• Journal of Nutrition and Health
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• v.52 no.2
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• pp.168-175
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• 2019

Purpose: Barely sprout is a well-known oriental herbal medicine with a wide range of health benefits. Recent studies have provided scientific evidence of its therapeutic effects with expanded application. This study investigated anti-melanogenic effect of barley sprout water extract (BSE) in murine melanocyte B16F10. Methods: Various concentrations (0, 50, 125, and $250{\mu}g/mL$) of BSE and arbutin (150 ppm) were applied to B16F10 stimulated with or without alpha-melanocyte stimulating hormone (100 nM) for 72 hours. The whitening potency of BSE was determined altered cellular melanin contents. Activity and expression of tyrosinase and microphthalmia-associated transcription factor (MITF) were also assayed. Results: Experimental results revealed that treatment with BSE reduced cellular melanin production by approximately 40% compared to the control. Molecular findings supported that suppressed activity and expression of tyrosinase and MITF proteins by BSE were associated with declined cellular melanogenesis. Furthermore, anti-melanogenic effect of BSE ($250{\mu}g/mL$) was similar to that of arbutin, a commonly used whitening agent. Lastly, polyphenols including p-coumaric, ferulic, and vanillic acids were identified in BSE using HPLC analyses. They might be potential active ingredients showing such melanogenesis-reducing effect. Conclusion: BSE was evident to possess favorable anti-melanogenic potency in an in vitro model. As a natural food sourced material, BSE could be an effective depigmentation agent with potential application in pharmaceutical and cosmetic industries.

• Korean Journal of Pharmacognosy
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• v.42 no.4
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• pp.323-328
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• 2011

We investigated antioxidative activity, tyrosinase inhibitory activity, and melanin production inhibitory activity of the methanol extract of Paeoniae Radix and its fractions. The total polyphenol content of the extract was 73.45 mg/g, and content of the ethyl acetate fraction was 514.50 mg/g as the highest content of fractions. In DPPH radical scavenging ability, $SC_{50}$ values of the ethyl acetate was 3.86 ${\mu}g/ml$ as a result of greater activity in the positive control (ascorbic acid). Moreover, tyrosinase inhibitory activity of the ethyl acetate fraction showed higher activity than arbutin used as a positive control. In the cytotoxicity measurement by MTT assay, the extract and fractions were exhibited cell viabilities of 76.96~157.26% against Raw 264.7 and B16F10 cell in concentration of 10~100 ${\mu}g/ml$. In nontoxic concentration range, the ethyl acetate fraction showed strong melanin production inhibitory effect in ${\alpha}$-melanocyte stimulating hormone (${\alpha}$-MSH)-stimulated B16F10 cell. As a result, the ethyl acetate fraction of the methanol extract from Paeoniae Radix could be applicable to functional materials for skin-whitening agents.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2018.10a
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• pp.95-95
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• 2018

In this study, we investigated the effect of branch extracts from Vaccinium oldhamii (VOB) on melanin synthesis in B16F10 cells. VOB promoted melanin production in absence or presence of ${\alpha}$-melanocyte-stimulating hormone (${\alpha}-MSH$) in B16F10 cells. However, VOB did not affect the expression of tyrosinase and TRP-1 associated with melanin synthesis at the mRNA and protein level in B16F10. But, VOB decreased TRP-2 protein level and induced tyrosinase activation in B16F10 cells. Inhibition of tyrosinase activity and tyrosinase knockdown attenuated VOB-mediated melanin synthesis. In conclusion, it is thought that VOB may stimulate melanin synthesis through activating tyrosinase activity.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2019.10a
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• pp.76-76
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• 2019

