• 약학회지
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• 제47권1호
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• pp.31-36
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• 2003

To investigate the effect of protein kinase on melanin production via cAMP-dependent pathway, we measured the melanin amount and tyrosinase activity in B16 melanoma cells stimulated by alpha-melanocyte stimulating hormone (MSH), forskolin and 8-Br-cAMP. MSH, forskolin and 8-Br-cAMP significantly increased both melanin production and tyrosinase activity in B16 cells. Melanin production and tyrosinase activity by MSH are significantly inhibited by cyclic AMP-dependent protein kinase inhibitor (KT5720) and protein kinase C down-regulation treated with PMA. Bisindolmaleimide (1$\mu$M), protein kinase C inhibitor, significantly inhibited melanin production and tyrosinase activity stimulated by MSH, forskolin and 8-Br-cAMP with the following order of potency: MSH＞forskolin＞8-Br-cAMP. Tyrosine kinase inhibitor, genistein and DHC, significantly inhibited both, but the inhibitory effect was more potent in 8-Br-cAMP-stimulated B16 cells than MSH-stimulated cells. NFkB inhibitor (parthenolide) significantly inhibited melanin production and tyrosinase activity. Neither melanin production nor tyrosinase activity induced by MSH, forskolin and 8-Br-cAMP were affected by KN-62 (calmodulin-dependent protein kinase II inhibitor), PD098059 (mitogen-activated protein kinase inhibitor, MAPKK) and worthmannin (phosphatidylinositol 3-kinase inhibitor). These results suggest that both protein kinase C and tyrosine kinase are involved in melanin production by cyclic AMP-dependent pathway and NFkB pathway may play an important role in cyclic AMP-dependent melanin production in B16 melanoma cells.

• 대한화장품학회지
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• 제23권3호
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• pp.101-107
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• 1997

Phellinus linteus was artificially cultivated in kangwon province in Korea. The air-dried phellinus linteus was frozen in liquid nitrogen tank and powdered in jar. 10g of the powder was extracted with each 200g of ethanol, methanol, distilled water and 1,3-butylene glycol/distilled water 4 hours under refluxing and then the liquidextract was concentrated under reduced pressure. As a result of analysis by high performance liquid chromatography and thin layer chromarography, many kinds of sugar and flavonoids were detected. Also we knew that phellinus linteus' extract had a strong UV-ray absorption. In the efficacy test for applying to cosmetics, free radical scavenging effect was confirmed. As a result, 2% of sample was the most potent inhibitory effect and the free radical savenging activity, was 0.31%. This is more effective than any other meterial. In the test of antioxidative activity against lipid autoxidation, phellinus linteus' extract had a good effect by 46% while vitamine E was 42.3%. The immunological activity of phellinus linteus was showed through the activation of macrophage cell. Actually, phellinus linteus activated macrophage function of 1.1-1.8 times including nitrite production compared to control. The whitening effect of phellinus linteus was showed through the inhibition of tyrosinase activity, melanin biosynthesis of S. bikiniensis and B-16 melanoma cells. Phellinus linteus' extract was showed strong mushroom tyrosinase inhibitory activity with IC50 value of 0.5% and inhibited melanin biosynthesis with 28mm inhibition zone at 0.005%/paper disc in S. bikinniensis, a bacterium used as an indicator organism in this work. Also it inhibited melanin biosynthesis in B-16 melanoma cells with a minimum inhibitory concentration of 0.134%.

• 대한본초학회지
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• 제34권6호
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• pp.109-115
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• 2019

Objective : Herbal medicinal mixture (JMB) are consisted of Caryophylli Flos, Aucklandiae Radix, and Angelicae Dahuricae Radix. Each of these herbal medicines has studied on anti-aging effect in vitro. So this study was conducted to investigate efficacy and potency of JMB extract on dermal anti-aging and whitening. Methods : The JMB was extracted at room temperature by 80% ethanol. Collagenase and elastase inhibition activity in JMB ethanol extract were determined at 10, 50, 100, 500, 1000 mg/ml concentrations by colorimetric method. The toxic range of JMB ethanol extract was evaluated using MTT assay. Also, The inhibitory effect of JMB ethanol extract on tyrosinase activity and melanin contents in mouse melanoma cell line (B16F10 cell) was identified at 50, 100, 200 ㎍/㎖ levels by spectrometric assay. In each analysis, EGCG (epigallocatechin gallate) and Kojic acid were used as positive controls, respectively. Results : The elastase and collagenase inhibitory activity of JMB ethanol extract increased dose dependently. Also, The MTT assay showed that JMB, up to 400 ㎍/㎖ concentration, exhibited no toxic effect to the B16F10 cell. And following the JMB ethanol extract treat, cellular melanin contents and tyrosinase activity were dose-dependently decreased compared to those of control. Conclusion : These results suggest that JMB ethanol extract has effects to inhibitory activity on dermal wrinkle enzyme and melanogenesis. Therefore, JMB has applicable benefits for development of materials or products to have whitening and anti-aging functions on skin.

