• 한방안이비인후피부과학회지
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• 제26권1호
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• pp.1-18
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• 2013

Objective: Recently the demands for the effective and safe depigmentative and anti-aging agents of the skin have increased due to the medical, pharmaceutical and cosmetic reasons. The purpose of this study is to investigate the MKG(Methanol Extract of Kaempferia galanga) and their dermal bioactivity properties related to cosmeceuticals such as depigmentation. Methods: We assessed inhibitory effects of MKG on melanin production in B16/F10 melanoma cells, on mushroom tyrosinase activity, effects of MKG on the expression tyrosinase, TRP-1, TRP-2, GSK-$3{\beta}$, CREB, MITF in B16/F10 melanoma cells without cytotoxicity range. Cell viability was measured by MTT assay and tyrosinase activity was assessed using by DOPA staining, western-blot analysis. We measured inhibition of melanin synthesis and tyrosinase activity by down-regulation of melanogenic enzyme expressions in ${\alpha}$-MSH induced melanogenesis B16/F10 melanoma cells. Results: MKG inhibited tyrosinase-activity, total melanin contents and dendrite out-growth. MKG inhibited melanogenesis by down-regulation of tyorsinase, TRP-1, TRP-2, CREB, and MITF in B16/F10 cells. The treatment with MKG at the 12.5, $25{\mu}g/ml$ level significantly inhibited the melanin synthesis induced ${\alpha}$-MSH in B16/F10 melanoma cells compared with untreated control. Conclusion: These results suggest that MKG inhibit melanin biosynthesis which is involved in hyper-pigmentation. So MKG is considered to be used as a whitening components reducing cytotoxicity.

• 대한화장품학회지
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• 제39권2호
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• pp.159-166
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• 2013

본 연구에서는 새로운 미백소재를 개발하기 위해 적양 에탄올추출물을 효소처리 후 EtOAc 분획물(AJE)을 준비하여 in vitro 상에서 이들의 tyrosinase 저해활성과 세포 수준에서의 멜라닌 합성 저해효과를 측정하였다. AJE는 mushroom tyrosinase의 활성에는 영향을 미치지 않았으나 B16-F1 melanoma cell을 이용한 멜라닌 합성 저해효과에 있어서 농도 의존적으로 멜라닌 합성을 저해하여, $40{\mu}g/mL$의 농도에서 52% 이상의 저해효과를 나타내었다. 이러한 멜라닌 합성 저해효과에 대한 작용 기전을 확인하기 위해 western blot을 통해 멜라닌 합성 경로에 관련된 단백질의 발현을 측정하였다. 그 결과 멜라닌 합성에 관여하는 효소인 tyrosinase related protein 1 (TRP-1)의 발현을 억제하였고, 이를 조절하는 전사인자인 microphthalmia associated transcription factor (MITF) 발현 역시 효과적으로 억제하였다. 또한 extracellular signal-regulated kinase (ERK) pathway를 활성화시킴으로써 phosphorylated extracellular signal-regulated kinase (p-ERK)의 발현을 상당히 증가시키는 것을 확인할 수 있었다. 이러한 결과는 AJE가 멜라닌 합성의 신호전달 경로 중 ERK pathway의 활성화를 통해 MITF의 분해를 촉진시키고 이로 인해 MITF의 발현을 감소시키며, 그 결과 멜라닌 합성에 관여하는 효소 중 TRP-1의 발현을 감소시킴으로써 멜라닌 합성을 저해하는 것으로 사료되며, 따라서 AJE는 미백용도의 기능성 원료로서의 가능성이 큰 것으로 판단된다.

• 생명과학회지
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• 제23권12호
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• pp.1495-1500
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• 2013

본 연구에서는 톳 분획물의 미백효과 및 그에 따른 생화학적 기전의 해석을 위하여 톳 분획물이 ${\alpha}$-MSH에 의해 인위적으로 유발된 melanogenesis 과정에 있어서 어떠한 영향을 미치는 지를 조사하였고, 이때 MITF, TRP1, TRP2, tyrosinase 등과 같은 melanin 생성에 관여하는 유전자들의 발현에 어떠한 변화가 유발되었는지를 조사하였다. 톳 분획물 처리가 B16F10 mouse melanoma cell의 멜라닌의 생성량과 tyrosinase의 활성에 미치는 영향을 확인한 결과, butanol fraction (BFHF) 처리군과 water fraction (WFHF) 처리군을 제외한 ethyl extract (EEHF) 처리군, dichlorimethane fraction (CFHF ) 처리군, ethyl acetate fraction (EAFHF) 처리군 모두 농도의존적으로 멜라닌 생성량과 tyrosinase 활성이 억제되었으며, MITF, TRP1, TRP2, tyrosinase의 발현이 단백질 수준에서 감소하였음을 확인하였다. 그 중 특히 EEHF 처리군과 CFHF 처리군은 50 ${\mu}g/ml$의 농도에서 멜라닌 생성량을 40.5%, 33.2% 감소시켰으며, tyrosinase 활성도 50 ${\mu}g/ml$의 농도에서 각각 53.3%, 54.1% 감소시켰다. 또한 EEHF, CFHF, EAFHF 처리군에서 MITF, TRP1, TRP2, tyrosinase의 발현이 농도 의존적으로 감소하였음을 확인하였다. 이상의 결과를 살펴볼 때 톳 분획물 중 특히 EEHF 처리군과 CFHF 처리군이 B16F10 mouse melanoma cell에서 melanin 생성에 관여하는 유전자인 MITF, TRP1, TRP2, tyrosinase 발현을 억제하므로 향후 미백 기능성 소재로서의 활용이 가능할 것이라고 사료된다.

