• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.17 no.1
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• pp.82-93
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• 2004

Recently many efforts were focused to understanding 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 Bl6/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.

• Journal of Physiology & Pathology in Korean Medicine
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• v.19 no.3
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• pp.662-670
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• 2005

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 bio-synthetic pathway, tyrosinase is the rate limiting enzyme, and ${\alpha}$-melanocyte stimulating hormone(MSH) stimulates melanogenesis and enhances the melanin synthesis and the tyrosinase activity. The author has analyzed the effects of Biota Orientalis Folium on the basal melanogenic activities of B16 mouse melanoma cells, and on the ${\alpha}$-MSH or tyrosinase-induced melanogenesis. Biota Orientalis Folium alone markedly suppressed melanin content and tyrosinase activity in a dose-dependent manner. The decrease of cell propagation was observed in B16 cells treated with 200${\mu}$g/ml dose of Biota Orientalis Folium, indicating that Biota Orientalis Folium-induced depigmenting effect was caused by inhibition of melanin synthesis, not due to destruction of B16 cells. Pretreatment of the cells with Biota Orientalis Folium also suppressed the increase of ${\alpha}$-MSH (10 nM) induced melanin content and tyrosinase activity. Biota Orientalis Folium inhibited the revelation of ${\alpha}$-MSH induced tyrosinase protein and tyrosinase related protein and mRNA of tyrosinase in B16 melanoma cell. These results suggest that Biota Orientalis Folium inhibits melanogenesis and abrogates ${\alpha}$-MSH and tyrosinase-induced melanogenesis in B16 melanoma cells.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.16 no.3
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• pp.129-144
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• 2003

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 Rhizoma Bletillae on the basal melanogenic activities of B16／F10 mouse melanoma cells, and on the ${\alpha}$-MSH or forskolin-induced melanogenesis. Rhizoma Bletillae alone markedly suppressed melanin content and tyrosinase activity in a dose-dependent manner. Pretreatment of the cells with Rhizoma Bletillae also suppressed the increase of ${\alpha}$-MSH (100 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 Rhizoma Bletillae 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 Rhizoma Bletillae showed less intensity than B16 melanoma cells stimulated with ${\alpha}$-MSH or forskolin. These results suggest that Rhizoma Bletillae inhibits melanogenesis and abrogates ${\alpha}$-MSH and cAMP-induced melanogenesis in B16 melanoma cells.

• The Korea Journal of Herbology
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• v.28 no.3
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• pp.69-74
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• 2013

Objectives : Since hypopigmentation is known to increase the risk of skin cancer, melanogenesis in the skin needs to be regulated. Here, we evaluated the melanogenesis stimulatory effects of a modified Goagium herbal remedy (HR) and HR+ox bile (Bos taurus domesticus) extract (OBE) to address hypopigmentation disorders. Methods : B16F10 melanoma cells were treated with different dosages of HR and HR+OBE for 24 to 48 h after 1 h of 10 nM ${\alpha}$-melanocyte stimulating hormone (${\alpha}$-MSH). After the treatment, cell viability, tyrosinase activity, melanin synthesis and the expression of genes related to melanin synthesis were measured and the regulation of the ${\alpha}$-MSH signalling through cAMP responding element binding protein (CREB) was determined. Results : HR and HR+OBE with the ranges of $15{\sim}100{\mu}g/mL$ did not affect cell viability in melanoma cells. The 1 h treatment of HR+OBE (50 and $100{\mu}g/mL$) potentiated the phosphorylation of CREB by enhancing ${\alpha}$-MSH signaling and its 24 h treatment increased CREB expression. Consistent with CREB potentiation, their treatment for 24 h, the expression of microphthalmia-associated transcription factor (MIFT), tyrosinase, tyrosinase related protein (TRP)-1 and TRP-2 were increased in realtime PCR. Ultimately, the 48 h treatment of HR+OBE (50 and $100{\mu}g/mL$) increased tyrosniase activity and melanin contents in the melanoma cells in comparison to the control. Conclusions : HR+OBE (50 and $100{\mu}g/mL$) increases melanin synthesis in B16F10 melanoma cells via the stimulation of tyrosinase activity and expression of MIFT, tyrosinase, TRP-1 and TRP-2. HR+OBE can be used as the a possible treatment for hypopigmentation of the skin.

