• Horticultural Science & Technology
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• v.31 no.6
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• pp.813-820
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• 2013

Monoterpenoids were recently found as main biologically active compounds which is responsible for various physiological effect in goat's beard (Aruncus dioicus). Ethyl acetate extract of A. dioicus (ADE) was treated to B16F10 melanoma cells for the examination of whitening activity. MTT assay was performed to evaluate cell toxicity and the result showed that slight cell toxicity (> 10%) by over $500{\mu}g{\cdot}mL^{-1}$. Thus, 0, 5, 10, or $50{\mu}g{\cdot}mL^{-1}$ ADE was used for further experiments. We found that tyrosinase activity was decreased according to ADE concentration, and the total melanin content was also dramatically reduced. Especially with $50{\mu}g{\cdot}mL^{-1}$ ADE treatment tyrosinase activity was reduced to 35.6%, and 58.8% of melanin content was lowered. In addition, whitening related proteins including tyrosinase, tyrosinase related protein 1 (TRP1), TRP2, microphthalmia associated transcription factor (MITF) and cAMP and protein kinase A (PKA) were reduced by ADE treatment. It caused decreased phosphorylation of cAMP response binding protein (CREB) but increased phosphorylation of extracellular signal related kinase (ERK). Therefore, in this paper we would like to suggest the potent usage of A. dioicus natively grown in Ulleungdo, Korea as materials of functional cosmetics by confirming whitening activity related with melanin content.

• Journal of Applied Biological Chemistry
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• v.60 no.4
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• pp.327-332
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• 2017

In this study, the whitening activity of Nymphoides indica extract in B16F10 cells were measured. Inhibition rate of tyrosinase from mushroom was 42% at $1,000{\mu}g/mL$. And inhibition of tyrosinase and melanin biosynthesis in B16F10 cells were 26 and 25% at $5{\mu}g/mL$, respectively. The expression levels of cAMP and protein kinase A (PKA), which are higher levels of melanin-related factors, were found to be decreased in a dose-dependent manner. In addition, the expression rate of protein and mRNA of tyrosinase, tyrosinase related protein 1 (TRP1), tyrosinase related protein 2 (TRP2) and microphthalmia associated transcription factor (MITF). In this study, it was confirmed that the N. indica extract effectively inhibited the activity of tyrosinase, TRP1, TRP2 and MITF as well as the activity of PKA by effectively inhibiting cAMP. Therefore, it was confirmed that the N. indica extract has high value as a functional material.

• Journal of Life Science
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• v.20 no.11
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• pp.1617-1624
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• 2010

Recently, it has been found that Asiasari radix showed a hypopigmenting effect on melanogenesis through activation of mitogen-activated protein kinase (MEK)/extracellular signal-activated kinase (ERK) in B16F10 melanoma cells. However, the hypopigmenting effect of A. radix on the $\alpha$-melanocyte stimulating hormone ($\alpha$-MSH)-stimulated melanogenesis has remained unknown. The purpose of this study was to investigate the inhibitory mechanism of the partially purified A. radix (PPAR)-induced hypopigmentating effects on $\alpha$-MSH-stimulated melanogenesis in B16F10 mouse melanoma cells. PPAR strongly inhibited tyrosinase activity and leads to decreased melanin synthesis in $\alpha$-MSH-stimulated B16F10 melanoma cells. PPAR also decreased the $\alpha$-MSH-induced over-expression of the melanogenic enzymes, tyrosinase, tyrosinase-related protein (TRP)-1, dopachrome tautomerase (Dct) and microphthalmia-associated transcription factor (MITF). We further showed that PPAR inhibits $\alpha$-MSH-induced melanogenesis via phosphorylation of MEK/ERK and PI3K/Akt, and that their activation was blocked by MEK inhibitors, PD98059 and PI3K inhibitors, LY294002 in $\alpha$-MSH-stimulated B16F10 melanoma cells. These results suggest that PPAR inhibits $\alpha$-MSH-induced melanogenesis by activation of MEK/ERK and PI3K/Akt through MITF degradation, which may lead to down-regulation of tyrosinase.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.39 no.2
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• pp.159-166
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• 2013

In order to develop new skin whitening agents, we prepared the EtOAc layer (AJE) after enzyme treatment of 75% EtOH extract of the Alnus Japonica Steud. We measured their tyrosinase inhibitory activity in vitro and melanin synthesis inhibitory activity in B16-F1 melanoma cells. They did not show inhibitory activity against mushroom tyrosinase but showed melanin synthesis inhibitory activity in a dose-dependent manner. In a melanin synthesis inhibition assay, AJE suppressed melanin production up to 52% at a concentration of $40{\mu}g/mL$. To elucidate the mechanism of the inhibitory effects of AJE on melanogenesis, we measured expression of melanogenesis-related proteins by the western blot assay. As a result, AJE suppressed the expression of tyrosinase related protein 1 (TRP-1) and microphthalmia associated transcription factor (MITF). Moreover, AJE increased the expression of phosphorylated extracellular signal-regulated kinase (p-ERK). These results conclude that ERK activation by AJE reduces melanin synthesis via MITF downregulation and is subsequent to the inhibition of TRP-1 expression. Therefore, we suggest that AJE could be used as active ingredients for skin whitening.

