• 한국식품과학회지
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• 제51권1호
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• pp.58-63
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• 2019

본 연구에서 자소엽 조다당 추출물의 미백 활성을 평가하기 위하여 tyrosinase 억제활성과 melanocyte 세포에서 멜라닌 함량 및 멜라닌 관련 단백질 발현 억제능을 수행하였다. 자소엽 조다당 추출물은 tyrosinase 억제능을 관찰한 결과 자소엽 조다당 추출물의 농도가 증가할수록 활성이 유의적으로 증가하는 것으로 나타났다. 이러한 미백 효과가 melanoma 세포인 B16F10 세포 내에서도 일어나는지 확인하기 위하여 세포내 멜라닌 함량 및 tyrosinase 억제능을 살펴보았다. 자소엽 조다당 추출물은 B16F10 세포에 대하여 세포독성이 없는 농도에서 진행 하였으며, 세포내 멜라닌 함량 및 tyrosinase 활성 저해를 보여주었다. 이는 자소엽 조다당 추출물이 세포 내 MITF 발현을 억제시킴으로서 tyrosinase, TRP-1 및 TRP-2 단백질 발현을 억제함에 따라 미백효과를 확인하였다. 이러한 결과로 자소엽 조다당 추출물의 미백 활성을 확인 할 수 있었으며, 미백 산업 분야 및 식품의 기능성 소재로 이용한다면 천연 미백 소재로써 활용될 수 있을 것으로 판단된다.

• Toxicological Research
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• 제33권1호
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• pp.55-62
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• 2017

We evaluated the antioxidant activity and anti-melanogenic effects of Oenothera laciniata methanol extract (OLME) in vitro by using melan-a cells. The total polyphenol and flavonoid content of OLME was 66.3 and 19.0 mg/g, respectively. The electron-donating ability, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical-scavenging activity, and superoxide dismutase (SOD)-like activity of OLME ($500{\mu}g/mL$) were 94.5%, 95.6%, and 63.6%, respectively. OLME and arbutin treatment at $50{\mu}g/mL$ significantly decreased melanin content by 35.5% and 14.2%, respectively, compared to control (p < 0.05). OLME and arbutin treatment at $50{\mu}g/mL$ significantly inhibited intra-cellular tyrosinase activity by 22.6% and 12.6%, respectively, compared to control (p < 0.05). OLME ($50{\mu}g/mL$) significantly decreased tyrosinase, tyrosinase-related protein-1 (TRP-1), TRP-2, and microphthalmia-associated transcription factor-M (MITF-M) mRNA expression by 57.1%, 67.3%, 99.0%, and 77.0%, respectively, compared to control (p < 0.05). Arbutin ($50{\mu}g/mL$) significantly decreased tyrosinase, TRP-1, and TRP-2 mRNA expression by 24.2%, 42.9%, and 48.5%, respectively, compared to control (p < 0.05). However, arbutin ($50{\mu}g/mL$) did not affect MITF-M mRNA expression. Taken together, OLME showed a good antioxidant activity and anti-melanogenic effect in melan-a cells that was superior to that of arbutin, a well-known skin-whitening agent. The potential mechanism underlying the anti-melanogenic effect of OLME was inhibition of tyrosinase activity and down-regulation of tyrosinase, TRP-1, TRP-2, and MITF-M mRNA expression.

• The Korean Journal of Physiology and Pharmacology
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• 제21권3호
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• pp.287-292
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• 2017

Vitiligo is an intriguing depigmentary disorder and is notoriously difficult to be treated. The ultimate goal of vitiligo treatment is to replenish the lost melanocytes by immigration from hair follicle and to restore the normal function of melanogenesis by residual melanocytes. There are two types of topical calcineurin inhibitors called tacrolimus and pimecrolimus, and are recommended as the first-line treatments in vitiligo. Although pimecrolimus is efficacious for the repigmentation of vitiligo, its intrinsic mechanisms have never been investigated in vitro. This research aimed to study the ability of pimecrolimus on stimulating melanogenesis, melanocyte migration and MITF (microphthalmia associated transcription factor) protein expression. Results showed that pimecrolimus at the dosages of 1, 10, $10^2$nM were neither mitogenic nor cytotoxic to melanocytes. The addition of pimecrolimus at 10, $10^2$ and $10^3nM$ significantly increased intracellular tyrosinase activity, which was consistent with the elevated content of melanin content at the same concentrations. The peak effect was seen at 72 h in response to $10^2$nM pimecrolimus. Results of the wound scratch assay and Transwell assays indicate that pimecrolimus is effective in facilitating melanocyte migration on a collagen IV-coated surface. In addition, MITF protein yield reached the highest by pimecrolimus at $10^2nM$. In brief, pimecrolimus enhances melanin synthesis as well as promotes migration of melanocytes directly, possibly via their effects on MITF protein expression.

