• 약학회지
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• 제58권1호
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• pp.21-27
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• 2014

We have developed 8 peptide derivatives as potential MC1R antagonists and their inhibitory effects on ${\alpha}$-MSH induced cell growth in cultured normal human melanocytes (NHM) were investigated. From these experiments, the two most potent peptide derivatives, 5-phenylvaleric acid-(D)His-Arg-Trp-$(Lys)_6NH_2$ (P 6) and 5-phenylvaleric acid-(D)His-Arg-Trp-$(Lys)_9NH_2$ (P 7) were selected for further studies. In ${\alpha}$-MSH depleted NHM cells, we have found that the treatment with 1 ${\mu}M$ of these two peptide derivatives, P 6 and P 7, inhibited the cell proliferation induced by the addition of 1 nM ${\alpha}$- MSH by 70% and 72%, respectively. In NHM cells without previous ${\alpha}$-MSH depletion, 1 ${\mu}M$ treatment in the presence of 10 nM ${\alpha}$-MSH resulted in 70% (P 6) and 80% (P 7) decrease in cell growth and 64% (P 6) and 71% (P 7) reduction in melanin synthesis, respectively. The peptide derivatives P 6 and P 7 were proved to have no apparent cytotoxicity and inhibited the elevation of intracellular cAMP concentration triggered by ${\alpha}$-MSH. In conclusion, our data suggest that the peptide derivatives reported in this study, 5-phenylvaleric acid-(D)His-Arg-Trp-$(Lys)_6NH_2$ (P 6) and 5-phenylvaleric acid-(D)His- Arg-Trp-$(Lys)_9NH_2$ (P 7) strongly antagonize ${\alpha}$-MSH, inhibit cell proliferation and melanin synthesis, and lower the intracellular cAMP concentration, hence have a promising potential as a novel skin lightening agent.

• 한국환경성돌연변이발암원학회지
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• 제9권2호
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• pp.97-109
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• 1989

살균제 Captan, 살충제 Diazinon, MEP, Malathion 및 EPN 의 단독 처리 그리고 Captan 반복처리와 이들 살충제와 서로 다른 조합으로의 동시적, 시간적 복합처리가 E. coli WP2S(uvrA-, trp-) 균주의 돌연변이유발 빈도 및 생잔율에 미치는 영향을 검토하여 다음과 같은 결과를 얻었다. 1. Captan, Diazinon, MEP, Malathion, EPN 및 UV 의 동시적 복합처리는 이들 농약의 단독 처리에서 보다 E. coli WP2S (uvrA-, trp-)균주의 돌연변이유발빈도가 높았고 이들 농약의 단독처리효과를 합한 가산적 영향으로 나타났으며, 동일조건하에서 이들 농약의 처리농도를 배로 늘렸을 때는 배가효과를 보였으나 이 균주의 증가된 돌연변이 유발빈도는 cinnamaldehyde 30ng 및 60ng/ml 그리고 tannic acid 50ng/ml 배지로 각각 첨가했을 때 감소되었고 생잔율은 증가되었다.

• 한방안이비인후피부과학회지
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• 제20권3호
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• pp.1-13
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• 2007

Objective : This study was performed to assess the whitening effect of Bombysis Corpus on melanin synthesis. Methods : The whitening effects of Bombysis Corpus were examined by in vitro melanin production assay. We assessed inhibitory effects of Bombysis Corpus on melanin-release from B16F10, on melanin production in B16F10, on mushroom tyrosinase activity in vitro, on tyrosinase activity in B16F10, effect of Bombysis Corpus on the expression tyrosinase, TRP-1, PKA, ERK-1 ERK-2, AKT-1, MITF in B16F10. Results : 1. Bombysis Corpus inhibited melanin-release, melanin production in B16F10. 2. Bombysis Corpus inhibited tyrosinase activity in vitro and in B16F10. 3. Bombysis Corpus suppressed the expression of tyrosinase, TRP-1 in B16F10. 4. Bombysis Corpus suppressed the expression of PKA in B16F10. 5. Bombysis Corpus suppressed the expression of ERK-1, ERK-2, AKT-1 in B16F10. 6. Bombysis Corpus suppressed the expression of MITF in B16F10. Conclusion : The study shows that Bombysis Corpus inhibited melanin production on the melanogenesis.

