• 생약학회지
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• 제52권2호
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• pp.99-104
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• 2021

This study was performed to elucidated the inhibitory effects of 6,8-diprenylorobol on melanin synthesis by measuring the levels of cell viability, mRNA expression, tyrosinase activity, and melanin production in the B16F10 cell line. The effects of 6,8-diprenylorobol on tyrosinase-related protein 1 (TYRP1), TYRP2, tyrosinase (TYR), and microphthalmia-associated transcription factor (MITF) mRNA expression levels and melanin content were determined. Quantitative real-time RT-PCR shows that 6,8-diprenylorobol decreases the mRNA expression levels of TYRP1, TYRP2, TYR, and MITF in B16F10 cell line, resulting in lower levels of melanin production compared to α-MSH-treated B16F10 cells. Tyrosinase activity assays reveal that 6,8-diprenylorobol decreases melanin production in B16F10 cells. These results demonstrate the whitening effects of 6,8-diprenylorobol on B16F10 cells; thus, 6,8-diprenylorobol is a potent ingredient for skin whitening. Further research is needed on the mechanism of action of 6,8-diprenylorobol. Such research will benefit not only cosmetics, but also the health food and medical industries.

• Archives of Pharmacal Research
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• 제22권3호
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• pp.232-236
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• 1999

In this study, we present evidence that cotreatment of aloesin and arbutin inhibits tyrosinase activity in a synergistic manner by acting through a different action mechanism. Aloesin or arbutin similarly inhibited enzyme activity of human- and mushroom-tyrosinases with an IC50 value of 0.1 or 0.04 mM, respectively. Lineweaver-Burk plots of the enzyme kinetics data showed that aloesin inhibited tyrosinase activity noncompetitively with a Ki value of 5.3 mM, whereas arbutin did it competitively (Maeda, 1996). We then examined whether cotreatment of these agents inhibits the tyrosinase activity in a synergistic manner. The results showed that 0.01 mM aloesin in the presence of 0.03 mM arbutin inhibited activity of mushroom by 80% of the control value and the reverse was also true. The inhibitory effects were calculated to be synergistic according to the B rgi method. Taken together, we suggest that aloesin along with arbutin inhibits in synergy melanin production by combined mechanisms of noncompetitive and competitive inhibitions of tyrosinase activity.

• 한방안이비인후피부과학회지
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• 제25권1호
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• pp.1-11
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• 2012

Objective : Paeoniae radix alba(PRA) can enrich the blood and regulate menstruation, astringe yin and arrest sweating, calm the liver and arrest pain. This study was designed to investigate effects of PRA on skin whitening and elasticity using melanoma cells. Methods : In this experiment, effect of PRA on cell viability, inhibition of melanin synthesis and inhibitory effect on tyrosinase and elastase. Results : 1. More than $1,000\;{\mu}g/ml$ of PRA treated group showed lowered proliferation rates significantly compared to non-treated control group. 2. All of treated groups were lower levels of melanin synthesis respectively. 3. PRA did not show inhibitory effect on tyrosinase activities in vitro. But, PRA suppressed tyrosinase activities in B16F10 cells significantly. 4. PRA suppressed elastse type 1 activities in dose-dependent manner in vitro. But, PRA slightly suppressed elastase type 4 activities in vitro, and PRA also slightly suppressed elastase activities in vivo. Conclusion : These results suggest that PRA can inhibit melanin synthesis through ihhibitory action on tyrosinase activity and inhibt elastase activity, and also suggest that these results can be used for the study on maintaining skin whitening or elasticity.

• 대한한방부인과학회지
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• 제22권4호
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• pp.1-18
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• 2009

Purpose: This study was performed to elucidate the inhibitory effect of Mibaeksan (MB) on melanin synthesis in B16F10 mouse melanoma cell. Methods: To demonstrate the inhibitory effects of MB on melanin synthesis, we measured the amount of released and produced melanin in B16F10 melanoma cell. Also, we evaluated tyrosinase-activity in vitro as well as in B16F10 melanoma cell. And to investigate the action mechanism, we assessed the gene expression of tyrosinase, TRP-1, TRP-2, MMP-2, PKA, $PKC{\beta}$, ERK-1 ERK-2, AKT-1 and MITF in B16F10 melanoma cells. Results: 1. MB decreased the release and production of melanin in B16F10 melanoma cells. 2. MB decreased tyrosinase activity in vitro and in B16F10 melanoma cells. 3. MB decreased the expression of tyrosinase, TRP-1, TRP-2, PKA, $PKC{\beta}$ and MMP-2 in B16F10 melanoma cells. 4. MB increased the expression of ERK-1, ERK-2 and AKT-1 in B16F10 melanoma cells. 5. MB decreased the expression of MITF in B16F10 melanoma cells. Conclusion: From these results, it may be concluded that MB has the antimelanogenetic effects.