• Journal of the Korean Chemical Society
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• v.44 no.6
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• pp.533-540
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• 2000

Melanization of B16 melanoma cells was comparatively studied by the aqueous extract of Epimedium koreanum Nakai(EK) and $\alpha$-MSH. In addition, inhibitory effects of RA was investigated. B16 melanoma cells(about 1${\times}10_5$) have been shown an increase in tyrosinase activity and melanin contents in proportion to concentration of $\alpha$-MSH when treated with $\alpha$-MSH and incubated for 72 hrs. They indicated a 350% increase in tyrosinase activity and a 290% increase in melanin contents at 8 ng/mL. In case of EK, they have been shown a 200% increase in tyrosinase activity and a 180% increase in melanin contents at 100 ${\mu}g$/mL. In addition of RA to the above condition, they have been shown an inhibition from 350% to 210% in tyrosinase activity and from 290% to 250% in melanin contents in $\alpha$-MSH, and inhibition from 200% to 100% in tyrosinase activity and from 180% to 120% in melanin contents in EK. From the above results, it is suggested that EK promotes melanization of B16 melanoma cells through cAMP pathway, whereas RA inhibits it.

• Journal of Food Hygiene and Safety
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• v.30 no.3
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• pp.258-264
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• 2015

Withania somnifera has been used in folk medicine to treat various ailments for centuries. In this studies to investigate the whitening effect of Withania somnifera extracts as an active ingredient for whitening cosmetics, the antioxidant capacity and the effects of Withania somnifera extracts on melanogenesis in B16-F10 melanoma cells were identified. Withania somnifera extracts significantly reduced both tyrosinase activity and melanin content in a concentration-dependent manner. Furthermore, it was found that Withania somnifera extracts decreased ${\alpha}-MSH$ (melanocyte-stimulating hormone)-induced tyrosinase activity and MITF(microphthalmia associated transcription factor) protein expression. These data indicate that Withania somnifera extracts attenuate ${\alpha}$-MSH-stimulated melanin synthesis by modulating MITF expression and that they may be a useful therapeutic agent for treating hyperpigmentation and an ingredient of whitening cosmetics.

• Korean Journal of Veterinary Research
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• v.58 no.2
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• pp.65-72
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• 2018

The present study observed the effects of a green tea (Camellia sinensis) flower extract (GTFE) on melanin synthesis in B16-F10 melanoma cells. GTFE exhibited antioxidant activity on 2,2-diphenyl-1-picrylhydrazyl and inhibited mushroom tyrosinase activity in a dose-dependent manner. Furthermore, GTFE significantly diminished ${\alpha}-melanocyte$ stimulating hormone (${\alpha}-MSH$) stimulated cellular melanin content and tyrosinase activity throughout the concentration range evaluated. Based on RNA sequencing analysis, differential gene expression patterns observed in ${\alpha}-MSH$ stimulated B16-F10 melanoma cells were normalized by the addition of GTFE. In particular, the expression levels of melanoregulin and tyrosinase genes which are key regulating genes in melanin synthesis were up-regulated by 3.5 and 3 fold respectively by ${\alpha}-MSH$, and were normalized to control levels by the addition of GTFE. The results suggest that GTFE inhibits melanin synthesis in ${\alpha}-MSH$ stimulated B16-F10 melanoma cells by normalizing expression of genes that are essential for melanin synthesis. Overall, the results suggest that GTFE could be applied in the development of a whitening agent for the treatment of dermal hyperpigmentation.