• Biomolecules & Therapeutics
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• 제30권2호
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• pp.203-211
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• 2022

Melanogenesis is the production of melanin from tyrosine by a series of enzyme-catalyzed reactions, in which tyrosinase and DOPA oxidase play key roles. The melanin content in the skin determines skin pigmentation. Abnormalities in skin pigmentation lead to various skin pigmentation disorders. Recent research has shown that the expression of EMP2 is much lower in melanoma than in normal melanocytes, but its role in melanogenesis has not yet been elucidated. Therefore, we investigated the role of EMP2 in the melanogenesis of MNT1 human melanoma cells. We examined TRP-1, TRP-2, and TYR expression levels during melanogenesis in MNT1 melanoma cells by gene silencing of EMP2. Western blot and RT-PCR results confirmed that the expression levels of TYR and TRP-2 were decreased when EMP2 expression was knocked down by EMP2 siRNA in MNT1 cells, and these changes were reversed when EMP2 was overexpressed. We verified the EMP2 gene was knocked out of the cell line (EMP2 CRISPR/Cas9) by using a CRISPR/Cas9 system and found that the expression levels of TRP-2 and TYR were significantly lower in the EMP2 CRISPR/Cas9 cell lines. Loss of EMP2 also reduced migration and invasion of MNT1 melanoma cells. In addition, the melanosome transfer from the melanocytes to keratinocytes in the EMP2 KO cells cocultured with keratinocytes was reduced compared to the cells in the control coculture group. In conclusion, these results suggest that EMP2 is involved in melanogenesis via the regulation of TRP-2 expression.

• 한국응용과학기술학회지
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• 제38권5호
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• pp.1229-1240
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• 2021

은행나무 종자인 백과를 70% 에탄올로 추출하여 미백 효능을 규명하고 화장품 소재로서의 응용 가능성을 확인하고자 하였다. 백과 에탄올추출물 (GBE)을 B16F10 melanoma cells에 처치하여 멜라닌 생성과 tyrosinase 활성을 확인한 결과 유의한 수준의 멜라닌 생성 저해가 관찰되었고, 멜라닌 생성과정에 관여하는 주요 효소인 tyrosinase의 활성이 농도 의존적으로 억제됨이 관찰되었다. 멜라닌 생성 관련 주요 인자들의 단백질 발현과 mRNA 수준을 관찰한 결과, tyrosinase, tyrosinase-related protein-1, -2 (TRP-1, -2), microphthalmia-associated transcription factor (MITF)의 단백질 발현과 유전자 수준이 GBE의 처리에 의해 유의한 수준으로 저해되었다. 이 결과를 통해, 본 연구의 백과 에탄올추출물은 멜라닌 세포의 멜라닌 생성 관련 핵심 전사인자인 MITF의 조절을 통해 멜라닌 생성 억제 효과를 나타내는 것으로 보인다. 이에 따라 백과 에탄올추출물을 화장품 미백 기능성 소재로 활용 가능성이 있을 것으로 기대된다.

• 대한본초학회지
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• 제32권3호
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• pp.71-78
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• 2017

Objectives : The various grape extracts derived from grape pulp, seed and skin, containing various types of polyphenols and flavonoids, have been known to have anti-inflammatory, antioxidant and improve cardiovascular condition as well as sun's damaging effects. However, there have been rare reports of various beneficial effects of grape fruit stem extract (GFSE), one of the waste products of grapes. We investigated anti-inflammatory and melanogenesis inhibitory effects of GFSE. Methods : One-hundred gram of grape fruit stem was extracted with 80% ethanol at room temperature for 3 days. After filtration, the ethanol was removed using vacuum evaporator, then lyophilized to obtain the dry extract which was stored at $-20^{\circ}C$ until used. NO levels were measured by using Greiss reagent. Prostaglandin $E_2$ ($PGE_2$) production was measured by ELISA assay. The expression levels of iNOS, COX-2, TRP-1 and TRP-2 were evaluated by western blot analysis. Results : GFSE reduced the level of nitric oxide and prostaglandin $E_2$ ($PGE_2$) production in a dose-dependent manner, compared to control. Expressions of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) protein were also effectively inhibited by the GFSE. In a tyrosinase inhibitory activity, GFSE significantly reduced the tyrosinase activity and melanin content in a dose dependent manner, compared to control. GFSE also decreased the expression of tyrosinase related protein-1 (TRP-1) and tyrosinase related protein-2 (TRP-2), known as a melanocyte-specific gene product involved in melanin synthesis. Conclusions : Therefore, these results indicated that GFSE had powerful anti-inflammatory and anti-melanogenic effects.

