• 대한본초학회지
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• 제34권2호
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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.

• KSBB Journal
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• 제25권5호
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• pp.478-482
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• 2010

죽여 추출물의 미백 효과를 확인하기 위하여 용매와 추출방법을 달리한 6가지의 추출물을 얻어 tyrosinase 활성 억제시험을 하였다. 그 결과 메탄올로 추출한 죽여 추출물에서 tyrosinase 활성 억제 효과가 가장 우수한 것을 확인 하였다. 세포 수준에서 죽여 추출물의 melanin 생성 억제 효과를 알아보기 위해서 B16 melanoma 세포를 이용하여 MTT assay를 실시하였고, 50 ppm의 농도 까지는 세포 독성이 나타나지 않은 것을 확인하였다. 죽여 추출물에 의한 세포 내 melanin 합성 저해 효과를 측정하기 위해 melanin contents assay를 하였고, 10 ppm 추출물을 처리한 extra cellular에서는 control 대비 68%, intra cellular에서는 74% 수준으로 melanin 합성이 저해됨을 확인하였다. 이차원전기영동을 이용하여 B16 melanoma 세포의 proteome을 분석 해 본 결과, 죽여 추출물 을 투여하지 않은 대조군에서는 171개, 추출물을 투여한 것에서는 282개의 spot을 확인하였고, 120개의 spot matching 을 확인하였다. 이 중 미백에 관련된 12개의 단백질 동향분석을 통하여, 죽여 추출물이 melanin 합성 전이나 합성 중에 관련된 메커니즘에 관련하는 것을 알 수 있었다.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2011년도 임시총회 및 추계학술발표회
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• pp.16-16
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• 2011

Melanogenesis is a well-known physiological response of human skin that may occur because of exposure to ultraviolet light, for genetic reasons, or due to other causes. In our effectors to find new skin lightening agents, acanthoic acid (AA) was investigated for its ability to inhibit melanogenesis. The effects of AA isolated from A.koreanumun the expression of $\alpha$-MSH-induced melanogenic factors (tyrosinase, tyrosinase related protein (TRP)-1, TRP-2 and MITF (microphthalmla-associated transcriptional factor)) were investigated in murine B16F10 melanoma cells. The results indicate that AA was an effective inhibitor of melanogenesis in B16F10 cells. To elucidate the mechanism of the effect of AA on melanogenesis, we performed Western blotting for melanogenic proteins. AA inhibited melanogenic factors (tyrosinase, TRP-1, TRP-2) expressions. In this study, we also confirmed that AA decreased the protein level of MITF proteins, which would lead to a decrease of tyrosinase and related genes in B16F10 melanoma cells. In order to apply AA to the human skin, the cytotoxic effects of the AA were determined by MTT assays using human keratinocyte HaCaT cells. Based on these results, we suggest that AA be considered possible anti-melanogenic agent and might be effective against hyperpigmentation disorders for the topical application.

• 원예과학기술지
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• 제31권6호
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• pp.813-820
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• 2013

울릉도 자생 삼나물(Aruncus dioicus)은 최근 monoterpenoids 생리활성 물질이 밝혀지면서 여러 가지 항산화와 관련된 생리활성이 확인되고 있다. 삼나물 ethyl acetate 분획물에 의한 미백효과를 검증하기 위하여 흑색세포종인 B16F10을 이용하여 실험을 진행하였다. 삼나물 ethyl acetate 분획물(ADE)의 세포독성을 측정하기 위하여 MTT assay를 실시한 결과, ADE $500{\mu}g{\cdot}mL^{-1}$ 이상의 농도에서 미비한 세포독성(10% 이상)을 확인하였으며, 이후 실험에서는 5, 10, $50{\mu}g{\cdot}mL^{-1}$ 농도가 세포 내에서 tyrosinase 활성과 멜라닌 양의 변화를 측정하였다. 그 결과 ADE 농도에 따라 tyrosinase 활성이 감소하고 총 멜라닌 함량이 감소하는 것을 확인하였다. 특히 $50{\mu}g{\cdot}mL^{-1}$에서 35.6% tyrosinase 활성억제, 58.8% 멜라닌 함량이 감소하는 것을 확인하였다. 또한 ADE에 의해 미백과 관련된 tyrosinase, tyrosinase related protein 1(TRP1), TRP2, microphthalmia associated transcription factor(MITF) 및 그 상위 단계인 cAMP와 protein kinase A(PKA)의 단백질 양이 감소하여 cAMP response binding protein(CREB)의 인산화는 감소하고 extracellular signal related kinase(ERK)의 인산화는 증가하는 것을 확인하였다. 본 연구에서는 울릉도 자생 삼나물의 미백효과에 관한 효능을 확인하고 기능성화장품의 소재로서의 활용 가능성이 있음을 확인하였다.