• Journal of Life Science
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• v.23 no.12
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• pp.1445-1453
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• 2013

Human skin is constantly exposed to ultraviolet (UV) radiation, polluted air, and chemical products. UV rays, in particular, will affect the skin in a variety of ways, including causing wrinkles, fine lines, rough skin, and xeroderma, thereby resulting in skin aging. This study aimed to investigate the whitening effects of Juniperus rigida Sieb., which is a cedar tree that is found throughout the world. The whitening efficacy that was measured by tyrosinase inhibition revealed 49.4% efficacy in water extract and 80.0% efficacy in ethanol extract. Among the B16F10 black cells, the effect of the ethanol extract was higher than the effect of the water extract in the restrain creation of melanin pigment, tyrosinase, microphthalmia-associated transcription factor (MITF), tyrosinase related protein-1 (TRP-1), and tyrosinase related protein-2 (TRP-2). Thus, the results of these studies demonstrated that the ethanol extract had greater efficacy than the water extract and Juniperus rigida Sieb. Ethanol extracts could be utilized as materials for functional cosmetics, such as whitening products.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.41 no.1
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• pp.21-26
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• 2015

The purpose of this study is to identify the whitening effect of Androsace umbellata extract. To discover the anti-pigmentation effective, we investigated Androsace umbellata extract on tyrosinase and melanogenesis inhibition. As results, it reduced tyrosinase activity and melanin contents in B16F1 melanoma cells in a dose-dependent manner with decreased to about 32% at a concentration of $25{\mu}g/mL$. To reveal how it works in inner-cellular level, we performed Western blot method. We found out that it also inhibited the protein expression in tyrosinase, tyrosinase related protein 1 (TRP-1), and microphthalmia associated transcription factor (MITF) in melanocytes. Therefore, we successfully identified the whitening effect of Androsace umbellata extract, and this finding suggested that Androsace umbellata extract is a considerable potent cosmetics ingredient for whitening. Based on this, we anticipated further researches about Androsace umbellata extract for gene levels and additional mechanisms to develop not only for functional cosmetics but for medicines or healthcare food.

• Journal of Life Science
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• v.31 no.11
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• pp.1019-1027
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• 2021

There are a lot of efforts to develop new compounds having skin whitening effect from natural products. Sparassis crispa is a medicinal mushroom containing more than 40% β-glucan, which exhibits anticancer and immunostimulating effects. The aim of this study was to assess the availability of β-glucan extracted from cauliflower mushroom S. crispa as a skin whitener through the evaluation of inhibitory effects of melanin synthesis and tyrosinase activity and their mechanisms. B16F1 cells were treated with S. crispa β-glucan (10, 100, and 1,000 ㎍/ml, respectively) and α-melanocyte stimulating hormone (α-MSH), simultaneously. Content of melanin synthesis and tyrosinase activity were determined. The expressions levels of tyrosinase, tyrosinase related protein-1 (TRP-1), TRP-2 and microphthalmia-associated transcription factor (MITF) were also measured by western blotting. Treatment with 10, 100 and 1,000 ㎍/ml S. crispa β-glucan and 200 nM α-MSH significantly decreased melanin synthesis by 13.9%, 18.7% and 39.5%, respectively, and tyrosinase activity by 15.6%, 26.9% and 43.2%, respectively, compared to the α-MSH alone group. In addition, S. crispa β-glucan inhibited expressions of tyrosinase, TRP-1, TRP-2 and MITF induced by α-MSH. These results indicated that S. crispa β-glucan inhibited MITF expression, thereby reducing tyrosinase expression and inhibiting melanin production in B16F1 melanoma cells. Therefore, S. crispa β-glucan might be available as a skin whitener.

• 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.

• 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.

• The Korea Journal of Herbology
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• v.29 no.6
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• pp.157-163
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• 2014

Objectives : Gamisoyo-san complex prescription were made with Angelicae Gigantis Radix, Paeoniae Radix, Atractylodes rhizome white, Hoelen, Bupleuri Radix, Moutan Cortex Radicis, Gardeniae Fructus, Zingiberis Rhizoma Crudus, Menthae Herba. The purpose of this study was to research the whitening effect of the extract from Gamisoyo-san, which is one of the used herbal complex prescription. Methods : This study investigated inhibitory effect of Gamisoyo-san in tyrosinase activity. Cell viability were performed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Then, Gamisoyo-san measured reversed-transcription-PCR for mRNA expression using B16F10 mouse melanoma cells. Results : For whitening effects, the tyrosinase inhibition effect of extract was shown to 52.4% at $5,000{\mu}g/m{\ell}$ concentration. The cell viability on B16F10 melanoma cells of Gamisoyo-san extract showed higher than 75% at $1,000{\mu}g/m{\ell}$ concentration. In this study, an experiment was performed by setting the non-toxic concentration range of 50, 150, $250{\mu}g/m{\ell}$. The Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as a positive control. The microphthalmia-associated transcription factor (MITF), tyrosinase related protein-1 (TRP-1), tyrosinase related protein-2 (TRP-2), tyrosinase mRNA expression inhibitory by reverse transcription-PCR of Gamisoyo-san extract were decreased by 95.3%, 98.8%, 96.3% and 49.5% at $250{\mu}g/m{\ell}$ which the highest concentration. Conclusions : All these findings could verify that whitening effects of Gamisoyo-san extract by tyrosinase inhibitory activity and mRNA expression. The Gamisoyo-san could be used as material for functional cosmetics, such as skin whitening products.