• 동의생리병리학회지
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• 제18권3호
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• pp.830-836
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• 2004

The unique biochemical pathways in melanocytes responsible for melanogenesis provide us with a rational mechanism-based means for developing both pharmacological regulators of pigmentation and cytotoxic chemotherapeutic drugs for melanoma, Myricetin is a polyphenolic antioxidant and a component from Biotae Semen, Biotae orientalis Folium. Therefore, the present study was conducted to evaluate the effects of myricetin on the melanogenesis in human melanoma (HM₃KO) cells. The cells were treated for 5 days with myricetin at several concentrations (10 - 100 μg/ml). Treatment with myricetin dose-dependently suppressed cell growth in HM₃KO cells, But melanin formation was markedly increased by myricetin as a dose dependent manner. Myricetin did not affect to tyrosinase activity, which is a key enzyme for melanogenesis, but significantly increased the level of tyrosinase protein expression, These results suggest that myricetin stimulate melanin synthesis of human melanoma cells through the modification of tyrosinase protein expression.

• Archives of Pharmacal Research
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• 제27권3호
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• pp.334-339
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• 2004

In this study, the effects of (-)-epigallocatechin-3-gallate (EGCG) and/or hinokitiol (${\beta}-thujaplicin$) on melanogenesis were investigated. Our results showed that both EGCG and hinokitiol significantly inhibited melanin synthesis in a concentration-dependent manner, and that their hypopigmenting effects were stronger than that of kojic acid, which is known to inhibit melanin formation in melanocytes and melanoma cells. Interestingly, EGCG did not show any additive hypopigmenting effect in combination with kojic acid, though EGCG did show a synergistic effect in combination with hinokitiol. Several reports indicate that the activation of extracellular signal-regulated kinase (ERK) induces microphthalmia-associated transcription factor (MITF) degradation. Accordingly, the effects of EGCG and hinokitiol on the ERK signaling pathway were examined. EGCG and hinokitiol induced neither ERK activation nor MITF degradation. On the other hand, both EGCG and hinokitiol reduced the protein levels of MITF and of tyrosinase, the rate limiting melanogenic enzyme, whereas kojic acid had no effect. In addition, hinokitiol strongly downregulated the activity of tyrosinase, whereas EGCG or kojic acid had only a little effect. These results show that both EGCG and hinokitiol reduce MITF production, and suggest that reduced tyrosinase activity by hinokitiol explains their synergistic effect on melanogenesis.

• BMB Reports
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• 제43권12호
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• pp.824-829
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• 2010

Melanin synthesis is regulated by melanocyte specific enzymes and related transcription factors. $\beta$-carboline alkaloids including harmaline and harmalol are widely distributed in the environment including several plant families and alcoholic beverages. Presently, melanin content and tyrosinase activity were increased in melanoma cells by harmaline and harmalol in concentration- and time-dependent manners. Increased protein levels of tyrosinase, tyrosinase-related protein-1 (TRP-1), and TRP-2 were also evident. In addition, immunofluorescence and Western blot analyses revealed harmaline and harmalol increased cAMP response element binding protein phosphorylation and microphthalmia-associated transcription factor expression. In addition to studying the signaling that leads to melanogenesis, roles of the p38 MAPK pathways by the harmaline and harmalol were investigated. Harmaline and harmalol induced time-dependent phosphorylation of p38 MAPK. Harmaline and harmalol stimulated melanin synthesis and tyrosinase activity, as well as expression of tyrosinase and TRP-1 and TRP-2 indicating that these harmaline and harmalol induce melanogenesis through p38 MAPK signaling.

• Biomolecules & Therapeutics
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• 제22권1호
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• pp.35-40
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• 2014

Resveratrol is a polyphenolic compound found in various natural products such as grapes and berries and possesses anti-cancer, anti-hyperlipidemia, and anti-aging properties. Recently, it has been reported that resveratrol inhibits ${\alpha}$-melanocyte-stimulating hormone signaling, viability, and migration in melanoma cells. However, these effects have not been confirmed in vivo, specifically brownish guinea pigs. To evaluate the potential of resveratrol as a regulator of melanin for hyperpigmentation therapy, the influence of resveratrol on pigmentation was investigated by ultraviolet B-induced hyperpigmentation in brownish guinea pig skin. We found that resveratrol reduced the expression of melanogenesis-related proteins tyrosinase, tyrosinase-related proteins 1 and 2, and microphthalmia-associated transcription factor in melanoma cells. Furthermore, topical application of resveratrol was demonstrated to significantly decrease hyperpigmentation on ultraviolet B-stimulated guinea pig skin in vivo. Based on our histological data, resveratrol inhibits melanin synthesis via a reduction in tyrosinase-related protein 2 among the melanogenic enzymes. This study is the first to provide evidence supporting resveratrol as a depigmentation agent, along with further clinical investigation of resveratrol in ultraviolet B-induced skin disorders such as hyperpigmentation and skin photoaging.