• 대한화장품학회지
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• 제32권1호
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• pp.23-28
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• 2006

본 연구는 택사의 화장품 원료로서의 특성을 알아보기 위하여 화장품의 기능들인 항산화, 미백, 세포 손상 회복 및 항염증과 관련된 다양한 실험을 실시하였다. 30, 70, 100% MeOH 용매로 추출한 택사 추출물들은 DPPH법으로 실시한 free radical scavenging assay에서 좋은 활성을 보여주었고, tyrosinase 활성 억제 시험에서도 0.5% 이상의 농도에서 농도 의존적인 활성을 보여주었다. Human fibroblast를 사용한 proliferation assay (MTT assay)에서 각 용매 추출물들은 별다른 효과를 보여주지 못했고 0.05% 이하의 농도에서는 세포 독성으로부터 안전하다는 것을 알 수 있었다. Bl6 melanocyte를 사용한 melanin 생성 억제시험에서 각 용매별 추출물은 독성으로부터 안전한 0.05% 이하의 농도에서 melanin 생성을 40% 이상 억제하는 높은 활성을 보여주었다. 이후 우리는 택사 추출물의 MPLC분리 분획을 실시하여 세 가지 분획을 얻었으며 이들을 대상으로 세포 손상 회복시험과 melanin생성 억제 시험, 염증인자 생성 억제 시험을 실시하였다. 그 결과 분획물들 중 3번 분획물이 세포 손상 회복을 30% 이상 올려주는 좋은 결과를 보여주었고, melanin 생성 억제 시험과 COX-2 생성억제에서도 주목할만한 결과를 보여주었다.

• 대한한방피부미용학회지
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• 제1권1호
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• pp.5-15
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• 2005

The purpose of this study was to investigate the effect of the aqueous extract of the Astragalus membranaceous(AM). AM showed inhibitory effect on the tyrosinase activity using L-tyrosine as a substrate. Tyrosinase plays an important role in the process of melanin polymer biosynthesis. In vitro AM extract(1mg/ml) inhibited melanin biosynthesis and are useful for the material used in cosmetics. B16 mouse melanoma cells were cultured in different concentrations. The non-cytotoxicity of the plant extracts was confirmed by MTT assay.

• Korean Journal of Acupuncture
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• 제37권1호
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• pp.24-30
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• 2020

Objectives : The purpose of this study was to investigate melanogenesis inhibition of ethanol extract of Polyporus (EP) by using B16F10 melanoma cells. Methods : We measured antioxidant effect of EP by using 1,1-Diphenyl-1-picrylhydrazyl (DPPH) assay and we confirmed melanin contents and tyrosinase activity of EP in cells. Additionally, the expression of tyrosinase-related protein-1 (TRP-1) and TRP-2 was observed by Western blot. Results : EP showed significantly high radical scavenging activity and inhibition of melanogenesis in dose-dependent manner by decreasing cellular tyrosinase activity and melanin content with or without α-melanin stimulating hormone. TRP-1 and TRP-2 expressions were also suppressed by EP in B16F10 cells. Conclusions : These results suggest that EP inhibits the melanogenesis and it could be a new organic ingredient for hyper-pigmentation.

• 한방안이비인후피부과학회지
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• 제16권1호
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• pp.100-111
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• 2003

Recently many efforts were focused to understand the mechanical insights of melanogenesis to develop the agents for hyper-pigmentation and hypo-pigmentation. In the melanin biosynthetic pathway, tyrosinase is the rate limiting enzyme, and ${\alpha}$-melanocyte stimulating hormone(MSH) or cAMP-elevating agents stimulate melanogenesis and enhance the melanin synthesis and the tyrosinase activity. The author has analyzed the effects of Rhizoma Bletillae on the basal melanogenic activities of B16 mouse melanoma cells. Rhizoma Bletillae alone markedly suppressed melanin content and tyrosinase activity in a dose-dependent manner. Pretreatment of the cells with Rhizoma Bletillae. The decrease in the tyrosinase activity was paralled by a decrease in the abundance of tyrosinase protein and tyrosinase promoter activity. These results suggest that Rhizoma Bletillae inhibits melanogenesis of B16 melanoma cells via suppression of tyrosinase activity.

