• 생명과학회지
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• 제31권11호
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• pp.1019-1027
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• 2021

천연물에서 피부 미백 효능을 가진 화합물을 개발하기 위한 노력이 많이 이루어지고 있다. 꽃송이 버섯(Sparassis crispa)은 β-glucan의 함량이 건조중량의 40% 이상으로서 항암과 면역증강 효과가 있는 것으로 입증되어 약용으로도 인정받고 있다. 이 연구에서는 S. crispa β-glucan의 멜라닌 생합성 및 tyrosinase 활성억제 효능을 확인함으로써 피부 미백제로서의 활용 가능성을 평가하고자 하였다. 외부 자극제인 α-melanocyte stimulating hormone (α-MSH)와 S. crispa β-glucan (10, 100, 1,000 ㎍/ml)을 B16F1 melanoma 세포에 투여하여 멜라닌 생성량과 tyrosinase 활성도를 측정하였다. Tyrosinase, tyrosinase related protein-1 (TRP-1), TRP-2, microphthalmia-associated transcription factor (MITF)의 발현정도에 대해서는 western blot으로 분석하였다. S. crispa β-glucan을 10, 100, 1,000 ㎍/ml의 농도로 투여하였을 때 α-MSH 만 투여군에 비하여 멜라닌 생성이 각각 13.9%, 18.7%, 39.5% 감소하였다. S. crispa β-glucan을 10, 100, 1,000 ㎍/ml의 농도로 투여하였을 때 β-glucan을 투여하지않은 α-MSH 유도군에 비하여 tyrosinase 활성도는 각각 15.6%, 26.9%, 43.2% 억제되었다. 또한 tyrosinase와 TRP-1, TRP-2, MITF 단백 발현도 효과적으로 감소시켰다. 본 연구 결과에 따르면 S. crispa β-glucan은 MITF 발현을 억제함으로써 tyrosinase 발현을 감소시켜 멜라닌 생성을 억제시킨 것으로 판단된다. 따라서 S. crispa β-glucan은 미백제로서 유용하게 활용할 수 있을 것으로 생각한다.

• 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.

• 생명과학회지
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• 제23권11호
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• pp.1336-1341
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• 2013

아시아에서 한방약초로 널리 알려진 당귀 추출물의 미백활성을 알아보기 위하여 tyrosinase 저해활성을 측정한 결과 1,000 ${\mu}g/ml$의 농도에서 70% 이상의 활성을 나타내었다. 또한 당귀 추출물에 대한 멜라노마 세포(B16F10)의 세포생존율을 확인한 결과 500 ${\mu}g/ml$의 농도에서 99% 이상의 세포생존율을 확인할 수 있었다. 미백 관련 인자인 MITF, TRP-1, TRP-2 및 tyrosinase의 mRNA 발현량을 측정한 결과 50 ${\mu}g/ml$의 농도에서 각각 85.7%, 123.9%, 68.8%, 208%로 당귀 추출물을 처리하지 않은 군보다 감소하였음을 확인할 수 있었다. 이러한 연구 결과에 따라 당귀 추출물이 melanin 합성과 관련이 있는 유전자 발현의 억제효과가 있음을 확인할 수 있었으며, 미백 화장품 소재로서의 가능성을 확인하였다.

• International Journal of Stem Cells
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• 제16권2호
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• pp.135-144
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• 2023

Background and Objectives: Melanocyte (MC), derived from neural crest stem cell (NCSC), are involved in the production of melanin. The mechanism by which NCSC differentiates to MC remains unclear. N6-methyladenosine (m6A) modification was applied to discuss the potential mechanism. Methods and Results: NCSCs were isolated from hair follicles of rats, and were obtained for differentiation. Cell viability, tyrosinase secretion and activity, and transcription factors were combined to evaluated the MC differentiation. RT-qPCR was applied to determine mRNA levels, and western blot were used for protein expression detection. Total m6A level was measured using methylated RNA immunoprecipitation (MeRIP) assay, and RNA immunoprecipitation was used to access the protein binding relationship. In current work, NCSCs were successfully differentiated into MCs. Fat mass and obesity associated gene (FTO) was aberrant downregulated in MCs, and elevated FTO suppressed the differentiation progress of NCSCs into MCs. Furthermore, microphthalmia-associated transcription factor (Mitf), a key gene involved in MC synthesis, was enriched by FTO in a m6A modification manner and degraded by FTO. Meanwhile, the suppression functions of FTO in the differentiation of NCSCs into MCs were reversed by elevated Mitf. Conclusions: In short, FTO suppressed the differentiating ability of hair follicle-derived NCSCs into MCs by m6A modifying Mitf.

