• Archives of Pharmacal Research
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• 제24권4호
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• pp.307-311
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• 2001

The activation of NF-$\kappa$B induced by kojic Acid, an inhibitor of tyrosinase for biosynthesis of melanin in melanocytes, was investigated in human transfectant HaCaT and SCC-13 cells. These two keratinocyte cell lines transfected with pNF-$\kappa$B-SEAP-NPT plasmid were used to determine the activation of NF-$\kappa$B. Transfectant cells release the secretory alkaline phosphatase (SEAP) as a transcription reporter in response to the NF-$\kappa$B activity and contain the neomycin phosphotransferase (NPT) gene for the dominant selective marker of geneticin resistance. NF-$\kappa$B activation was measured in the SEAP reporter gene assay using a fluorescence detection method. Kojic Acid showed the inhibition of cellular NF-$\kappa$B activity in both human keratinocyte transfectants. It could also downregulate the ultraviolet ray (UVR)-induced activation of NF-$\kappa$B expression in transfectant HaCaT cells. Moreover, the inhibitory activity of kojic Acid in transfectant HaCaT cells was found to be more potent than known antioxidants, e.g., vitamin C and N~acetyl-L-cysteine. These results indicate that kojic Acid is a potential inhibitor of NF-$\kappa$B activation in human keratinocytes, and suggest the hypothesis that NF-$\kappa$B activation may be involved in kojic Acid induced anti-melanogenic effect.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2019년도 추계학술대회
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• pp.90-90
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• 2019

Hibiscus syriacus exhibited promising potential as a new source of food and colorants containing various anthocyanins. However, the function of anthocyanins from H. syriacus has not been investigated. In the current study, we evaluated whether anthocyanins from the H. syriacus varieties Pulsae and Paektanshim (PS and PTS) inhibit melanin biogenesis. B16F10 cells and zebrafish larvae were exposed to PS and PTS in the presence or absence of ${\alpha}$-melanocyte-stimulating hormone (${\alpha}$-MSH), and melanin contents accompanied by its regulating genes and proteins were analyzed. PS and PTS moderately downregulated mushroom tyrosinase activity in vitro, but significantly decreased extracellular and intracellular melanin production in B16F10 cells, and inhibited ${\alpha}$-MSH-induced expression of microphthalmia-associated transcription factor (MITF) and tyrosinase. PS and PTS also attenuated pigmentation in ${\alpha}$-MSH-stimulated zebrafish larvae. Furthermore, PS and PTS activated the phosphorylation of extracellular signal-regulated kinase (ERK), whereas PD98059, a specific ERK inhibitor, completely reversed PS- and PTS-mediated anti-melanogenic activity in B16F10 cells and zebrafish larvae, which indicates that PS- and PTS-mediated anti-melanogenic activity is due to ERK activation. Moreover, chromatography data showed that PS and PTS possessed 17 identical anthocyanins as a negative regulator of ERK. These findings suggested that anthocyanins from PS and PTS inhibited melanogenesis in vitro and in vivo by activating the ERK signaling pathway.

• Bulletin of the Korean Chemical Society
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• 제30권2호
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• pp.475-478
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• 2009

• Journal of Applied Biological Chemistry
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• 제57권4호
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• pp.365-371
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• 2014

본 연구는 제주도 해변의 벌판에서 드물게 자라는 산유자(Xylosma congesta) 잎 에탄올 추출물의 화장품 기능성 소재로서의 활용 가능성을 알아보기 위하여 cell culture model 및 in vitro assay system을 이용하여 미백, 주름개선, 항염증 및 항산화 활성을 분석하였다. 그 결과 산유자 잎 에탄올 추출물은 B16F10 세포에 세포독성 없이 효과적으로 ${\alpha}$-MSH에 의해 유도된 melanin 합성을 억제함으로써 높은 미백 활성을 가지는 것이 관찰되었다. CCD-986SK 세포를 이용하여 산유자 잎 에탄올 추출물의 주름개선 활성 능력을 procollagen 합성시험을 통해 분석한 결과, 양성 대조군으로 사용한 TGF-${\beta}$ 만큼은 아니지만 농도별로 각각 120, 122%의 procollagen 합성을 증가시키는 것을 통하여 산유자 잎 에탄올 추출물의 주름개선 활성을 확인하였다. 또한 RAW 264.7 murine macrophage를 이용하여 NO 생성 억제능을 분석함으로써 산유자 잎 에탄올 추출물의 항염증 활성 정도를 측정한 결과, LPS에 의해 유도된 NO 생성이 산유자 잎 에탄올 추출물의 농도 의존적으로 감소하는 결과를 얻을 수 있었다. DPPH 법, ABTS 법, ORAC 법을 이용하여 항산화 활성을 분석한 결과, $50{\mu}g/mL$의 농도에서 DPPH 및 ABTS 라디칼 소거능이 각각 75.2, 99.1% 증가하는 것이 확인 되었다. ORAC activity assay kit를 이용하여 항산화 활성을 측정한 결과 산유자 잎 에탄올 추출물의 농도에 따라 높은 항산화 활성이 관찰 되었고, DPPH와ABTS 라디칼 소거능 결과와 유사하게 $50{\mu}g$/mL의 농도에서 가장 높은 항산화 활성($573.74{\pm}0.79{\mu}M$ TE/g)을 나타내었다. 이러한 결과를 통하여 산유자 잎 에탄올 추출물이 높은 미백과 주름개선, 항염증 및 항산화 활성을 나타내는 것을 확인 할 수 있었으며, 기능성 화장품 소재로서의 활용가능성을 제시 할 수 있었다.

