• 대한한방부인과학회지
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• 제21권2호
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• pp.59-75
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• 2008

Purpose: This study was performed to determine the inhibitory effect of Polygonum multiflorum(PM) on melanin synthesis in B16F10. Methods: The Inhibitory effects of Polygonum multiflorum(PM) on melanin synthesis were determined by in-vitro assay. To elucidate inhibitory effects of Polygonum multiflorum on melanin synthesis, we determined the melanin release and melanin production in B16F10. And to investigate the action mechanism, we assessed the gene expression of tyrosinase, TRP-1, TRP-2, MMP-2, PKA, PKC, ERK-1 ERK-2, AKT-1, MITF in B16F10. Results: 1. PM inhibited melanin-release, melanin production in B16F10. 2. PM inhibited tyrosinase activity in vitro and in B16F10. 3. PM suppressed the expression of tyrosinase, TRP-1 in B16F10. 4. PM suppressed the expression of PKA in B16F10. 5. PM suppressed the expression of ERK-1, ERK-2, AKT-1 in B16F10. 6. PM suppressed the expression of MITF in B16F10. Conclusion: From these results, it may be concluded that PM possesses the antimelanogenetic effects.

• 대한한의학방제학회지
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• 제28권1호
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• pp.81-90
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• 2020

Objectives : This study was conducted to investigate the effects of major constituents of Epimedium koreanum Nakai (Icariin, epimedium A, epimedium B, and epimedium C) on melanin synthesis. Methods : We measured melanin contents, tyrosinase activity, and expression of Rab27a in B16F10 cells cultured with Epimedium koreanum Nakai ethanol extract (EKN) and their major constituents. After treatment with H89 and dibutyryl cAMP, which inhibit or promote the activation of PKA, we observed changes in melanin synthesis and tyrosinase activity stimulated by EKN. Results : Among them, EKN and icariin enhanced tyrosinase activity and melanin contents. We confirmed that EKN augmented melanin synthesis via cAMP/PKA pathway. Icariin-induced tyrosinase activity and melanin content were attenuated by PKA inhibitor H89, while melanogenic effect of icariin was further augmented by cAMP analog, dbc AMP. However, icariin did not affect the expression of small GTPase Rab27a involved in melanosome transport. Conclusions : These results suggest that icariin promotes melanogenesis through cAMP/PKA pathway but does not affect small GTPase Rab27a.

• Journal of Microbiology and Biotechnology
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• 제20권11호
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• pp.1513-1520
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• 2010

A bacterium capable of producing melanin pigment in the presence of L-tyrosine was isolated from a crop field soil sample and identified as Klebsiella sp. GSK based on morphological, biochemical, and 16S rDNA sequencing. The polymerization of this pigment occurs outside the cell wall, which has a granular structure as melanin ghosts. Chemical characterization of the pigment particles showed then to be acid resistant, alkali soluble, and insoluble in most of the organic solvents and water. The pigment got bleached when subjected to the action of oxidants as well as reductants. This pigment was precipitated with $FeCl_3$, ammoniacal silver nitrate, and potassium ferricynide. The pigment showed high absorbance in the UV region and decreased absorbance when shifted towards the visible region. The melanin pigment was further charecterized by FT-IR and EPR spectroscopies. A key enzyme, 4-hydroxyphenylacetic acid hydroxylase, that catalyzes the formation of melanin pigment by hydroxylation of L-tyrosine was detected in this bacterium. Inhibition studies with specific inhibitors, kojic acid and KCN, proved that melanin is synthesized by the DOPA-melanin pathway.

• 한국염색가공학회지
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• 제22권1호
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• pp.14-20
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• 2010

Squid ink was purified to melanin powder by repeated treatments with aqueous sodium hydroxide and acetic acid solutions. The exhaustion dyeing conditions of melanin to wool fabrics were investigated in relation with pH, melanin concentration, dyeing temperature and time. The melanin was dyeable to cotton and wool fabrics but higher dyeability of the wool was observed. A K/S of 7 was obtained on the optimally dyed wool fabrics with 5 % owf melanin under pH 4 at $100^{\circ}C$ for 60 minutes. Color fastness to both washing and rubbing was excellent and color fastness to light was also very good probably due to the polymeric nature of the extracted sepia melanin.