• The Korea Journal of Herbology
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• v.27 no.4
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• pp.73-80
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• 2012

Objectives : Under normal condition melanin protects the skin from extracellular stimuli including ultraviolet (UV)-induced oxidative skin damages, but excess production and accumulation of melanin can induce hyperpigmentation causing esthetic problems. Therefore, in this study we tried to search for natural skin whitening materials from marine natural resources. Methods : Water and ethanol extracts of marine natural resources were prepared from Porphyra thalli (PT), Laminariae thallus (LT), Ostreae concha (OC), Sargassum thallus (ST), Undaria thallus (UT), Codium thalli (CT), Enteromorpha thalli (ET), Syngnathoides biaculeatus (SB), and Hippocampus coronatus (Hc). Their effects against UVB and ${\alpha}$-melanocyte stimulating hormone (${\alpha}$-MSH)-induced melanogenesis were investigated based on melanin formation in B16 mouse melanoma cells. The mRNA and protein expression of enzymes involved in the melanogenic process were further examined by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot analysis, respectively. Results : Water extract of Ostreae concha (OCW/E) effectively inhibited UVB and ${\alpha}$-MSH-induced melanin production in B16 melanocytes, which seemed to be mediated by inhibition of mRNA expression of tyrosinase and tyrosinase-related protein 1 (TRP-1). In another experiment, ethanol extracts from Porphyra thalli (PTE/E), Laminariae thallus (LTE/E), Sargassum thallus (STE/E), Undaria thallus (UTE/E), Codium thalli (CTE/E), Syngnathoides biaculeatus (SBE/E), and Hippocampus coronatus (HcE/E) significantly suppressed UVB and ${\alpha}$-MSH-induced melanin formation. Furthermore, ethylacetate fraction isolated form LTE/E (LTE/EEt) decreased UVB and ${\alpha}$-MSH-elevated extracellular melanin levels via inhibition of tyrosinase protein expression. Conclutions : These results suggest that marine natural resources such as Porphyra thalli, Laminariae thallus, Ostreae concha, Sargassum thallus, Undaria thallus, Codium thalli, Syngnathoides biaculeatus and Hippocampus coronatus have anti-melanogenic effects, thereby exhibiting high potentials to be utilized as one of the ingredients for the development of new whitening functional cosmetics.

• Korean Journal of Pharmacognosy
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• v.46 no.4
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• pp.315-320
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• 2015

This study was carried out to investigate the effect of ethanolic extract Salicornia herbacea (SHEE) on melanogenesis and mechanism. The SHEE on ${\alpha}$-melanocyte stimulating hormone (${\alpha}$-MSH) induces melanogenesis in B16F10 melanoma cells. The effect of SHEE remarkably decreased protein expression of tyrosinase and tyrosinase relate protein (TRP)-2 increased extracellular signal related kinase (ERK) on ${\alpha}$-MSH 100 nM induced melanogenesis on B16F10 melanoma cells at dose-dependent manner. It has been reported that the activation of ERK reduce melanin synthesis by down-regulating micro-phthalmia-associated transcription fator (MITF). These results sugggest that possibility of extracted korean medicinal herbs as a functional ingredient for whitening cosmetic formula.

• YAKHAK HOEJI
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• v.56 no.4
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• pp.254-259
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• 2012

To further access honeybee (Apis mellifera L.) venom (BV) as a cosmetic ingredient and potential external treatment for topical use, we investigated its ability to inhibit tyrosinase activity and melanin biosynthesis on melanogenesis in B16F1 melanoma cells. We found that BV increased the cell viability in B16F1 melanoma cell and BV (0.01~1 ${\mu}g/ml$) inhibited melanin synthesis in with 10 nM ${\alpha}$-melanocyte-stimulating hormone (${\alpha}$-MSH) for 48 h. In addition, we used reverse transcription-polymerase chain reaction and western blotting for me melanogenesis-related genes such as tyrosinase to examine the mechanisms underlying the inhibitory effects of BV on melanogensis. BV inhibited direct tyrosinase activity, which decreased melanin synthesis in ${\alpha}$-MSH stimulated B16F1 melanoma cells. Thease findings suggest that BV induces the downregulation of melanogenesis by inhibiting tyrosinase activation.