• Journal of Applied Biological Chemistry
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• 제64권4호
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• pp.323-331
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• 2021

Lincomycin is a lincosamide antibiotic isolated from the actinomycete Streptomyces lincolnensis. Moreover, it has been found to be effective against infections caused by Staphylococcus, Streptococcus, and Bacteroides fragillis. To identify the melanin-inducing properties of lincomycin, we used B16F10 melanoma cells in this study. The melanin content and intracellular tyrosinase activity in the cells were increased by lincomycin, without any cytotoxicity. Western blot analysis indicated that the protein expressions of tyrosinase, tyrosinase related protein 1 (TRP1) and TRP2 increased after lincomycin treatment. In addition, lincomycin enhanced the expression of master transcription regulator of melanogenesis, a microphthalmia-associated transcription factor (MITF). Lincomycin also increased the phosphorylation of p38 mitogen-activated protein kinase (MAPK) and decreased the AKT phosphorylation. Moreover, the activation of tyrosinase activity by lincomycin was inhibited by the treatment with SB203580, which is p38 inhibitor. Furthermore, we also found that lincomycin-induced tyrosinase expression was reduced by H-89, a specific protein kinase A (PKA) inhibitor. These results indicate that lincomycin stimulate melanogenesis via MITF activation via p38 MAPK, AKT, and PKA signal pathways. Thus, lincomycin can potentially be used for treatment of hypopigmentation disorders.

• Journal of Applied Biological Chemistry
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• 제63권1호
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• pp.89-93
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• 2020

In this study, the efficacy of melanoma cell B16F10 was investigated using the Korean native plant Oplismenus undulatifolius (OU). First, the cell viability of the extract was more than 90% when treated with 15 ㎍/mL of phenolics from OU. The results showed that melanin biosynthesis and cellular tyrosinase synthesis were inhibited by treatment with α-melanocyte-stimulating hormone-stimulated mouse melanoma cell B16F10 at a concentration of 15 ㎍/mL of phenolics for cell-line efficacy. The expression of tyrosinase, tyrosinase-related protein (TRP)-1, TRP-2, and microphthalmia transcription factor (MITF) protein was confirmed by western blot to investigate the effect of phenolics from OU on melanin biosynthesis. When treated with phenolics from OU 15 ㎍/mL, tyrosinase, TRP-1, TRP-2, and MITF decreased the protein expression level. In particular, tyrosinase, TRP-1, and MITF inhibited the production amount to a level similar to that of the non-treated normal group, indicating that the effect was excellent. Therefore, phenolics from OU acts as an inhibitor of tyrosinase, TRP-1, TRP-2, and its transcription factor MITF, and participates in melanin biosynthesis mechanism. These results suggested the potential for development as a material.

• The Korean Journal of Physiology and Pharmacology
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• 제26권2호
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• pp.113-123
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• 2022

Diarylpropionitrile (DPN), a selective agonist for estrogen receptor β (ERβ), has been reported to regulate various hormonal responses through activation of ERβ in tissues including the mammary gland and brain. However, the effect of DPN on melanogenesis independent of ERβ has not been studied. The aim of this study is to examine the possibility of anti-melanogenic effect of DPN and its underlying mechanism. Melanin contents and cellular tyrosinase activity assay indicated that DPN inhibited melanin biosynthesis in alpha-melanocyte stimulating hormone-stimulated B16F10 melanoma cell line. However, DPN had no direct influence on in vitro tyrosinase catalytic activity. On the other hand, 17β-estradiol had no effect on inhibition of melanogenesis, suggesting that the DPN-mediated suppression of melanin production was not related with estrogen signaling pathway. Immunoblotting analysis showed that DPN down-regulated the expression of microphthalmia-associated transcription factor (MITF), a central transcription factor of melanogenesis and its down-stream genes including tyrosinase, tyrosinase-related protein (TRP)-1, and TRP-2. Also, DPN attenuated the phosphorylation of protein kinase A (PKA) and cAMP-response element-binding protein (CREB). Additionally, DPN suppressed the melanin synthesis in UVB-irradiated HaCaT conditioned media culture system suggesting that DPN has potential as an anti-melanogenic activity in physiological conditions. Collectively, our data show that DPN inhibits melanogenesis via downregulation of PKA/CREB/MITF signaling pathway.