The purpose of this study was to evaluate the antioxidant and whitening action of Fagopyrum esculentum and F. tataricum buckwheat seeds depending on their germination time. In a previous study, we reported significant changes in sprout yield and phytochemical content in ethanol extracts from F. esculentum and F. tataricum seeds with increase in germination times. DPPH radical scavenging activities of F. tataricum increased with increasing germination time, whereas that of F. esculentum decreased. Next, we investigated anti-melanogenic activities of these species by estimation of melanin content and tyrosinase activity. Inhibition of melanin production in ${\alpha}-MSH$ (${\alpha}$-melanocyte stimulating hormone)-induced B16F10 cells by extracts from these seeds were analyzed. Among the two, F. tataricum extracts were characterized by higher inhibitory activity against melanin production. In addition, when B16F10 cells were incubated with L-DOPA for detection of in situ tyrosinase activity, F. tataricum and F. esculentum extracts were observed to reduce melanin production in cells. Taken together, these results indicate that extracts from buckwheat seeds could influence cellular processes via modulation of tyrosinase activity. Hence, buckwheat seeds could be utilized as whitening agents in the cosmetic industries and as therapeutics for hyperpigmentation disorders in a clinical setting.

• Journal of Microbiology and Biotechnology
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• v.29 no.8
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• pp.1204-1211
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• 2019

Fungal exopolysaccharides are important natural products having diverse biological functions. In this study, exopolysaccharides from Fomitopsis castanea mycelia (FEPS) were prepared, and the highest mushroom tyrosinase inhibitory activity was found. FEPS were prepared from cultivation broth by ethanol precipitation method. The extraction yield and protein concentration of FEPS were 213.1 mg/l and 0.03%, respectively. FEPS inhibited mushroom tyrosinase with the half maximal inhibitory concentration ($IC_{50}$) of 16.5 mg/ml and dose-dependently inhibited cellular tyrosinase activity (63.9% at $50{\mu}g/ml$, and 83.3% at $100{\mu}g/ml$) in the cell-free extract of SK-MEL-5 human melanoma cell and ${\alpha}$-melanocyte-stimulating hormone (${\alpha}-MSH$)-stimulated melanin formation in intact SK-MEL-5 human melanoma cell. The $IC_{50}$ of FEPS against NO production from RAW264.7 macrophage cells was $42.8{\pm}0.64{\mu}g/ml$. By in vivo study using a zebrafish model, exposure of FEPS at $400{\mu}g/ml$ to dechorionated zebrafish embryos for 18 h decreased the pigment density, compared to that without FEPS-treated control.

• 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.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.24 no.1
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• pp.25-35
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• 2011

Objective : The present study was designed to assess the potential inhibitory activity of an ethanol extract of Belamcandae Rhizoma (EBR) on the alpha-melanocyte stimulating hormone (${\alpha}$-MSH)-induced melanogenesis signal pathway in B16F10 melanoma cells. Methods : Several experiments were performed in B16F10 melanoma cells. We studied tyrosinase activity, melanin content, cell-free tyrosinase activity and DOPA stain, and performed Western blots and RT-PCR for proteins and mRNA involved in melanogenesis. Results : ${\alpha}$-MSH-induced tyrosinase activity and melanin content were inhibited significantly by EBR. EBR markedly suppressed the protein expression level of tyrosinase in B16F10 melanoma cells. On the other hand, the expression of tyrosinase-related protein-1 (TRP-1) and -2 (TRP-2; DCT) were not affected by EBR. To elucidate the mechanism of the depigmenting property of EBR, we examined the involvement EBR in cAMP response element binding (CREB) protein phosphorylation and microphthalmia-associated transcription factor (MITF) signalling induced by ${\alpha}$-MSH. EBR did not regulate CREB phosphorylation and MITF expression by ${\alpha}$-MSH. Nevertheless, the mRNA expression of tyrosinase was significantly attenuated by EBR treatment without changes in the expression of TRP-1 and -2 mRNA. Conclusion : Our study suggested that EBR inhibits ${\alpha}$-MSH-induced melanogenesis by suppressing tyrosinase mRNA.

