• Fisheries and Aquatic Sciences
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• v.21 no.11
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• pp.35.1-35.7
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• 2018

Background: The aim of this study was to investigate the in vitro inhibitory effects of Fucofuroeckol-A isolated from Eisenia bicyclis against tyrosinase activity and 3-isobutyl-1-methylxanthine (IBMX)-induced melanin biosynthesis in B16F10 melanoma cells. Result: Among the ethanolic (EtOH) extract of E. bicyclis and its organic solvent fractions, the ethyl acetate (EtOAc) soluble fraction showed a noticeable inhibitory effect on mushroom tyrosinase with an $IC_{50}$ value of $37.6{\pm}0.1{\mu}g/mL$. Repeated column chromatography of the active EtOAc fraction resulted in the isolation of Fucofuroeckol-A. It evidenced more potent tyrosinase inhibitory effect with an $IC_{50}$ value of $11.4{\pm}1.4{\mu}M$ than arbutin ($IC_{50}=1076.6{\pm}44.3{\mu}M$), which was used as a positive control. Lineweaver-Burk plots suggest that Fucofuroeckol-A plays as a noncompetitive inhibitor against tyrosinase. Furthermore, we have evaluated the inhibitory effects of Fucofuroeckol-A on IBMX-induced melanin formation in B16F10 melanoma cells. Fucofuroeckol-A ($12.5-100{\mu}M$) exhibited a significant inhibition of melanin production in the melanoma cells. Conclusion: In the present study, we suggested that Fucofuroeckol-A might prove possibility as a novel inhibitor of melanin biosynthesis in cosmetic applications.

• Korean Journal of Plant Resources
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• v.34 no.6
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• pp.510-516
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• 2021

Whitening agents derived from natural sources which do not have side effects are sought after. Schizophragma hydrangeoides is an edible plant that grows wild on Jeju Island. We aimed to determine whether S. hydrangeoides extracts show anti-melanogenic activity. Here, we found that 70% ethanol extracts of S. hydrangeoides leaf suppressed α-melanocyte-stimulating hormone-induced melanogenesis in B16F10 mouse melanoma cells. This activity of anti-melanogenesis in B16F10 cells were investigated by determining melanin content and tyrosinase activity, and by performing western blotting. The 70% ethanol extract downregulated tyrosinase and tyrosinase-related protein 1. In addition, the n-hexane fraction of S. hydrangeoides leaf (HFSH) exhibited significant anti-melanogenic activity among the various solvent fractions tested without reducing the viability of B16F10 cells. Taken together, these results indicate that extracts from S. hydrangeoides leaf can influence cellular processes via modulation of tyrosinase activity. Hence, S. hydrangeoides can be used as a whitening agent in the cosmetic industry and as a therapeutic agent for treating hyperpigmentation disorders in the clinic.

• 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 Life Science
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• v.29 no.9
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• pp.972-976
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• 2019

Polyopes affinis is a kind of red algae found in the South coast and near Jeju Island of Korea. The purpose of this study was to investigate the effects of Polyopes affinis ethanol extract (PAEE) on melanogenesis in ${\alpha}-MSH$ stimulated B16F10 melanoma cells. Melanoma cells were cultured for 72 hr treated with PAEE. Total melanin content and the activity of tyrosinase, a key enzyme in melanogenesis, were measured. When the melanin content in B16F10 melanoma cells was tested, PAEE was decreased in a dose-dependent manner: treatment with 100, 300, and $500{\mu}g/ml$ caused 25%, 30%, and 35% reduction, respectively. Treatment of 100, 300, and $500{\mu}g/ml$ of PAEE caused 6%, 12%, and 21% reduction of tyrosinase activities in B16F10 melanoma cells. Also, PAEE suppressed the expression of tyrosinase, tyrosinase-related protein-1, tyrosinase-related protein-2, and melanocyte-inducing transcription factor in B16F10 melanoma cells. A concentration of $500{\mu}g/ml$ of PAEE showed a greater decrease in tyrosinase activity, melanin content, and melanogenic enzyme protein expression. These results indicate that PAEE inhibits melanin synthesis and tyrosinase activity, and Polyopes affinis ethanol extract could be used as a functional whitening agent.