• 한국약용작물학회지
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• 제15권6호
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• pp.434-436
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• 2007

멜라닌 생합성은 tyrosinase의 작용으로 인한 tyrosine의 산화로부터 시작되어 dopaquinone 및 dopachrome가 생성되고 dopachrome는 다시 dihydroxyindole과 dopachrome tautomerase에 의한 dihydroxyindole-2-carboxylic acid 로 전환되는 과정을 거쳐 진행된다 (Alaluf et al.; 2001). 도깨비 부채 뿌리 추출물의 멜라닌 생합성 저해 기전을 검정하기 위하여 melan-a 세포에서의 tyrosinase 및 TRP-2 발현량을 western immunoblotting하여 측정한 결과 도깨비 부채 뿌리 추출물은 TRP-2 발현량에는 영향을 미치지 않으나 tyrosinase의 발현량을 현저히 감소시킴을 확인 할 수 있었고 따라서 이전의 연구와 종합하여 볼 때 도깨비 부채 뿌리 추출물은 멜라닌 생합성의 초기에 주요한 역할을 하는 tyrosinase의 활성 및 발현을 모두 억제함으로써 멜라닌 생성을 감소시키는 것으로 사료된다. 또한 도깨비 부채 뿌리 추출물은 brown guinea pig 피부에서 3% 농도로 도포 시 발적이나 이상 징후를 일으키지 않고 자외선으로 유도된 색소 침착을 감소 시켜 피부 미백 용도의 기능성 원료로써 큰 가능성을 가지고 있는 것으로 판단된다.

• 한방안이비인후피부과학회지
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• 제30권2호
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• pp.38-50
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• 2017

Objectives : This study was designed to investigate effects of Lonicera Flos Extracts(LFE) on whitening using B16F10 cell lines. Methods : In this experiment, we observed effects of LFE on cell viability, inhibitory effects of melanin synthesis, inhibitory effect on tyrosinase, tyrosinase activity, superoxide dismutase(SOD)-like activity and mRNA expression. Results : In results, more than $2000{\mu}g/ml$ of LFE treated group showed lowered cell viability rates significantly compared to albutin treated group. More than $2000{\mu}g/ml$ of LFE treated groups were lower levels of melanin synthesis. Inhibitory effects of melanin production showed in $1000{\mu}g/ml$ of LFE treated group. $1,000{\mu}g/ml$ of LFE treated group significantly suppressed tyrosinase activities in vitro. LFE and albutin treated group significantly decreased tyrosinase activity compared to non treated group. SOD-like activity of LFE treated group was lower than vitamin C treated group but increased depending on concentration. $500{\mu}g/ml$ of LFE treated group and $1,000{\mu}g/ml$ of LFE treated group was significantly increased. Tyrosinase mRNA expression of ${\alpha}$-MSH and LFE $250{\mu}g/ml$ treated group significantly decreased compared to ${\alpha}$-MSH treated group. Conclusions : These results suggest that LFE can inhibit melanin synthesis through inhibitory action on tyrosinase activity. And LFE suppressed tyrosinase activities B16F10 cells significantly. So I suggest LFE can apply to whitening.

• 대한화장품학회지
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• 제27권1호
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• pp.111-118
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• 2001

A novel melanogenic inhibitory compound, Phytoclear-EL1 (5,10-diacetyl-3-benzoyllathyrol) was isolated from Euphorbiae lathyridis. The effects of EL1 on culture normal human melanocytes (NHM) were assessed. EL1 reduced the melanin synthesis of NHM by 40% at a concentration of 2 $\square$g/ml without any apparent cytotoxicity. We also have found that the treatment of the cells with EL1 decreased the tyrosinase activity by 29% in situ. To elucidate the action mechanism of EL1, we investigated the changes in mRNA levels of tyrosinase, TRP-1 and TRP-2 using RT-PCR technique. AS a result, the mRNA levels of tyrosinase, TRP-1 and TRP-2 were markedly reduced by EL1 treatment. These results suggest that Phytoclear-EL1 is a novel whitening agent that is effective in human melanocytes.