• 대한한의학회지
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• 제23권4호
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• pp.45-54
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• 2002

Objectives: In order to investigate the relationship of Radix Trichosanthis components and the melanin synthesis, the author has analyzed the cell viability and tyrosinase activity, melanin content and morphologic changes in n-hexane, EtOAc, n-BuOH, and H2O fraction. Methods: At first, in order to determine the concentration of the Radix Trichosanthis component, the author investigated the viability of B16 melanoma cell. To measure the effects of Trichosanthes kirilowii extracts (n-BuOH, n-Hexane, EtOAc, H2O fractions) on the viability of A549 cells, A549 cells were treated with various concentrations (from 0.5 to $25{\;}\mu\textrm{g}/ml$) of components of Trichosanthes kirilowii. After 24hrs, the cell viability was measured by MTT assay. The EtOAc components of Trichosanthes kirilowii decreased the viability of A549 cells in a dose-dependent manner. H2O and n-BuOH components had no cell toxicity till $25{\;}\mu\textrm{g}/ml$, the n-hexane component showed minor cell toxicity at $25{\;}\mu\textrm{g}/ml$ and the EtOAc component cell toxicity was revealed at $5{\;}\mu\textrm{g}/ml$ concentration. Results: 1. The results of tyrosinase activity and the Radix Trichosanthis component; n-hexane and EtOAc components controlled it effectively; the n-BuOH components were less effective. 2. The results of melanin content analysis showed that the n-hexane and EtOAc components effectively inhibited, the n-BuOH fraction inhibited less, and H2O component didn't inhibit the terminal melanin formation. 3. In the n-BuOH and H2O component there were no changes, but in the n-hexane component the melanin content was effectively inhibited. 4. In the EtOAc fraction, although the melanin content was inhibited, the cell count was evidently suppressed, Of all of the Radix Trichosanthis components, the n-Hexane and EtOAc fractions inhibited the melanin synthesis best, but owing to its toxicity, the EtOAc components inhibited the cell count. Conclusion: The above results demonstrated that Radix Trichosanthis n-hexane fraction efficiently inhibited the tyrosinase activity and melanin synthesis.

• 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.

• Nutrition Research and Practice
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• 제11권3호
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• pp.190-197
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• 2017

BACKGROUND/OBJECTIVES: Gallus gallus domesticus (GD) is a natural mutant breed of chicken in Korea with an atypical characterization of melanin in its tissue. This study investigated the effects of melanin extracts of GD on osteoblast differentiation and inhibition of osteoclast formation. MATERIALS/METHODS: The effects of the melanin extract of GD on human osteoblast MG-63 cell differentiation were examined by evaluating cell viability, osteoblast differentiation, and expression of osteoblast-specific transcription factors such as bone morphogenetic protein 2 (BMP-2), small mothers against decapentaplegic homologs 5 (SMAD5), runt-related transcription factor 2 (RUNX2), osteocalcin and type 1 collagen (COL-1) by reverse transcription-polymerase chain reaction and western blotting analysis. We investigated the inhibitory effect of melanin on the osteoclasts formation through tartrate-resistant acid phosphatase (TRAP) activity and TRAP stains in Raw 264.7 cell. RESULTS: The melanin extract of GD was not cytotoxic to MG-63 cells at concentrations of $50-250{\mu}g/mL$. Alkaline phosphatase (ALP) activity and bone mineralization of melanin extract-treated cells increased in a dose-dependent manner from 50 to $250{\mu}g/mL$ and were 149% and 129% at $250{\mu}g/mL$ concentration, respectively (P < 0.05). The levels of BMP-2, osteocalcin, and COL-1 gene expression were significantly upregulated by 1.72-, 4.44-, and 2.12-fold in melanin-treated cells than in the control cells (P < 0.05). The levels of RUNX2 and SMAD5 proteins were higher in melanin-treated cells than in control vehicle-treated cells. The melanin extract attenuated the formation of receptor activator of nuclear factor kappa-B ligand-induced TRAP-positive multinucleated RAW 264.7 cells by 22%, and was 77% cytotoxic to RAW 264.7 macrophages at a concentration of $500{\mu}g/mL$. CONCLUSIONS: This study provides evidence that the melanin extract promoted osteoblast differentiation by activating BMP/SMADs/RUNX2 signaling and regulating transcription of osteogenic genes such as ALP, type I collagen, and osteocalcin. These results suggest that the effective osteoblastic differentiation induced by melanin extract from GD makes it potentially useful in maintaining bone health.