• The Korea Journal of Herbology
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• v.34 no.2
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• pp.75-82
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• 2019

Objectives : In this study, various biological effects of Curcuma longa L. have been studied, however, beneficial effect of Curcuma longa L. in skin health remain still unclear. In this study, Curcuma longa L. water extract (CLE) was prepared. Inhibitory effect of CLE on melanin accumulation of B16F10 cells and expression levels of skin fibril-related proteins of human skin fibroblasts (HSF) were evaluated. Methods : The cytotoxic effect of CLE in B16F10 cells and HSF were examined by MTT assay. Inhibitory effect of CLE on the ${\alpha}-MSH-$ and IBMX-induced melanin accumulation and tyrosinase activity were evaluated in B16F10 cells. The expression levels of connective tissue growth factor (CCN2), Smad2, procollagen $1{\alpha}2$, collagen $1{\alpha}2$, and fibronectin in CLE-treated HSF were analyzed by western blotting. Results : The CLE treatment (concentrations 10 to $400{\mu}g/ml$) for 72 h did not affect to the B16F10 viability. However, 200 and $400{\mu}g/ml$ of CLE treatment for 24 h showed cytotoxic effect in HSF. Therefore, the concentrations 10, 50, and $100{\mu}g/ml$ of CLE were chosen in this study. The CLE treatment for 72 h dose dependently and significantly suppressed melanin accumulation and tyrosinase activity of B16F10 cells. In addition, the CLE treatment up-regulated expression levels of skin fibril-related proteins such as CCN2, Smad2, procollagen $1{\alpha}2$, collagen $1{\alpha}2$, and fibronectin. Conclusions : In conclusion, these results suggest that the CLE could be used as a natural material for skin health.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.1
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• pp.167-174
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• 2014

Tumor angiogenesis, growth and metastasis are three closely related processes. We therefore investigated the effects of barbigerone on all three in the B16F10 tumor model established in both zebrafish and mouse models, and explored underlying molecular mechanisms. In vitro, barbigerone inhibited B16F10 cell proliferation, survival, migration and invasion and suppressed human umbilical vascular endothelial cell migration, invasion and tube formation in concentration-dependent manners. In the transgenic zebrafish model, treatment with $10{\mu}M$ barbigerone remarkably inhibited angiogenesis and tumor-associated angiogenesis by reducing blood vessel development more than 90%. In vivo, barbigerone significantly suppressed angiogenesis as measured by H and E staining of matrigel plugs and CD31 staining of B16F10 melanoma tumors in C57BL/6 mice. Furthermore, it exhibited highly potent activity at inhibiting tumor growth and metastasis to the lung of B16F10 melanoma cells injected into C57BL/6 mice. Western blotting revealed that barbigerone inhibited phosphorylation of AKT, FAK and MAPK family members, including ERK, JNK, and p38 MAPKs, in B16F10 cells mainly through the MEK3/6/p38 MAPK signaling pathway. These findings suggested for the first time that barbigerone could inhibit tumor-angiogenesis, tumor growth and lung metastasis via downregulation of the MEK3/6/p38 MAPK signaling pathway. The findings support further investigation of barbigerone as a potential anti-cancer drug.

• YAKHAK HOEJI
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• v.51 no.5
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• pp.350-354
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• 2007

Taurine has been shown to be tissue-protective against oxidant-induced injury and is a powerful regulator of the immune system. However, there is no study on the antimelanogenic effect of taurine. In this study, we investigated the whitening effect of taurine in B16F10 mouse melanoma cells. Cell viability was measured by MTT assay. We examined melanin contents and tyrosinase activity according to time and concentration. Extracellular signal regulated kinase (ERK) is an important regulator of melanogenesis. It has been reported that activated ERK induced microphthalmia associated transcription factor (MITF) phosphorylation and its subsequent degradation and thus reduced melanin synthesis. In our B16F10 cell culture system, taurine led to decrease melanin contents by 21% at 48 hr. We then observed taurine effects on ERK-P, MITF and tyrosinase by Western blot. ERK was activated at 18 hr and 24 hr, whereas MITF reduced. We could not observe any differences in the levels of tyrosinase. These results suggested that taurine inhibited melanogenesis by ERK signal pathway via MITF degradation. We expect that taurine has potential skin whitening agents in cosmetics.