• KSBB Journal
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• 제28권2호
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• pp.137-145
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• 2013

In order to develop new skin whitening agents, we prepared the $CH_2Cl_2$ layer (NGC) and BuOH layer (NGB) of 75% EtOH extract of the Nelumbinis nucifera Gaertner. We measured their tyrosinase inhibitory activity in vitro and melanin synthesis inhibitory activity in B16-F1 melanoma cells. They did not show inhibitory activity against mushroom tyrosinase but showed melanin synthesis inhibitory activity in a dose-dependent manner. In a melanin synthesis inhibition assay, NGC and NGB suppressed melanin production up to 52% and 46% at a concentration of $100{\mu}g/mL$, respectively. To elucidate the mechanism of the inhibitory effects of NGC and NGB on melanogenesis, we measured the expression of melanogenesis-related proteins by western blot assay. As a result, NGC suppressed the expression of tyrosinase, tyrosinase related protein 1 (TRP-1), tyrosinase related protein 2 (TRP-2), phosphorylated cAMP responsive element binding (p-CREB) protein, and microphthalmia associated transcription factor (MITF). And NGB inhibited the protein expression of tyrosinase and MITF, but had no significant effect on TRP-1, TRP-2, and p-CREB expression. Moreover, NGB increased the expression of phosphorylated extracellular signal-regulated kinase (p-ERK). In addition, we examined the inhibitory effect on the glycosylation of tyrosinase. As a result, NGC and NGB inhibited the activity of ${\alpha}$-glucosidase in vitro and the glycosylation of tyrosinase in B16-F1 melanoma cells. From these results, we concluded that NGC and NGB could be used as active ingredients for skin whitening.

• 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.

• 한국식품위생안전성학회지
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• 제30권3호
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• pp.258-264
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• 2015

본 연구는 윈터체리 추출물의 미백 활성을 검증하여 기능성 미백 소재로서의 가능성을 확인하였다. 먼저, DPPH, ABTS, FRAP를 이용한 항산화 활성 검증에서 윈터체리 추출물은 매우 높은 항산화 효과를 보였으며, 티로시나아 제의 활성에도 농도 의존적인 억제 효과를 보여주었다. 마우스 유래 B16-F10 멜라노마 세포를 이용한 멜라닌 생성에서 윈터체리 추출물이 미치는 영향을 확인한 결과, 세포 내에서 생합성 되는 멜라닌의 양이 상당히 감소하고 있음을 확인하였다. 또한, 멜라닌 생성을 더욱 유도하는 ${\alpha}-MSH$를 세포에 처리하였을 때, 윈터체리 추출물은 농도 의존적인 억제 효능을 보여주었다. 이러한 윈터체리 추출물의 멜라닌 생성 억제는 MITF 전사인자의 발현을 억제함으로써 일어나고, 결과적으로 멜라닌 생합성과 관련된 티로시나아제와 Tyrp-1 등의 단백질 발현을 감소시킴으로써 일어나게 됨을 확인하였다. 이러한 결과들로 볼 때, 윈터체리 추출물은 기존의 미백 원료들을 대체할 수 있을 뿐만 아니라 함께 사용할 경우 상승 효과를 낼 수 있는 새로운 미백 소재로 활용될 수 있을 것이다.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2021년도 춘계학술대회
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• pp.54-54
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• 2021

The Melanoma Research Coalition reported melanoma affects humans of various races. This study was conducted to confirm the inhibitory effect of melanogenesis in B16 F10 cells of Nypa fruticans Wurmb of ethyl acetate fraction (NEF). Nypa fruticans Wurmb is an important component of the East Asian mangrove vegetation. It belongs to Araceae family. Traditionally, N. fruticans was used to treat various diseases such as asthma, sore throat, liver disease, a pain reliever, and can also be used as sedative and carminative. The present study, the inhibitory effect on melanogenesis was determined by Western blotting and RT-qPCR. The level of expression of tyrosinase, TRP-1, and TRP-2 is regulated by microphthalmia-associated transcription factor (MITF) and cAMP, and cAMP affects the activity of protein kinase A (PKA). Activated PKA stimulates the phosphorylation of cAMP-reactive element-binding protein (CREB) in the nucleus, thereby increasing the amount of MITF expression and enhancing melanogenesis. Western blotting and RT-qPCR analysis showed that NEF treatment decreased the expression of tyrosinase. Similarly, TRP-1 and TRP-2 levels were decreased, which were decreased significantly at compared with the untreated control. Also, NEF attenuated the IBMX mediated increase in the intracellular cAMP level and the phosphorylation of PKA. In conclusion, NEF significantly inhibited the expressions of melanogenesis through cAMP/PKA/CREB signaling pathways.

