• Journal of the Korean Society of Food Science and Nutrition
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• v.42 no.3
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• pp.355-362
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• 2013

This study was performed to assess the antioxidant activities and whitening effects of protopectinase enzymes and mechanical maceration from soybeans on melanin synthesis. The whitening effects of enzyme treatment and mechanical maceration were examined by an in vitro mushroom tyrosinase assay and by assessing markers in B16BL6 melanoma cells. We assessed inhibitory effects on the expression of melanogenic enzymes, including tyrosinase, tyrosinase-related protein 1 (TRP-1), and tyrosinase-related protein 2 (TRP-2) in B16BL6 cells. Inhibitory effects on free radical generation were determined by measuring DPPH and hydroxyl radical scavenging activities. In DPPH radical scavenging activity, enzyme treatment and mechanical maceration had a potent anti-oxidant activity in a dose-dependent manner and significantly inhibited tyrosinase activity in vitro and in B16BL6 melanoma cells. There was also an inhibition in the expression of tyrosinase, TRP-1, and TRP-2 in B16BL6 melanoma cells. Our results show that soybean protopectinase treatment inhibits melanogenesis, with the underlying mechanism possibly due to the inhibition of tyrosinase activity and tyrosinase, TRP-1, and TRP-2 expression. We suggest that soybean protopectinase should be contained as natural active ingredients for antioxidant and whitening cosmetics.

• Korean Journal of Plant Resources
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• v.35 no.6
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• pp.704-712
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• 2022

Research on whitening materials using natural alternatives is actively being conducted. The aim of this study was to investigate the in vitro inhibitory effects of Ranunculus chinensis Bunge (RCB) on melanogenesis and associated enzymes, such as tyrosinase, tyrosinase-related protein (TRP)-1, and TRP-2 in B16F10 murine melanoma cells. We found that RCB extract significantly attenuated melanin synthesis and reduced the activity of intracellular tyrosinase, a rate-limiting melanogenic enzyme. Western blot analysis showed that RCB extract decreased the protein expression of tyrosinase and TRP-1. In addition, it significantly decreased the expression of microphthalmia-associated transcription factor (MITF), a key regulator of melanogenesis. Extracellular signal-regulated kinase (ERK) activation has been reported to be involved in the inhibition of melanogenesis. Thus, we investigated whether the hypopigmentary effects of RCB extract were related to the activation of ERK. RCB extract induced ERK phosphorylation in a dose-dependent manner. Furthermore, it markedly inhibited body pigmentation in a zebrafish model. Our results suggest that RCB extract inhibits melanogenesis by activating ERK pathway-mediated suppression of MITF and its downstream target genes, including tyrosinase. Therefore, RCB extract can be used as a whitening agent in the development of functional cosmetics.

• 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.

• 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 Physiology & Pathology in Korean Medicine
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• v.28 no.5
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• pp.506-511
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• 2014

Commelina communis Ledeb is a widely used medication for the treatment of antidiabetic, antioxidant and hypoglycemic agent in Korea. Alpha-melanocyte stimulating hormone (${\alpha}$-MSH) is a major factor to stimulate melanin synthesis in the skin. The purposes of this study was to investigate the inhibitory effects of extract from Commelina communis Ledeb (ECC) on ${\alpha}$-MSH-stimulated melanogenesis in B16F10 cells. ECC suppressed melanin synthesis and intracellular tyrosinase activity in B16F10 cells or ${\alpha}$-MSH-induced B16F10 cells in a dose dependent manner. In study on the melanogenic protein expressions, it had especially influence on expressions of tyrosinase and tyrosinase-related protein (TRP-1). Tyrosinase and TRP-1 expressions were gradually decreased in a dose-dependent. Additionally, the extract also decreased the ${\alpha}$-MSH-induced over-expression of tyrosinase and TRP-1. This results show that the anti-melanogenic activity of ECC is correlated with the suppression of tyrosinase and TRP-1 protein expressions in B16F10 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.

• 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.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.41 no.3
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• pp.263-268
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• 2015

The purpose of this study was to investigate anti-melanogenesis effects of mulberry seed extracts (MSE). MSE inhibited melanogenesis in melan-a cells at $10{\mu}g/mL$ without cytotoxicity. Also, MSE decreased tyrosinase, tyrosinase-related protein-1 (TRP-1) protein expression in the melan-a cells. To identify the signaling pathway of MSE, the ability of MSE to influence extracellular signal-regulated protein kinase (ERK) activation was investigated. MSE induced ERK protein expression in a dose-dependent manner. In addition, MSE presented inhibition of the body pigmentation in vivo zebrafish model. These results suggest that MSE may be an effective anti-melanogenesis agent regulating the expression of ERK protein and melanogenic enzymes.

• Journal of Life Science
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• v.27 no.4
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• pp.423-429
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• 2017

Hair color is determined by kind and amount of melanin. Melanocyte mainly synthesizes melanin from L-tyrosine by stimulation of ultra violet. Reactive oxygen species (ROS) play an important role in greying hair. Polygonum multiflorum radix has been reported to inhibit the aging process that black color of hair is turned into grey color. The aim of this study is to investigate the effect of Polygoni multiflorium radix ethanol extract (PMEE) on melanin synthesis related to black hair growth. In anti-oxidant experiment, PMEE decreased DPPH radical and increased reducing power, indicating that PMEE could eliminate ROS involved in greying hair. PMEE decreased cell viability in a dose-dependent manner. Furthermore, the effect of PMEE on the production of melanin was determined by DOPA assay and tyrosinase activity. PMEE increased tyrosinase activity and promoted melanin synthesis. In addition, the expression levels of tyrosinase, tyrosinase related protein-1 (TRP-1), tyrosinase related protein-2 (TRP-2) and microphthalmia-associated transcription factor (MITF), as well as anti-oxidant enzymes such as superoxide dismutase (SOD-3) and catalase were examined using western blot analysis. The expression levels of SOD-3 and catalase were decreased due to the enhanced antioxidant activity of PMEE. In particular, PMEE increased the expression levels of tyrosinase and TRP-2. These results suggest that PMEE could promote melanin synthesis that involved in tuning gray hair into black hair.

• 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.