• Journal of Physiology & Pathology in Korean Medicine
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• v.21 no.6
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• pp.1456-1461
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• 2007

Melanogenesis is induced mainly by ultraviolet radiation of sunlight and ${\alpha}-melanocyte$-stimulating hormone (${\alpha}-MSH$) which binds to a specific G protein coupled receptor. The purpose of this study was to investigate the mechanism of melanogenesis inhibition in B16/F10 cells by methanol extract of Rubus coreanus Miquel (RCM). In the present study, ${\alpha}-MSH$ and forskolin led to a stimulation of melanin synthesis that appeared to result from an increased tyrosinase activity and melanin content. However, RCM inhibited the ${\alpha}-MSH$- and forskolin-induced melanin synthesis. In addition, RCM abolished the ${\alpha}-MSH$- and forskolin-induced cytoplasmic dendricity. Regarding protein levels of the melanogenic enzymes, the amounts of tyrosinase and tyrosinase-related protein 1 (TRP-1) were increased after incubation with α-MSH and forskolin. The treatment of RCM decreased the ${\alpha}-MSH$- and forskolin-induced expression levels of tyrosinase and TRP-1. Based on these findings, it is likely that RCM exerts its depigmenting effects in B16/F10 cells through the suppression of tyrosinase and TRP-1 expression, which are key enzymes for melanogenesis.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.7
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• pp.1029-1037
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• 2012

The present study was designed to investigate the effect of ${\alpha}$-melanocyte-stimulating hormone (${\alpha}$-MSH), nitric oxide (NO) and L-cysteine on melanin production and expression of related genes MC1R, Tyr, Tyrp-1 and Tyrp-2 in muzzle melanocytes of differently colored three native Hanwoo cattle. Muzzle samples were taken from black, brindle and brown Hanwoo and purified melanocytes were cultured with ${\alpha}$-MSH, nitric oxide and L-cysteine at 100 nM, $50{\mu}M$ and 0.07 mg/ml of media respectively. The amounts of total melanin, eumelanin and mRNA expression at Tyr, Tyrp-1, Tyrp-2 and MC1R levels were quantified. ${\alpha}$-MSH and nitric oxide significantly increased (p<0.05) the amount of total melanin in black and brindle whereas eumelanin production in brown Hanwoo muzzle melanocytes. On the contrary, L-cysteine greatly (p<0.05) depressed the eumelanin production in black color but increased in brown. Simultaneously, up regulation of Tyr by nitric oxide and ${\alpha}$-MSH and down regulation of Tyr, Tyrp-2 and MC1R genes by L-cysteine were observed in muzzle melanocytes of all three phenotypes. The results of this study revealed nitric oxide and ${\alpha}$-MSH contribute hyper-pigmentation by enhancing eumelanogenesis whereas L-cysteine contributes to pheomelanin production in different colored Hanwoo muzzle melanocytes.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.14 no.1
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• pp.209-225
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• 2001

Recently many efforts were focused to understand the mechanical insights of melanogenesis to develop the agents for hyper-pigmentation and hypo-pigmentation. In the melanin biosynthetic pathway, tyrosinase is the rate limiting enzyme, and ${\alpha}$-melanocyte stimulating hormone(MSH) or cAMP-elevating agents stimulate melanogenesis and enhance the melanin synthesis and the tyrosinase activity. The author has analyzed the effects of Radix Trichosanthis on the basal melanogenic activities of B16/F10 mouse melanoma cells, and on the ${\alpha}$-MSH or forskolin-induced melanogenesis. Radix Trichosanthis alone markedly suppressed melanin content and tyrosinase activity in a dose-dependent manner. Pretreatment of the cells with Radix Trichosanthis also suppressed the increase of ${\alpha}$-MSH (10 nM) or forskolin (20${\mu}M$)-induced melanin content and tyrosinase activity. The decrease in the tyrosinase activity was paralled by a decrease in the abundance of tyrosinase protein and tyrosinase promoter activity. Pretreatment of the cells with Radix Trichosanthis also inhibited the increase of forskolin($20{\mu}M$) induced the amount of tyrosinase protein and tyrosinase promoter activity. The results of DOPA staining revealed that pretreatment of the cells with Radix Trichosanthis showed less intensity than B16 melanoma cells stimulated with ${\alpha}$- MSH or forskolin. These results suggest that Radix Trichosanthis inhibits melanogenesis and abrogates ${\alpha}-MSH and cAMP-induced melanogenesis in B16 melanoma cells.