• 생명과학회지
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• 제27권4호
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• pp.423-429
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• 2017

모발의 색은 melanin의 종류와 양에 의해 결정 된다. Melanocyte는 주로 자외선 자극에 의하여 L-tyrosine으로부터 멜라닌을 합성한다. 활성산소종은 백발의 진행과정에 중요한 역할을 하고 있다. 적하수오는 백발화되는 노화과정을 억제한다고 보고되고 있다. 본 연구의 목적은 흑모발생과 연관된 melanin 합성에 대한 적하수오 에탄올 추출물(PMEE)의 효능을 조사한 것이다. 항산화 실험에서 PMEE는 DPPH radical을 감소시켰고 환원력을 증가시켜 PMEE는 백발과정에 관여하는 활성산소종을 소거할 수 있다고 볼 수 있다. PMEE는 농도 의존적으로 세포생존능을 감소시켰다. 더욱이 melanin의 생성에 대한 PMEE의 효과는 DOPA assay와 tyrosinase 활성측정에 의하여 결정되었다. PMEE는 tyrosinase 활성을 증가시켰으며 melanin 합성을 촉진시켰다. 뿐만 아니라 Western blot 분석을 이용하여 tyrosinase, TRP-1, TRP-2 및 MITF 뿐만 아니라 SOD-3와 catalase와 같은 항산화 효소의 단백질 발현을 조사하였다. PMEE의 항산화 활성의 증가로 인하여 SOD-3와 catalase의 발현 수준은 감소된 것으로 나타났다. 특히, PMEE는 tyrosinase와 TRP-2의 발현 수준은 증가시켰다. 이러한 결과들은 PMEE가 백발화 된 모발에서 흑발로 변하는 것과 관련된 melanin 합성을 촉진할 수 있다는 것을 암시하고 있다.

• 생명과학회지
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• 제29권4호
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• pp.461-469
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• 2019

본 연구의 목적은 흑모 성장과 관련 있는 천연물을 개발하기 위하여 흑미, 흑임자, 현미, 서목태 및 적하수오의 에탄올 추출혼합물의 melanin 합성에 대한 효능을 조사한 것이다. 흑모가 백모로 변화되는 원인으로 $H_2O_2$가 모낭 주위에 축적되어 melanin 합성이 저하되어 백모가 유발되는 것으로 보고된 바 있다. MIXEE는 DPPH radical에 대한 소거능력과 활성산소에 대한 환원력이 있는 것으로 관찰되었다. 뿐만 아니라 세포내 $H_2O_2$ 수준을 감소시켰는데, 이러한 결과는 MIXEE는 산화를 억제하여 $H_2O_2$에 의하여 감소된 melanin 합성을 촉진 시킬수 있다는 것을 나타내었다. 한편 in vitro에서 MIXEE는 tyrosinase 활성에는 영향을 주지 않았지만 L-dopa에서 melanin 합성을 촉진하였다. 더욱이, B16F1세포에서 MIXEE는 melanin 생성을 세포수준에서 촉진하였다. Western blot을 이용한 단백질 발현 분석에서 MIXEE는 SOD-2 및 SOD-3. 특히 DOPA에서 melanin 이 생성되는 과정에 관여하는 효소인 TRP-2 발현에 긍정적인 영향을 미치는 것으로 입증되었다. 이상의 결과는 MIXEE가 백발화과정을 가역화시키는 과정에서 주요한 역할을 하는 melanin 생성을 촉진하는 하나의 약학 소재로 응용될 수 있다는 것을 시사하고 있다.

• Environmental Analysis Health and Toxicology
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• 제31권
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• pp.9.1-9.5
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• 2016

Objectives There have been developed to use targeting ability for antimicrobial, anticancerous, gene therapy and cosmetics through analysis of various membrane proteins isolated from cell organelles. Methods It was examined about the lysosomal membrane protein extracted from lysosome isolated from HeLa cell treated by 100 ppm melanin for 24 hours in order to find associated with targeting ability to melanin using by 2-dimensional electrophoresis. Results The result showed 14 up-regulated (1.5-fold) and 13 down-regulated (2.0-fold) spots in relation to melanin exposure. Conclusions It has been found that lysosomal membrane proteins are associated with melanin to decolorize and quantity through cellular activation of lysosome.