• 생명과학회지
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• 제28권5호
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• pp.532-539
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• 2018

백모 발생은 연령이 증가함에 따라 모낭에 있는 melanocyte 수가 감소됨과 동시에 세포활성이 저하되고 동시에 과산화수소($H_2O_2$)가 축적되어 melanin 합성이 감소되어 진행된다. 본 연구의 목적은 흑미 주정추출물인 Black oryzasativa ethanolic extract (BLEE)가 백모에서 흑모로 전환되는데 관련되어 있는 melanin 합성에 어떠한 영향을 미치는지 알아보고자 하였다. 본 연구에서 사용된 BLEE는 DPPH radical scavenging assay 및 환원력 실험에서 $64{\mu}g/ml$에서 항산화 효과가 있는 것으로 나타났으며, tyrosinase 활성 및 melanin 생성 촉진효과는 $16{\mu}g/ml$ 이상에서 효과가 나타났다. DCF fluorescence assay에서 세포내 $H_2O_2$를 소거하는 효과가 있는 것으로 나타났고, $H_2O_2$를 처리한 세포에서 melanin 생성 촉진효과가 있는 것으로 나타났다. 그러나 Western blot 분석에서는 catalase의 발현을 감소시키는 것으로 나타났으며 melanin 생성에 관여하는 tyrosinase, TRP-1, TRP-2, 효소 및 MITF 전사인자의 발현촉진에는 영향이 없는 것으로 나타났다. 그러므로 BLEE의 melanin 생성 촉진효과는 tyrosinase 활성 증가와 세포내 $H_2O_2$의 생성을 감소시켜 melanin 생성을 촉진시키는 것으로 나타났다. 결론적으로 BLEE는 melanin 생성을 촉진시키는데 있어서 중요한 역할을 하고 있는 것으로 확인되었으며, 본 연구에서 개발된 BLEE는 흑모성장과 연관이 있는 melanin생성 촉진과 관련된 모발화장품 개발분야에서 하나의 잠재적인 기능성 소재로 응용 될 수 있다는 것을 시사하고 있다.

• Journal of Microbiology and Biotechnology
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• 제28권3호
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• pp.375-380
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• 2018

We have previously found that mycelia culture broth of eight kinds of traditional herbal extracts fermented with Phellinus linteus (previously named as 8-HsPLCB) not only inhibited melanin and tyrosinase activity, but also reduced the contents of melanogenesis-related proteins, including tyrosinase and microphthalmia-associated transcription factor, in 3-isobutyl-1-methylxanthine-stimulated B16F0 melanoma cells. For a further study, the effect of 8-HsPLCB against skin pigmentation in brown guinea pigs with ultraviolet B (UVB)-induced hyperpigmentation was investigated. 8-HsPLCB (3%) and arbutin (2%) as positive controls were applied topically twice daily for 4 weeks to the hyperpigmented areas. 8-HsPLCB showed skin-lightening effect as effective as arbutin, one of the most widely used in whitening cosmetics. Melanin index values as the degree of pigmentation showed a significant reduction week by week post 8-HsPLCB treatment and then substantially reduced by 4 weeks. The degree of depigmentation after 4 weeks of topical application with 8-HsPLCB was 32.2% as compared with before treatment (0 week). Moreover, using Fontana-Masson staining and hematoxylin-eosin staining, 8-HsPLCB reduced melanin pigmentation in the basal layer of the epidermis and epidermal thickness changes exposed to the UV-B irradiation as compared with non-treatment and vehicle treatment. The intensity of the skin-lightening effect of 8-HsPLCB was similar to arbutin. These results suggest that the skin-lightening effect of 8-HsPLCB might be resulted from inhibition of melanin synthesis by tyrosinase in melanocytes. To conclude, 8-HsPLCB treatment showed reduction of the melanin pigment and histological changes induced by UV irradiation in brown guinea pigs.