• Korean Journal of Acupuncture
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• 제31권4호
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• pp.188-194
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• 2014

Objectives : In this study, we investigated the effect of ethanol extract from Rosa rugosa Thunb. flowers(ERF) on the activities of antioxidant, antiwrinkle and whitening. Methods : We measured antioxidant efficacy of ERF by using 1,1-diphenyl-2-picrylhydrazyl(DPPH) assay. Also we confirmed the inhibitory effect of ERF on collagenase activity and melanin synthesis by using collagenase assay kit and dihydroxiphenylalanine staining, respectively. To evaluate the anti-inflammatory effects of ERF, we examined the inflammatory mediator IL-6. Results : ERF showed highly efficacy in DPPH radical scavenging activity. ERF dose-dependently suppressed collagenase activity. Ultraviolet-induced production of IL-6 decreased by ERF treatment. In B16F10 cell, ERF significantly reduced tyrosinase activity and melanin synthesis. Conclusions : From the above results, it was indicated that ERF could be utilized as anti-aging and whitening cosmetic ingredients.

• Biomolecules & Therapeutics
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• 제29권1호
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• pp.98-103
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• 2021

The demand for natural substances with anti-melanogenic activity is increasing due to the recent interest in skin whitening. Intensive investigation on the culture broth of Streptomyces sp. SCO-736, a marine bacterium from the Antarctica coast, has led to the isolation of a new natural product named antaroide (1). The chemical structure was established through the interpretation of MS, UV, and NMR spectroscopic data. Antaroide is a nine-membered macrolide with lactone and lactam moieties. To investigate its applicability in skin whitening cosmetics, its anti-melanogenic activity in B16F10 murine melanoma cells was examined. As a result, antaroide displayed strong inhibitory activities against melanin synthesis and also attenuated the dendrite formation induced by the α-melanocyte stimulating hormone (α-MSH). Antaroide suppressed the mRNA expression of the melanogenic enzymes such as tyrosinase, TRP-1 and TRP-2. This suggests that it may serve as a transcriptional regulator of melanogenesis. Collectively, the discovery of this novel natural nine-membered macrolide and its anti-melanogenic activity could give new insights for the development of skin whitening agents.

• 동의생리병리학회지
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• 제21권5호
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• pp.1243-1249
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• 2007

The aim of this study was to investigate the effect of ethanol extract of Fagopyrum esculentum on the melanogenesis. To determine whether ethanol extract of Fagopyrum esculentum suppress melanin synthesis in cellular level, B16F10 melanoma cells were cultured in the presence of different concentrations of Fagopyrum esculentum ethanol extract. In the present study, we examined the effects of Fagopyrum esculentum ethanol extract on cell proliferation, melanin contents, tyrosinase activity, expression of melanogenic enzyme proteins including tyrosinase, tyrosinase-related protein 1 (TRP-1) and tyrosinase-related protein 2 (TRP-2). Cell proliferation was slightly increased by treatment with ethanol extract of Fagopyrum esculentum $(25-200 {\mu}g/m{\ell}).$ The ethanol extract of Fagopyrum esculentum effectively suppressed melanin contents at a dose of $100 {\mu}g/m{\ell}).$ It was observed that the color of cell pellets was totally whitened compared with the control. The ethanol extract of Fagopyrum esculentum inhibited tyrosinase activity, regulate melanin biosynthesis as the key enzyme in melanogenesis. Using western blot analysis, the ethanol extract of Fagopyrum esculentum dose-dependently decreased tyrosinase and TRP-1 protein levels, and tyrosinase and TRP-1 were detected in similar manner. ${\alpha}-MSH$ leads to a stimulation of melanin synthesis through increase of tyrosinase activity, melanin contents and cytoplasmic dendricity. In this study, ethanol extract of Fagopyrum esculentum down-regulated the ${\alpha}-MSH$-induced tyrosinase activity, melanin contents and cytoplasmic dendricity. Regarding protein levels of the melanogenic enzymes, the amounts of tyrosinase and TRP-1 was increased after incubation with a-MSH. The treatment of ethanol extract of Fagopyrum esculentum decreased the ${\alpha}-MSH$-induced expression levels of tyrosinase and TRP-1. These results suggest that the ethanol extract of Fagopyrum esculentum exerts its depigmenting effects through the suppression of tyrosinase, TRP-1 and cytoplasmic dendricity. And it may be a potent depigmetation agent in hyperpigmentation condition.