• Applied Microscopy
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• 제41권3호
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• pp.169-177
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• 2011

홍차 열수추출물의 멜라닌 합성 저해능과 작용기전을 알아보기 위해 in vitro 실험으로 항산화능을 확인한 후, melan-a 세포를 사용하여 멜라닌 합성 저해능, tyrosinase 활성 저해능 및 발현저해능 등에 대한 실험을 실시하여 다음과 같은 결과를 얻었다.(1) 항산화능 실험에서 홍차 열수추출물의 총 폴리페놀 및 플라보노이드 함량은 각각 102, 87 mg/g으로 양성대조군인 알부틴(25, 11 mg/g)보다 높았고, 전자공여능 역시 800 ${\mu}g$/mL에서 82%로 알부틴(48%)보다 높았다. (2) Melan-a 세포 실험에서 홍차 열수추출물은 수지상돌기의 발달과 멜라닌 생합성을 효과적으로 저해하였고, 멜라닌 합성 저해능과 tyrosinase 활성 저해능이 알부틴보다 유의하게 (p<0.001) 높았다. (3) Tyrosinase 단백질 발현 저해능 실험에서 알부틴은 발현에 유의한 영향을 미치지 않은 반면, 홍차 열수추출물은 발현을 유의하게 (p<0.001) 저해하였다. (4) 유전자 발현 저해능 실험에서 알부틴과 홍차 열수추출물 모두 tyrosinase, TRP-1과 TRP-2의 mRNA 발현에 영향을 미치지 않았다. 본 연구를 통하여, 홍차 열수추출물은 우수한 항산화능과 함께 멜라닌 합성 저해능을 확인하였으며, 이는 tyrosinase의 생합성을 저해시키기 보다는 tyrosinase 단백질 발현량을 줄이고 tyrosinase 활성을 감소시킴으로써 멜라닌 합성을 저해시키는 것으로 사료된다.

• 한국식품과학회지
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• 제44권1호
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• pp.76-81
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• 2012

본 연구에서는 3가지 홍화자, 향부자, 형개 추출물의 미백효과를 연구하였다. 멜라닌 생성 첫 단계인 tyrosinase 억제활성과 멜라닌 생성 억제효과를 측정한 결과, 홍화자 에탄올 추출물이 tyrosinase 활성과 B16F10 melanoma 세포의 멜라닌 생성 억제하였다. 그 결과, 홍화자 추출물은 B16F10 melanoma 세포에서 melanogenesis 따른 tyrosinase 형성 억제에 따른 멜라닌 합성 관련 인자 MITF, tyrosinase, TRP-1, TRP-2 의 발현을 억제함에 따라 홍화자 추출물의 미백효과를 확인하였다. 따라서 홍화자는 미백효과를 가진 천연 기능성 재료로서 가능성이 매우 높은 것으로 판단된다.

• 대한한방부인과학회지
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• 제22권3호
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• pp.66-82
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• 2009

Purpose: This study was performed to elucidate the inhibitory effect of Yukmijihwangtang-gagambang (YMG) on melanin synthesis in B16F10 mouse melanoma cell. Methods: To demonstrate the inhibitory effects of YMG on melanin synthesis, we measured the amount of released and produced melanin in B16F10 melanoma cell. Also, we evaluated tyrosinase-activity in vitro as well as in B16F10 melanoma cell. And to investigate the action mechanism we assessed the gene expressions of tyrosinase, TRP-1, TRP-2, MMP-2, PKA, PKC${\beta}$, ERK-1 ERK-2, AKT-1 and MITF in B16F10 melanoma cells. Results: 1. YMG decreased the release and production of melanin in B16F10 melanoma cells. 2. YMG decreased tyrosinase activity in vitro and in B16F10 melanoma cells. 3. YMG decreased the expression of tyrosinase, TRP-1, TRP-2, PKA, PKC${\beta}$ and MMP-2 in B16F10 melanoma cells. 4. YMG increased the expression of ERK-1, ERK-2, and AKT-1 in B16F10 melanoma cells. 5. YMG decreased the expression of MITF in B16F10 melanoma cells. Conclusion: From these results, we suggest that YMG inhibit melanin synthesis via tyrosinase inhibition and regulation of the gene expression in B16F10 melanoma cells.