• 공업화학
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• 제31권6호
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• pp.653-659
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• 2020

본 연구에서는 메톡시폴리에틸렌글리콜(methoxy polyethylene glycol)과 올레아놀산(Oleanolic acid) 유도체(mPEG-OA derivative)를 합성하였으며, 합성된 유도체에 대하여 수용액에서의 용해도와 멜라닌 생성억제 효과를 평가하였다. mPEG-OA 유도체의 합성된 구조는 1H NMR, 13C NMR 및 FT-IR로 확인하였다. 수용액에서 mPEG-OA 유도체와 OA의 용해도를 측정한 결과, mPEG-OA 유도체는 13 mg/mL, OA는 0.013 mg/mL로서, mPEG-OA 유도체의 수용성이 OA보다 1,000배 높게 나타냈다. 세포생존율은 B16F10 melanoma cells에서, mPEG-OA 유도체의 세포생존율(250 μM)이 OA로 처리한 세포생존율(62.5 μM)과 비교하여 4배 증가하였다. 멜라닌 생합성 억제 효과는 세포생존율이 영향을 받지 않는 농도에서 측정하였으며, mPEG-OA 유도체는 50 μM의 농도에서 36%, OA는 10 μM의 농도에서 35%의 억제 효과를 나타내었다. B16F10 melanoma cells에서 MITF (microphthalmia-associated transcription factor)의 발현 억제 수준은 mPEG-OA 유도체는 50 μM의 농도에서 59%, OA는 10 uM의 농도에서 49%의 억제 효과를 나타내었다. 종합적으로 mPEG-OA 유도체와 OA의 수용성 및 미백활성을 비교한 결과, mPEG-OA 유도체는 OA보다 뛰어난 수용성을 가지며, 멜라닌 생합성을 억제하는 효과를 나타냄으로써 미백 기능성 화장품 소재로서 응용 가능성이 있음을 시사한다.

• Journal of Nutrition and Health
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• 제52권2호
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• pp.168-175
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• 2019

Purpose: Barely sprout is a well-known oriental herbal medicine with a wide range of health benefits. Recent studies have provided scientific evidence of its therapeutic effects with expanded application. This study investigated anti-melanogenic effect of barley sprout water extract (BSE) in murine melanocyte B16F10. Methods: Various concentrations (0, 50, 125, and $250{\mu}g/mL$) of BSE and arbutin (150 ppm) were applied to B16F10 stimulated with or without alpha-melanocyte stimulating hormone (100 nM) for 72 hours. The whitening potency of BSE was determined altered cellular melanin contents. Activity and expression of tyrosinase and microphthalmia-associated transcription factor (MITF) were also assayed. Results: Experimental results revealed that treatment with BSE reduced cellular melanin production by approximately 40% compared to the control. Molecular findings supported that suppressed activity and expression of tyrosinase and MITF proteins by BSE were associated with declined cellular melanogenesis. Furthermore, anti-melanogenic effect of BSE ($250{\mu}g/mL$) was similar to that of arbutin, a commonly used whitening agent. Lastly, polyphenols including p-coumaric, ferulic, and vanillic acids were identified in BSE using HPLC analyses. They might be potential active ingredients showing such melanogenesis-reducing effect. Conclusion: BSE was evident to possess favorable anti-melanogenic potency in an in vitro model. As a natural food sourced material, BSE could be an effective depigmentation agent with potential application in pharmaceutical and cosmetic industries.

• 한국자원식물학회지
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• 제31권6호
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• pp.667-674
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• 2018

In this study, we evaluated the whitening activity of prethanol A and water extracts from Abeliophyllum distichum Nakai. The extracts were prepared using 0, 50, 70, and 100% prethanol A at $121^{\circ}C$, 1.2 atm for 15 minutes. To confirm effective extraction, the acteoside content of each extract was analyzed with the HPLC-PDA method. The antioxidant activity was evaluated using DPPH and ABTS scavenging activity assays, and the whitening activity was evaluated based on inhibitory activities on the protein and mRNA expression of tyrosinase, tyrosinase-related protein 1 (TRP-1), tyrosinase-related protein 2 (TRP-2), and microphthalmia-associated transcription factor (MITF) in B16 F10 cells. Each extract showed strong antioxidant and whitening activity. $IC_{50}$ values of antioxidant activity from each extract were in order of 100%, 70%, 50%, and 0%. In addition, whitening activity inhibited the protein and mRNA expression of melanin synthesis factor, following the same pattern as antioxidant activity. In conclusion, water and prethanol A extracts of A. distichum showed effective antioxidant and whitening activity and are thus considered to be valuable materials for whitening cosmetics. The results of this study will also provide basic data for the safe and efficient production of A. distichum as a cosmetic material.