• The Korean Journal of Physiology and Pharmacology
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• 제16권4호
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• pp.287-291
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• 2012

This study investigated the effects of proline-serine (PS) and valine-serine (VS) dipeptides on melanogenesis in Mel-Ab cells. Proline-serine and VS significantly inhibited melanin synthesis in a concentration-dependent manner, though neither dipeptide directly inhibited tyrosinase activity in a cell-free system. Both PS and VS down-regulated the expression of microphthalmia-associated transcription factor (MITF) and tyrosinase. In a follow-up study also described here, the effects of these dipeptides on melanogenesis-related signal transduction were quantified. Specifically, PS and VS induced ERK phosphorylation, though they had no effect on phosphorylation of the cAMP response element binding protein (CREB). These data suggest that PS and VS inhibit melanogenesis through ERK phosphorylation and subsequent down-regulation of MITF and tyrosinase. Properties of these dipeptides are compatible with application as skin-whitening agents.

• 대한수의학회지
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• 제58권1호
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• pp.1-7
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• 2018

Genus Artemisia occurs as a hardy plant and has a wide range of culinary and medicinal features. In this study, we aimed to describe the melanin inhibitory activity of one Artemisia species, i.e., Artemisia capillaris Thunb. Ethanol extracts of fermented Artemisia capillaris (Art.EtOH.FT) and non-fermented Artemisia capillaris (Art.EtOH.CT) were tested for their ability to inhibit tyrosinase activity and melanin pigmentation. Both extracts showed dose-dependent inhibition against ${\alpha}$-melanocyte stimulating hormone-stimulated melanin formation and tyrosinase activity, without cytotoxicity. At $100{\mu}g/mL$, both extracts showed greater inhibition than kojic acid, the positive control. Protein expressions of microphthalmia-associated transcription factor (MITF), tyrosinase (TYR), tyrosinase-related protein 1 (TRP-1), and tyrosinase-related protein 2 (TRP-2) at the transcriptional level were determined by using real-time and semi-quantitative polymerase chain reaction. To complete the mechanistic study, presences of upstream elements of MITF, the phosphorylated-extracellular signal-regulated kinase (p-ERK), and phosphorylated-mitogen-activated protein kinase kinase (p-MEK) were confirmed by using western blot analysis. Expressions of p-TYR, p-TRP-1 and p-TRP-2, downstream factors for p-ERK and p-MITF, were translationally inhibited by both extracts. Art.EtOH.FT induced more potent effects than Art.EtOH.CT, especially signal transduction effects. In summary, Artemisia capillaris extracts appear to act as potent hypopigmentation agents.

• 한국가금학회지
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• 제41권1호
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• pp.29-37
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• 2014

닭을 구분하는데 있어 가장 눈에 띠는 것이 모색이며, 모색 관련 유전자인 MC1R, MITF, TYRP1의 SNP에 따른 유전자형을 확인하고, 각 품종별로 구별이 가능한 SNP 마커를 개발하고자 하는데, 본 연구의 목적이 있다. 마커들을 조합으로 haplotype을 보았을 때, MC1R 유전자의 SNP 조합에서 CGG type일 경우, 재래닭 만을 특별히 구별할 수 있었으며, TAG, TGG, TAA type일 경우에는 아라카나 만을 구별할 수 있었고, CAA type의 경우, 레그혼 만을 특이적으로 구별할 수 있었다. TYRP1 유전자의 SNP 조합에서는 TTTCA, CCTCA type의 경우, 레그혼 만을 구별할 수 있으며, CTTTA type의 경우, 오골계 만을 특이적으로 구별할 수 있었다. MC1R 유전자의 SNP 조합으로 재래닭, 레그혼, 아라카나를 구별할 수 있었고, TYRP1 유전자의 각각의 SNP 및 조합으로 4가지 품종 모두 구별할 수 있었다. 이렇게 품종 간의 유전적 다형성에 대한 연구가 더 많이 진행된다면, 단순 모색만으로 품종을 구별하기보다는 분자생물학적으로 품종 간의 차이를 이해할 수 있게 될 것이라고 생각된다.