• YAKHAK HOEJI
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• v.46 no.2
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• pp.137-142
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• 2002

Caesalpinia sappan L. has long been commonly used as emmenagogue, analgesic, and a cure for contusion and sprain as well as a remedy for thrombosis in the Oriental medicine. The main constituent of C. sappan is brazilein, which is an antioxidative substance that has a flavonoid structure. In this study, we examined the effect of butanol extract of C. sappan on proliferation and melanogenesis in B16/F10 melanoma cells. After 48h treatment of cells with various concentrations of butanol extract, the cells exhibited a dose-dependent inhibition in their proliferation without apotosis. Therefore, the growth retardation by the extract may be due to the cell arrest, not due to the cell death induced by cytotoxicity. We also estimated total melanin contents as a final product and activity of tyrosinase, a key enzyme, in melanogenesis of B16/F10 melanoma cells. Our result showed that the melanin contents and tyrosinase activity were decreased in butanol extract-treated cells in a dose dependent manner compared to control group. In conclusion, it was observed that butanol extract of C. sappan inhibited melanization of these cells and therefore butanol extract could be developed as skin whitening components of cosmetics.

• YAKHAK HOEJI
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• v.49 no.1
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• pp.86-91
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• 2005

Flavonoids are class of polyphenolic compounds widely distributed in the plant kingdom, which display a variety of biological activities, including antiviral, antithrombotic, antiinflammatory, antihistaminic, antioxidant and free-radica 1 scavenging abilities. The antioxidant enzyme (AOE) system plays an important role in the defense against oxidative stress insults. To determine whether flavonoid, myricetin can exert antioxidative effects not only directly by modulating the AOE system but also scavenging free radical, we investigated the influence of the flavonoid myricetin on cell viability, different antioxidant enzyme activities, ROS level and the expression of different antioxidant emzyme in B16F10 murine melanoma cells. Myricetin in a concentration range from 6.25 to $50\;{\mu}M$ decreased superoxide dismutase (SOD) and glutathione peroxidase (GPx) enzyme activities, but catalase (CAT) activity was increased. In the myricetin-treated group, ROS levels were decreased dose-dependently. Antioxidant enzyme expression was measured by RT-PCR. Myricetin treatment of B16F10 cells increased catalase expression. Expression levels of copper zinc superoxide dismutase (CuZn SOD) were not affected by exposure of myricetin. Manganese superoxide dismutase (Mn SOD) and GPx expression levels decreased slightly after myricetin treatment. In conclusion, the antioxidant capacity of myricetin was due to CAT and free-radical scavenging.

• YAKHAK HOEJI
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• v.47 no.6
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• pp.432-437
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• 2003

It has been known that quercetin, a bioflavonoid widely distributed in fruits and vegetables as dietary-derived flavonoid and exert significant multiple biological effects such as antioxidant and anti-inflammatory, anti-tumor effects. In addition, it has been shown to have a chemoprotective role in cancer, though complex effects on signal transduction involved in cell proliferation and angiogenesis. The present study investigated whether quercetin can enhance antioxidant enzyme activity (glutathione peroxidase: GPx, superoxide dismutase: SOD, catalase: CAT) and regulate the reactive oxygen species (ROS) generation in the presence of vitamin E, L-ascorbic acid, reduced glutathione (GSH) on B16F10 murine melanoma cells. After 48h treatment of cells with quercetin in the presence of vitamin E, L-ascorbic acid, GSH, we measured the cytotoxicities by MTT assay. The cells exhibited a dose-dependent inhibition in their proliferation in the presence of vitamin E, L-ascorbic acid, GSH respectively. We also investigated the effects of antioxidant enzyme activity and ROS generation. The antioxidant enzyme activity of quercetin in the presence of vitamin E was stronger than GSH, L-ascorbic acid, the same treatments decreased ROS generation in B16F10 murine melanoma cells. Taken together, these result demonstrate that the antioxidant effect of quercetin can enhanced in the presence of vitamin E and it might plays an important role in anti-oxidative effects.