• 생명과학회지
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• 제27권12호
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• pp.1452-1461
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• 2017

백발화과정은 인간의 노화의 표현형 중 하나로 노화의 지표이다. 시간이 지남에 따라 사람 모낭에 melanocyte의 melanin 생성이 줄어들게 된다. 이의 원인으로는 모낭내 tyrosinase 활성의 감소와 활성 산소 종 중 $H_2O_2$에 의한 축적을 통해 나타난다. 천연물중에서 예로부터 흑모를 유도한다는 흑임자주정추출물의 비극성 층과 극성 층을 분리하여 항산화 효과 및 B16F1에서 melanin 생성 촉진 효과를 조사하였다. 항산화 실험에서 SBEEP와 SBEEO는 DPPH 소거 효과를 보여주지 않았고, 낮은 환원력을 보여주었다. SBEEP는 $8{\mu}g/ml$ 이상에서 세포 독성이 나타났고 SBEEO는 $4{\mu}g/ml$ 이상에서 세포 독성 효과를 보여주었다. SBEEP와 SBEEO는 in vitro에서 tyrosinase 활성과 DOPA 산화 활성에 대한 높은 melanin 합성 효과를 보여주었고, 살아있는 세포에서는 SBEEP처리군은 SBEEO 처리군보다 높은 melanin 합성 촉진 효과가 나타났다. 세포 내 melanin 생성에 효과가 있는 SBEEP는 melanin 합성 기전에서 중요한 효소인 TRP-2의 발현을 통해 melanin 생성을 촉진한다는 것을 발견하였다. 이러한 결과들은 흑임자의 SBEEP가 melanin 합성을 촉진할 수 있다는 가능성을 시사하고 있다.

• Journal of Microbiology and Biotechnology
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• 제30권8호
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• pp.1184-1194
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• 2020

Melanin is a major factor that darkens skin color as one of the defense systems to prevent the harmful effects of UV light. However, darkened skin from the localized or systemic accumulation of melanin is viewed in many cultures as an esthetic problem. Consequentially, searching for anti-melanogenic agents from natural sources is very popular worldwide. Previous screening of fermented rice products, obtained from various rice cultivars fermented with different sources of loog-pang (Thai traditional fermentation starter), revealed that the highest ability to reduce the melanin content in B16F10 melanoma cells was from unpolished black rice fermented with a defined starter mixture of microbes isolated from loog-pang E11. The aim of this study was to investigate the mechanism of the fermented unpolished black rice (FUBR) on the inhibition of melanogenesis in B16F10 melanoma cells. The strongest reduction of cellular melanin content was found in the FUBR sap (FUBRS). The melanin reduction activity was consistent with the significant decrease in the intracellular tyrosinase activity. The FUBRS showed no cytotoxic effect to B16F10 melanoma or Hs68 human fibroblast cell lines. It also significantly reduced the transcript and protein expression levels of tyrosinase, tyrosinase-related protein 1 (TYRP-1), TYRP-2, and microphthalmia-associated transcription factor. Furthermore, it induced a significantly increased level of phosphorylated ERK, p38 and Akt signaling pathways, which likely contributed to the negative regulation of melanogenesis. From these results, a model for the mechanism of FUBRS on melanogenesis inhibition was proposed. Moreover, these results strongly suggested that FUBRS possesses anti-melanogenesis activity with high potential for cosmeceutical application as a skin depigmenting agent.

• 약학회지
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• 제57권1호
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• pp.24-31
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• 2013

Although statins, inhibitors of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase, have been shown to increase melanin synthesis, the exact mechanism of this action is not fully understood. In this study we investigated the possible involvement of intracellular $Ca^{2+}$ signal in the mechanism of stimulation of melanin synthesis induced by lovastatin in B16 cells. Lovastatin stimulated the production of melanin in a dose-dependent manner in the cells. Treatment with mevalonate, FPP and GGPP, precursors of cholesterol, did not significantly suppress the lovastatin-induced melanin production, suggesting that inhibition of cholesterol synthesis may not be involved in the mechanism of the action of lovastatin. In addition, lovastatin did not significantly alter the cAMP concentration and the stimulated production of melanin by lovastatin was not significantly changed by treatment with H89, a potent inhibitor of protein kinase A, which demonstrates that cAMP pathway may not be involved. However, lovastatin increased intracellular $Ca^{2+}$ concentration in a dose-related fashion. Treatment with EGTA, an extracellular $Ca^{2+}$ chelator did not significantly alter the lovastatin-induced intracellular $Ca^{2+}$ increase and melanin synthesis, whereas intracellular $Ca^{2+}$ reduction with BAPTA/AM and intracellular $Ca^{2+}$ release blockers (dantrolene and TMB-8) completely blunted these actions of lovastatin. Taken together, these results suggest that the intracellular $Ca^{2+}$ release may play an important role in the lovastatin-induced stimulation of melanin synthesis in B16 cells. These results further suggest that lovastatin may be useful for the treatment of hypopigmentation disorders, such as vitiligo.