• 대한화장품학회지
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• 제34권2호
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• pp.109-115
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• 2008

기질 화합물로써 $3{\beta}$-hydroxy-12-oleanen-28-oic acid 유도체들의 치환기($R_1{\sim}R_4$)가 변화함에 따른 protein tyrosine phosphatase(PTP)-1B의 저해활성에 관한 비교 분자장 분석(CoMFA) 모델을 유도하고 정량적으로 검토하였다. 최적의 CoMFA F1 모델은 가장 높은 예측성과 상관성($r^2_{cv.}=0.654$ 및 $r^2_{ncv.}=0.995$)을 나타내었다. 저해활성에 관한 CoMFA장 기여비율(%)의 순서는 입체장(53.0%), 정전기장(36.2%) 및 소수성장(10.8%) 이었다. 등고도 분석 결과로부터 저해활성은 기질 분자 내 $R_4$-치환기에 의존적이었으며 특히 melanin 저해활성이 높은 새로운 화합물(P1 및 P2)이 예측되었다.

• 한국잠사곤충학회지
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• 제51권2호
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• pp.207-210
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• 2013

본 연구는 누에 숙잠 혈림프의 멜라닌 생성 억제 효과에 대해 알아보기 위해 수행되었다. 실샘을 제거한 누에 숙잠에서 혈림프를 정제하였고, 이의 항산화, 멜라닌 생성 억제, 티로시나아제 활성 억제 효과 여부를 실험한 결과는 다음과 같다. 1. 숙잠의 실샘을 제거한 후, 나머지 누에조직을 분쇄하여 원심분리 후 여과하였던 기존의 방식에서 연속식 원심분리로 정제 방법을 개선한 결과, 수율은 53% 증가하였고 정제기간은 5일 단축되었다. 2. 누에 숙잠 혈림프의 DPPH 자유라디칼 소거능을 측정한 결과 비타민 C 보다는 효과가 떨어졌지만 세리신 단일 물질보다 높은 효과를 나타내었다. 3. 숙잠 혈림프 단백질을 음이온 교환 크로마토그래피로 분리하여 활성산소 억제 효과를 측정한 결과, 70 kDa과 30 kDa 단백질이 모두 포함되어 있는 혈림프 단백질(상등액)이 대조군보다 2배, 70 kDa 단백질이 30 kDa 단백질에 비해 약 1.7배 활성산소 억제 효과를 나타냄을 확인하였다. 4. 누에 숙잠 혈림프의 멜라닌 생성 억제 효과는 미백화장품 소재로 널리 사용되는 상백피와 알부틴보다 각각 9.15%, 11.56% 더 우수함을 확인하였다(혈림프 1% 농도 기준). 5. 티로시나아제 활성 억제 효과는 대조인 상백피보다 누에 숙잠 혈림프의 티로시나아제 활성 억제 효과가 우수하였으나 단일 물질인 알부틴보다는 억제 효과가 떨어지는 결과를 나타내었다. 하지만 정제 혈림프 농도 $450{\mu}g/m{\ell}$에서는 90%에 이르는 티로시나아제 활성 억제 효과를 확인하였다.

• 분석과학
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• 제19권3호
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• pp.194-202
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• 2006

중국산 천연물질인 서장채국화, 홍경천 및 장청과에서 미백성분을 추출 및 정제하였다. 잘게 분쇄한 서장채국화와 장청과 잎을 각각 물과 에틸 에테르로 추출하고 제조용 크로마토그래피 GS310 컬럼($21.5{\times}500mm$ i.d., $10-15{\mu}m$)으로 네 개 혹은 세 개의 부분으로 획득한 후 농축하였다. 잘게 분쇄한 홍경천은 메탄올로 추출하고 분석용 $C_{18}$ 컬럼 ($3.9{\times}300mm$ i.d., $15{\mu}m$)으로 2개의 부분으로 획득한 후 농축하였다. 미백효과는 in-vitro 멜라닌 생성효과로 측정하였다. 이때 사용한 멜라닌-a와 B16 셀의 농도는 $10{\mu}g/ml$이다. $200{\mu}g/ml$의 농도에서 서장채국화의 에틸 아세테이트층은 92%의 멜라닌 억제효과를 보이며, 홍경천은 60%의 멜라닌 억제효과를 보였다. 이것은 모두 알부틴의 미백효과(45.6%)보다 우수하였다. $100{\mu}g/ml$의 농도에서 장청과는 90%이상의 멜라닌 억제효과를 나타냈지만 B16흑색종 셀에 대하여 독성을 보였다.