• Biomolecules & Therapeutics
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• 제29권4호
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• pp.445-451
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• 2021

Salicylamide, a non-steroidal anti-inflammatory drug (NSAID), is used as an analgesic and antipyretic agent. We have previously shown that several NSAIDs have anti-melanogenic properties in B16F1 melanoma cells. In contrast, we have found that salicylamide enhances melanin contents in B16F1 melanoma cells; however, the underlying mechanism is not known. Therefore, we investigated the mechanism through which salicylamide stimulates melanogenesis. Interestingly, salicylamide enhanced diphenolase activity in a cell-free assay. Western blotting and real-time RT-PCR revealed that salicylamide increased tyrosinase expression via transcriptional activation of the Mitf gene. Together, our results indicate that salicylamide could be used as an anti-hypopigmentation agent for skin and/or hair.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2022년도 추계학술대회
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• pp.118-118
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• 2022

Carex pumila Thunb. is a plant native to East Asia, Australia, and New Zealand. However, its effect on skin melanogenesis has not been investigated. In the present study, we evaluated its anti-melanogenic properties using B16F10 melanoma cells and zebrafish larvae in the presence or absence of α-melanocyte stimulating hormone (α-MSH). In this study we revealed that concentrations below 50 µg/mL did not induce any cytotoxicity in B16F10 melanoma cells and cardiotoxicity in zebrafish larvae. However, 50 µg/mL treatment significantly inhibited α-MSH-induced extracellular (from 181.24% α 0.62% to 105.15% α 0.31%) and intracellular melanin contents (from 119.8% α 1.2% to 53.4% α 1.7%) as well as intracellular tyrosinase activity (from 143.9% α 4.2% to 103.7% α 1.4%) in B16F10 melanoma cells. At 25 µg/mL and 50 µg/mL concentrations, it could significantly inhibit α-MSH induced hyperpigmentation in zebrafish larvae (from 100% α 2.3% to 60.7% α 1.3% and 47.5% α 1.9% respectively). Additionally, the extract suppressed α-MSH-induced cAMP-CREB-MITF signaling pathway and consequently inhibited tyrosinase expression in B16F10 melanoma cells. In conclusion, our results indicate that this plant extract could suppress the cAMP-CREB-MITF axis which consequently inhibits tyrosinase mediated melanogenesis.

• Journal of Applied Biological Chemistry
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• 제65권2호
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• pp.93-99
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• 2022

Lysosome organelle extract (LOE) was derived from egg whites. The lysosome is an intracellular organelle that contains several hydrolysis enzymes. Previous studies have reported that LOE performs important functions, such as melanin de-colorization and anti-melanin production in B16F10 melanoma cells. However, its principal molecular and cellular mechanisms have not been elucidated till date. In non-cytotoxic conditions, LOE significantly inhibited α-MSH induced melanin synthesis of murine B16F10 cells. The anti-melanogenic activity of LOE was mediated by suppressing the mRNA expression of tyrosinase enzyme, tyrosinase related protein-1/2 (TRP-1/2), and microphthalmia-associated transcription factor (MITF) genes. By performing western blot analysis, we found that LOE significantly attenuated melanogenesis. In this case, LOE helped in increasing extracellular receptor kinase (ERK) phosphorylation in α-MSH induced B16F10 cells. Furthermore, MITF is found to be a key regulatory transcription factor in melanin synthesis; it was down-regulated by LOE through ERK phosphorylation. In this experiment, PD98059 (MEK inhibitor) was used to check whether LOE directly regulated the activity of ERK. Although LOE exerted inhibitory effect on melanin synthesis, we could not observe this effect in PD98059-treated α-MSH induced B16F10. These results strongly indicate that LOE is related to ERK activation and MITF degradation in anti-skin pigmentation. Hence, LOE should be utilized as a whitening agent of skin in the near future.