• 생명과학회지
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• 제34권4호
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• pp.236-244
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• 2024

Melanin은 대부분의 식물과 동물에서 발견되는 자연 색소로, 피부와 모발의 색을 결정하는 게 관여한다. Melanogenesis는 일반적으로 피부를 자외선과 같은 외부 자극으로부터 보호하기 위한 반응으로 melanocyte에서 수행한다. Tyrosinase는 기질인 tyrosine을 melanin으로 생합성 하는데 관여한다. 그러나 melanin의 과다 생성은 melasma, blotch, hyperpigmentation, 그리고 피부암과 같은 피부질환으로 이어질 수 있다. Kojic acid 및 arbutin과 같은 미백 물질에 대한 연구가 수행되었지만, 부작용으로 인해 일부 국가에서는 사용을 금지하거나 제한하고 있다. 따라서 본 연구에서는 기능성화장품의 새로운 미백재료로서 잠재력을 연구하기 위해 겨자무(horseradish)를 선택하였다. 더 나아가 효율적인 추출을 위해 아임계수 추출을 사용하였다. 연구 결과, 아임계수 200℃ 추출물은 높은 DPPH 라디칼 소거능, 총 페놀 함량, tyrosinase 저해능 그리고 B16-F10 melanoma 세포주의 melanin 생성 억제 효과를 나타내었다. B16-F10 melanoma 세포주에 대한 세포 독성을 조사하기 위해 MTT reduction assay 및 형태학적 변화를 관찰하였으며, 그 결과, 겨자무 methanol 추출물 및 아임계수 200℃에서는 세포독성이 확인되지 않았다. 결론적으로, 이 연구는 화장품의 천연 미백 기능성소재로서 가능성을 제안하는 바이다.

• 생명과학회지
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• 제28권8호
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• pp.908-916
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• 2018

모발의 검정색은 노화됨에 따라 백색으로 변화된다. 이러한 현상은 모낭내 tyrosinase 활성의 감소와 $H_2O_2$에 의한 축적에 의하여 기인된다. 따라서 본 연구의 목적은 현미주정추출물(OREE)를 이용하여 백발화의 원인인 과산화 수소에 대한 항산화 활성과 멜라닌 생성 촉진 효과를 조사한 것이다. 본 연구에서 OREE은 낮은 DPPH radical 소거능과 환원력을 보여주었다. 그러나 B16F1 세포내에서 $H_2O_2$ 소거에 대해서는 높은 항산화효과를 나타내었다. 그리고 OREE는 in vitro에서 DOPA 산화 활성은 나타나지 않았지만 $64{\mu}g/ml$에서 tyrosinase 활성을 증가시켰다. MTT assay에서 OREE는 $32{\mu}g/ml$ 이상의 농도에서 세포독성을 나타내었다. 또한 OREE는 $8{\mu}g/ml$ 이상의 농도에서 멜라닌 합성은 농도에 비례하여 증가하였고, $H_2O_2$로 멜라닌 생성을 저하시킨 세포에서도 멜라닌 합성을 증가시켰다. 멜라닌 합성에 대한 OREE의 효과를 확인하기 위하여 Western blot 분석이 수행되었다. $H_2O_2$로 멜라닌 생성을 저하시킨 세포에서 멜라닌 합성 기전 관여하는 tyrosine hydroxylase와 tyrosinase-related protein-2 (TRP-2) 발현은 OREE의 존재 하에서 증가하였다. 이상의 발견들은 OREE가 멜라닌 합성을 촉진시킬 수 있어 이와 관련된 모발화장품의 개발에 적용시킬 수 있다는 것을 시사하고 있다.