• 생약학회지
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• 제52권2호
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• pp.84-91
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• 2021

This study was carried out to investigate the melanin synthesis inhibitory effect of Aspergillus fumigatus fermented extract. In this study, we revealed the effects of A. fumigatus fermented extract on melanin contents, mushroom tyrosinase activity, and expression levels of mRNA and proteins of melanogenesis-related gene in B16F10 melanoma cells. A. fumigatus fermented extract inhibited both melanin contents and tyrosinase activity. In addition, the expression level of mRNA or proteins of melanogenesis was down-regulated in the A. fumigatus fermented extract treated B16F10 cells with dose-dependent manner. Moreover, when the clinical test was conducted, it was confirmed that the use of the fermented extract of A. fumigatus for 8 weeks improved skin brightness 1.586 times brighter and skin melanin 1.331 times better compared to the control product. Taken together, our results suggest that A. fumigatus fermented extract has melanogenesis inhibitory effect and whitening activity, thus it showed the possibility for using as a functional whitening cosmetic resource.

• 한국식품영양과학회지
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• 제39권10호
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• pp.1418-1424
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• 2010

본 연구에서는 정공피 물 추출물의 미백 효과를 확인하기 위하여 항산화 효능을 검증하였고, 멜라닌 생성의 첫 단계인 tyrosinase 활성, mouse melanoma B16BL6 세포 생존율 및 TRP-1, TRP-2 mRNA 발현 정도를 측정하였다. 정공피 물추출물은 DPPH 및 hydroxy 라디칼 소거 작용을 통하여 높은 항산화 효능을 나타내었고, tyrosinase, TRP-1, TRP-2의 활성을 효과적으로 억제하는 것을 확인하였다. 이는 정공피 물 추출물이 직접적으로 B16BL6 melanoma 세포의 tyrosinase 단백질의 활성을 저해시킴으로써 멜라닌 생성을 억제하는데 효과적임을 의미한다. 결론적으로 정공피 추출물은 항산화 활성과 미백 효과가 우수하여 기능성 화장품의 천연 소재로서 활용 가능성이 매우 높은 것으로 판단된다.

• 생명과학회지
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• 제27권5호
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• pp.536-544
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• 2017

