrazoliumbromide (MTT) assay. The cell toxicity assay revealed that the NNL extract had a toxicity of 81.61% at a concentration of 1,000 μg/ml The microphthalmia-associated transcription factor (MITF), tyrosinase related protein-1 (TRP-1), tyrosinase related protein-2 (TRP-2), and tyrosinase protein expression inhibitory effect by Western blot of NNL extract were measured by a Western blot at concentrations of 25, 50, and 100 μg/ml. At a 100 μg/ml concentration of the NNL extract, the expression of the MITF, TRP-1, TRP-2, and tyrosinase protein was decreased by 69.59%, 27.74%, 67.33%, and 67.78% respectively. The MITF, TRP-1, TRP-2 and tyrosinase mRNA expression inhibitory effect were measured by reverse transcription- polymerase chain reaction (PCR) at concentrations of 25, 50, and 100 μg/ml. GAPDH was used as a positive control. At a concentration of 100 μg/ml of the NNL extract, the expression of MITF, TRP-1, TRP-2, and tyrosinase mRNA was decreased by 67.51%, 71.36%, 85.74%, and 83.64%, respectively. These findings suggest that the NNL extract has antioxidant and whitening effects and that it has great potential as a cosmetic ingredient.

• Journal of Physiology & Pathology in Korean Medicine
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• v.24 no.2
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• pp.296-301
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• 2010

The purpose of this study was to investigate the mechanism of Hexane extract of Fructus Ligustri Lucidi (HFLL)-induced regulation of melanogenesis. An apparent down-regulatory effect of tyrosinase activity was observed when B16F10 cells were incubated with HFLL. Interestingly, HFLL did not inhibit the catalytic activity of cell-free tyrosinase from B16F10 cells, whereas kojic acid directly inhibited tyrosinase activity. Regarding protein levels of melanogenic enzymes, the amounts of tyrosinase and tyrosinase-related protein 1 (TRP-1) were decreased by HFLL, while the amount of tyrosinase-related protein 2 (TRP-2) slightly was reduced after incubation with HFLL. Treatment with HFLL was found to down-regulate microphthalmia-associated transcription factor (MITF). These results suggest that HFLL is an effective inhibitor of pigmentation caused by down regulation via MITF, tyrosinase, and TRP-1 expressions.

• Journal of Life Science
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• v.27 no.3
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• pp.318-324
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• 2017

This study was performed to improve the antioxidant and skin-whitening activities of 70% ethanol extract from Sesamum indicum L. (SIL). The electron-donating ability of the SIL extract was 71.7% at a concentration of $1,000{\mu}g/ml$. The whitening effects that was measured by tyrosinase inhibition assay. As a result, SIL extract was shown 42% at $1,000{\mu}g/ml$ concentration. The cell toxicity on B16F10 melanoma cells of SIL of 70% ethanol extract showed 84.3% at $1,000{\mu}g/ml$ concentration. The microphthalmia-associated transcription factor (MITF), tyrosinase relate protein-1 (TRP-1), tyrosinase relate protein-2 (TRP-2) and Tyrosinase protein and mRNA expression inhibitory effect of SIL extract were measured by western blot and reverse transcription- polymerase chain reaction (PCR) at 50, 250, $500{\mu}g/ml$ concentration. Consequently, the MITF, TRP-1, TRP-2, Tyrosinase protein expression inhibitory effect of SIL extract was decreased by 68.3%, 39.2%, 89.7%, 22.3%, respectively, at $500{\mu}g/ml$ concentration. Moreover, MITF, TRP-1, TRP-2, Tyrosinase mRNA expression inhibitory effect by reverse-transcription-PCR of SIL extract was decreased by 81.8%, 66.5%, 84.2%, 68.1%, respectively, at $500{\mu}g/ml$ concentration. Therefore, we excellently identified the antioxidant activities and whitening effect of SIL extract, and this finding suggested that SIL extract has great potential as a cosmetic ingredients.

• Korean Journal of Food Science and Technology
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• v.51 no.1
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• pp.58-63
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• 2019

In this study, the inhibitory effects of crude polysaccharide fractions separated from Perilla frutescens Britton var. acuta Kudo (PCP) on melanin synthesis and tyrosinase activity were observed. B16F10 melanoma cells were treated with 125 and $250{\mu}g/mL$ of PCP for 24 hours. Using these optimal concentrations, inhibition of melanin synthesis inhibition was measured, and PCP treatment significantly reduced melanin synthesis induced by 3-isobutyl-1-methylxanthine (IBMX). In addition, western blotting analysis on B16F10 melanoma cells showed that PCP inhibited tyrosinase, microphthalmia-associated transcriptipn factor, tyrosinase related protein-1, and tyrosinase related protein-2 expression. Therefore, these results indicate that PCP may have potential inhibitory activity against melanin synthesis and may be a natural ingredient useful for the development of whitening materials in cosmetics and functional foods.

• Biomolecules & Therapeutics
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• v.20 no.3
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• pp.340-345
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• 2012

Sarsasapogenin (SAR) is a steroidal sapogenin that is used as starting material for the industrial synthesis of steroids. It has various pharmacological benefits, such as antitumor and antidepressant activities. Since its effect on melanin biosynthesis has not been reported, we used murine melanocyte melan-a cells to investigate whether SAR influences melanogenesis. In this study, SAR significantly increased the melanin content of the melan-a cells from 1 to 10 ${\mu}M$. Based on an enzymatic activity assay using melan-a cell lysate, SAR had no effect on tyrosinase and DOPAchrome tautomerase activities. It also did not affect the protein expression of tyrosinase-related protein 1 and DOPAchrome tautomerase. However, protein levels of tyrosinase and microphthalmia-associated transcription factor were strongly stimulated by treatment with SAR. Therefore, our reports suggest that SAR treatment may induce melanogenesis through the stimulation of tyrosinase and microphthalmia-associated transcription factor expression in melan-a cells.