• 한방안이비인후피부과학회지
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• 제21권1호
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• pp.70-82
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• 2008

Objective : The purpose of this study is to examine the effects of Baickbujasan(BB) on the skin damage and depigmentation. Method : The inhibition of tyrosinase activity, melanogenesis and cell viability in cultured B16 melanoma cells were measured. In order to test effects of reduction of melanogenesis, B16 F-10 mouse melanoma stem line was employed to extract melanin from cultured cell, where BB was added or not, and was dissolved in alkali for colorimetric analysis. Also, in order to test skin alteration in C57BL/6 after UV irradiation, the animals were grouped into a UV urradiation group and UV irradiation after BB application group. Dopa oxidase tissue staining was excuted to invesitage the change in the distribution of active melanin cell. The distribution of active melanin cell in inner skin of iNOS after damage from UVB irradiation and the manifestation condition of P53 which takes part in natural death of keratinocyte were examined. Result : The results indicate that BB has significant effects on tyrosinase activity, and melanogenesis in vivo test. BB seems to reduce C57BL/6, external dermatological damage, for instance, erythematous papule, eczema, loss of keratinocyte, reduction in pus, and relieves dermatological damages. Conclusion : BB can be applied externally for UV protection and depigmentation.

• 대한화장품학회:학술대회논문집
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• 대한화장품학회 2003년도 IFSCC Conference Proceeding Book II
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• pp.201-212
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• 2003

The heartwood extract of Artocarpus lakoocha Roxb., which contains a potent tyrosinase inhibitor oxyresveratrol, was evaluated for its melanin-reducing efficacy in both guinea pigs and human volunteers. After 4 week-daily application of the extract dissolved in propylene glycol to the back of guinea pigs, significant reduction in melanin content was detected, with the effect greater than 3% kojic acid and solvent propylene glycol (P < 0.05). The extract was subsequently tested in female volunteers (3 groups of 20 subjects) using a parallel clinical trial with self-control. The first group received the A. lakoocha solution in propylene glycol whereas the second and the third group respectively received 0.25% licorice extract and 3% kojic acid in the same solvent. The subject in each group twice daily applied the test solution on one arm whereas the remaining arm was applied with only propylene glycol (self-control) for 12 weeks. The A. lakoocha extract was found to be the most effective agent, giving the shortest onset of significant whitening after only 4 weeks of application (P < 0.05), followed by 3% kojic acid (8 weeks) and 0.25% licorice extract (10 weeks). The whitening effect also increased with time, with the highest extent observed with A. lakoocha at week 12. The in vitro antityrosinase activity of A. lakoocha extract decreased with time upon storage at room temperature but could be stabilized by a combination of several antioxidants. In conclusion, the heartwood extract of A. lakoocha appeared to have promising potential for use as an effective and economical skin-whitening agent.

• The Plant Pathology Journal
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• 제16권3호
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• pp.125-129
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• 2000

A rapid and efficient assay to determine melanin biosynthesis inhibition of Magnaporthe grisea, a causal agent of the rice blast, by chemicals was developed. Wells in 24-well plates were loaded with spore suspension of the fungus and three known melanin biosynthesis inhibitors of KC10017, tricyclazole, and carpropamid. Subsequent color changes of mycelia and culture media in the wells were observed 7 days after incubation. The wells treated with KC10017 (an inhibitor of polyketide synthesis step and/or pentaketide cyclization step) became colorless, whereas tricyclazole (an inhibitor of 1, 3, 8-trihydroxynaphthalene reductase) or carpropamid (an inhibitor of scytalone dehydratase)-treated wells exhibited red color. They did not show any inhibitory effect on fungal growth. The inhibition of reaction steps prior to 1, 3, 6, 8-tetrahydroxynaphthalene formation was easily determined by colorless medium and mycelia. However, it was impossible to distinguish between inhibition of reduction steps and inhibition of dehydration steps by colors of the cultures. It was accomplished through HPLC analysis of the melanin biosynthesis-involving pentaketide metabolites accumulated by the inhibitors. Through screening of a number of synthetic chemicals using the in vitro assay, we could find a novel chemical group of melanin biosynthesis inhibitor.