미선나무 미성숙 종자는 물푸레나무과의 관목으로 전세계적으로 1속 1종의 중요한 식물자원이다. 대한민국에서는 미선나무 자생지를 보존하고 멸종위기식물로 보호하고 있다. 이러한 이유로 미선나무 미성숙 종자에 대한 연구는 미비하다. 본 연구에서 미선나무 미성숙 종자의 항산화 활성과 미백 관련 단백질인 tyrosinase, TRP-1, TRP-2, MITF의 단백질 발현 및 mRNA 수준의 발현 억제 활성을 확인하였다. 미선나무 미성숙 종자는 활성산소종에 효과가 뛰어났으며, 활성산소종은 노화, 염증, 암 등 다양한 질병을 야기시킨다. 항산화 활성은 DPPH, ABTS 라디칼 소거활성 및 환원력을 평가하였으며, 이러한 활성은 페놀류 화합물과 관계가 있는 것으로 알려져 있다. 페놀류 화합물은 천연 폴리페놀이라고 불리는 파이토케미칼로서 다양한 환경적 요인에 의한 식물 방어 기작의 일환으로 생성되는 2차 대사산물이다. 페놀류 화합물은 노화, 항암을 포함한 많은 인간의 건강에 긍정적인 영향을 준다고 알려져 있다. 미선나무 미성숙 종자는 tyrosinase, TRP-1, TRP-2 단백질 및 mRNA를 조절하였으며, 이러한 요인은 멜라닌 생합성에 중요한 역할을 한다. 또한 microphthalmia-associated transcription factor (MITF)의 단백질 및 mRNA를 억제하였다. MITF는 Tyrosinase, TRP-1, TRP-2의 발현과 전사에 연관된 인자로 알려져 있다. 미선나무 미성숙 종자의 미백활성, 페놀류 화합물, 항산화 활성 사이의 연관관계를 확인하였으며, 결론적으로 미선나무 미성숙 종자는 천연 식물 자원으로부터 얻을 수 있는 항산화제 및 피부 미백을 위한 기능성 화장품 원료로 사용될 수 있다.

• 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.

• Molecular & Cellular Toxicology
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• 제4권1호
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• pp.85-91
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• 2008

These days there is a constant possibility of exposure to UV radiation which can cause abnormal production of melanin and result in skin disease such as hyperpigmentation and melanoma. Many materials were investigated for skin whitening and protection against UV radiation. In this study, we assessed the melanogenesis inhibitory activities of Korean Red Ginseng (KRG, Ginseng Radix Rubra) and Ginkgo (EGb 761 Ginkgo Biloba) in an attempt to develop a new skin whitening agent derived from natural products. B16F10 melanoma cells were treated for 48 hr with KRG and EGb 761. The inhibitory effect on melanogenesis was measured and related cytokines and proteins expression were also investigated by RT-PCR and Western blotting. In addition, we also assessed the effects of these substances on the skin of C57BL/6 mice. Cell growth, melanin content and tyrosinase activity were inhibited effectively in B16F10 melanoma cells treated with KRG and EGb 761. Moreover, tyrosinase mRNA expression was inhibited clearly and melanogenesis related proteins (MRPs) containing tyrosinase, TRP1 and TRP2 were also reduced by KRG and EGb761, while cytokines such as IL-$1{\beta}$ and IL-6 were induced. In the case of UV irradiated mice, we observed induction of cytokine mRNA levels and reduction of MRPs mRNA expression. In addition, a decrease in pigmentation from treatment with KRG and EGb 761 on the skin of mice was observed. These results indicate that KRG and EGb 761 inhibit melanogenesis in B16F10 cells and have display protective activities against UVB. Therefore, we suggest that KRG and EGb 761 are good candidates to be used as whitening agents and UVB protectors for the skin.

• Journal of Applied Biological Chemistry
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• 제64권4호
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• pp.447-451
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• 2021

In this study, we discovered for the first time that linarin, a flavonoid compound, enhances melanin biosynthesis in B16F10 cells, and subsequently elucidated the underlying mechanism of linarin-induced melanogenesis. Linarin showed no cytotoxicity at a concentration of 42 μM and significantly increased intracellular tyrosinase activity and melanin content in B16F10 cells. Mechanistic analysis showed that linarin increased the expression of tyrosinase, tyrosinase-related protein 1 (TRP-1), and microphthalmia-associated transcription factor (MITF) that are related to melanogenesis. Moreover, linarin decreased the phosphorylation of extracellular signal-regulated kinase (ERK) and protein kinase B (AKT). Finally, we evaluated the effect of the structure-activity relationship of linarin and its aglycone on melanogenesis. The results indicated that linarin enhances the expression of melanogenic proteins by activating MITF expression via the modulation of mitogen-activated protein kinase (MAPK), phosphatidylinositol 3-kinase (PI3K), and protein kinase B signaling pathways in B16F10 cells, thereby enhancing melanogenesis.