• 대한화장품학회지
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• 제31권4호
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• pp.305-310
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• 2005

본 연구에서는 한방 천연 미백 소제를 개발하기 위하여 삼릉추출물의 미백 효과에 대하여 연구하였다. 삼릉추출물은 프리 라디칼 소거활성(DPPH)에서 $IC_{50}=638{\mu}g/mL$, superoxide radicals 소거활성은 $IC_{50}=21.7{\mu}g/mL$을 나타냈다. 삼릉추출물의 B16 melanoma 세포에서의 멜라닌 생합성 저해효과는 $100{\mu}g/mL 삼릉추출물을 48 h 처리한 세포에서는 멜라닌 합성량이 27% 감소되었다. 삼릉 추출물은 농도에 비례하여 B16 melanoma 세포내의 타이로시네이즈 활성을 저해하였으며, western blot을 이용하여 타이로시네이즈와 타이로시나아제 관련 단백질(TRP-1)의 발현감소를 확인하였다. 또한, RT-PCR을 이용하여 삼릉추출물이 멜라닌 생성 과정에 관련하는 유전자 발현을 조사한 결과 타이로시네이즈와 TRP-1의 mRNA발현을 억제하는 것을 확인하였다. 그러므로 삼릉추풀물은 항산화 효과와 함께 멜라닌 저해 효과가 우수하여 새로운 미백 화장품으로 응용할 수 있을 것으로 생각된다.

• Bulletin of the Korean Chemical Society
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• 제7권1호
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• pp.78-81
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• 1986

The photosensitized lysis of egg lecithin lipid membranes (liposomes) have been performed to UV-B light (270-320 nm) by L-tryptophan(L-Trp) and its peptide such as N-acetylphenylalanyl-L-tryptophan(NAPT) incorporated in the liposomes(ca. 0.1% by weight) or in the external buffer (0.1-0.3 mM). Requirement of oxygenation suggests that the lysis of liposomes is caused by the photosensitized oxidation of lipids. There was significant protection against lysis photosensitized by Trp in the external buffer by low concentration of ferricyanide (0.8 mM), but there was no effect on the lytic efficiency by $N_3^-$ which is singlet oxygen($^1O_2$) quencher, indicative of an electron transfer mechanism involved in the photosensitization. The small change of the lytic efficiency with increasing pH from 4 to 9 was interpreted by large target theory and subsequently indicates that superoxide($O_2^-$) may be an active intermediate for the oxidation. The efficiency of photosensitization of Trp was higher than that of NAPT under the same experimental condition. The weak lytic efficiency of liposomes photosensitized by NAPT was enhanced by incorporating NAPT in liposomes, but it was again quenched by ${\beta}$-carotene incorporated in the bilayer of liposomes. These results indicate that a portion of liposome lysis may be due to $^1O_2$ formation from the excited NAPT.

• Asian Pacific Journal of Cancer Prevention
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• 제13권9호
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• pp.4423-4426
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• 2012

Aim: XRCC1 and XPD are two major repair genes involved in nucleotide excision repair (NER), which is reported to be associated with risk of several cancers. We explored the association of XRCC1 and XPD polymorphisms with the risk of HCC. Methods: A total of 410 cases with HCC and 410 health controls were collected. XRCC1 Arg194Trp, XRCC1 Arg399Gln, XPD Lys751Gln and XPD Asp312Asn genotyping was performed by duplex polymerase-chain-reaction with the confronting-two-pair primer (PCR-CTPP) method. Results: XRCC1 194Trp/Trp was strongly significantly associated with an increased risk of HCC cancer when compared with the wide-type genotype (OR=2.26, 95% CI=(1.23-5.38). Individuals carrying the XRCC1 399Gln/Gln showed increased risk of HCC (OR=1.74, 95%CI=1.06-2.74). The XPD 751Gln/Gln and Gln allele genotype were associated with strong elevated susceptibility to HCC (OR=3.51 and 1.42, respectively). Conclusion: These results suggest that polymorphisms in XRCC1 and XPD may have functional significance in risk of HCC.

• 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.