• 한국자원식물학회지
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• 제35권6호
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• pp.704-712
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• 2022

Research on whitening materials using natural alternatives is actively being conducted. The aim of this study was to investigate the in vitro inhibitory effects of Ranunculus chinensis Bunge (RCB) on melanogenesis and associated enzymes, such as tyrosinase, tyrosinase-related protein (TRP)-1, and TRP-2 in B16F10 murine melanoma cells. We found that RCB extract significantly attenuated melanin synthesis and reduced the activity of intracellular tyrosinase, a rate-limiting melanogenic enzyme. Western blot analysis showed that RCB extract decreased the protein expression of tyrosinase and TRP-1. In addition, it significantly decreased the expression of microphthalmia-associated transcription factor (MITF), a key regulator of melanogenesis. Extracellular signal-regulated kinase (ERK) activation has been reported to be involved in the inhibition of melanogenesis. Thus, we investigated whether the hypopigmentary effects of RCB extract were related to the activation of ERK. RCB extract induced ERK phosphorylation in a dose-dependent manner. Furthermore, it markedly inhibited body pigmentation in a zebrafish model. Our results suggest that RCB extract inhibits melanogenesis by activating ERK pathway-mediated suppression of MITF and its downstream target genes, including tyrosinase. Therefore, RCB extract can be used as a whitening agent in the development of functional cosmetics.

• Bulletin of the Korean Chemical Society
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• 제29권1호
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• pp.43-46
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• 2008

The aim of present study was to examine the inhibitory effects of hydroxamic acid derivatives on the melanogenesis. We found that hydroxamic acid moiety was important for anti-melanogenic activity. Compounds 1a and 1b strongly inhibited melanin synthesis via deactivation of tyrosinase. Hydroxamic acid has metal ion chelating ability which is similar to that kojic acid, however, anti-tyrosinase mechanism of compounds 1a and 1b was different from that of kojic acid. They showed noncompetitive inhibition kinetics

• 대한화장품학회지
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• 제38권3호
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• pp.247-254
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• 2012

남극 지의류인 Ramalina terebrata에서 분리 정제된 라말린(${\gamma}$-glutamyl-N'-(2-hydroxyphenyl)hydrazide)은 이전 연구에서 강력한 항산화능을 확인하였다. 본 연구에서는 라말린의 추가적인 효능을 확인하기 위하여, 비암세포 세포주인 멜란에이 세포를 이용하여 라말린의 멜라닌 합성에 대한 효과를 확인하였다. 라말린은 세포 독성이 없는 농도에서, 멜란에이 세포에서 멜라닌 합성을 농도 의존적으로 감소시켰으며, 이러한 효과는 널리 사용되고 있는 미백제인 알부틴보다 우수하였다. 라말린은 무세포 타이로시네이즈의 활성을 직접 저해했을 뿐만 아니라, 세포 내의 타이로시네이즈의 활성도 저해하는 효과를 보였다. 라말린의 이러한 멜라닌 합성 저해의 기전 연구를 위하여, 멜라닌 합성에 중요한 단백질인 타이로시네이즈, TRP-1, TRP-2의 mRNA와 단백질 발현을 조사한 결과, mRNA양에는 영향을 주지 않고, 단백질의 발현은 감소되는 것으로 나타났다. 또한, 0.2 % 라말린을 포함한 제형을 사람피부에 도포하였을 때, 3주 후에 피부 밝기가 개선됨을 확인하였다. 이상의 결과를 통해서, 라말린은 타이로시네이즈의 직접적인 저해뿐만 아니라 멜라닌 합성과 관련된 단백질의 발현을 저해함으로써 미백효과를 나타낸다고 할 수 있으며, 이러한 효과를 인체시험을 통해 확인하였다. 따라서, 라말린은 멜라닌 합성을 저해하는 미백 소재로 활용될 수 있을 것으로 판단된다.