• Archives of Pharmacal Research
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• 제27권12호
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• pp.1226-1232
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• 2004

Extracts from seventeen seaweeds were determined for tyrosinase inhibitory activity using mushroom tyrosinase with L-tyrosine as a substrate. Only one of them, Ecklonia stolonifera OKAMURA (Laminariaceae) belonging to brown algae, showed high tyrosinase inhibitory activity. Bioassay-guided fractionation of the active ethyl acetate (EtOAc) soluble fraction from the methanolic extract of E. stolonifera, led us to the isolation of phloroglucinol derivatives [phloroglucinol (1), eckstolonol (2), eckol (3), phlorofucofuroeckol A (4), and dieckol (5)]. Compounds 1~5 were found to inhibit the oxidation of L-tyrosine catalyzed by mushroom tyrosinase with $IC_{50}$ values of 92.8, 126, 33.2, 177, and 2.16 ${\mu}g$ /mL, respectively. It was compared with those of kojic acid and arbutin, well-known tyrosinase inhibitors, with $IC_{50}$ values of 6.32 and 112 ${\mu}g$ / mL, respectively. The inhibitory kinetics analyzed from Lineweaver-Burk plots, showed compounds 1 and 2 to be competitive inhibitors with $K_i$ of $2.3{\times}10^{-4}\;and\;3.1{times}10^{-4}$ M, and compounds 3~5 to be noncompetitive inhibitors with $K_i$ of $1.9{\times}10^{-5},\;1.4{\times}10^{-3}\;and\;1.5{\times}10^{-5}$ M, respectively. This work showed that phloroglucinol derivatives, natural compounds found in brown algae, could be involved in the control of pigmentation in plants and other organisms through inhibition of tyrosinase activity using L-tyrosine as a substrate.

• Archives of Pharmacal Research
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• 제27권2호
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• pp.169-172
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• 2004

Activity-guided fractionation of the EtOAc-soluble extract of the stems of Couepia ulei, using a bioassay based on the induction of quinone reductase (QR) in cultured Hepa 1c1c7 mouse hepatoma cells led to the isolation of two active compounds, a new natural product, erythro-2,3-bis(4-hydroxy-3-methoxyphenyl)-3-ethoxypropan-1-o1 (1), and a known compound, evofolin-B (2), along with five inactive compounds all of known structure, viz., betulinic acid, oleanolic acid, pomolic acid, ($\pm$)-syringaresinol, and ursolic acid. These isolates were identified by analysis of physical and spectral data. Compounds 1 and 2 exhibited QR inducing activity, with observed CD (concentration required to double induction) values of 16.7 and 16.4 $\mu\textrm{M}$, respectively.

• 임산에너지
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• 제19권1호
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• pp.49-60
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• 2000

본 실험에서는 3종의 한국산 겨우살이 메탄올 추출물을 유기용매 분획과 컬럼크로마토그래피를 이용한 분획을 실시한 후, Tyromyces palustris, Endothia nischkei 그리고 Trichophyton rubrum에 대한 항균활성을 평가하여 겨우살이 추출물의 항균활성 자료로 활용하고자 하였다. 그 결과 모든 겨우살이 추출물은 갈색 불후균에 대한 활성은 매우 낮게 나타났다. 그러나 Viscum albumvar. coloratum 추출물중 부탄올 분획은 황색줄기 마름 병원균의 성장을 억제하는 것으로 나타났으며, 이 겨우살이의 EtOAC 분획은 족부백선균에 대하여 높은 포자 발아 억제효과를 나타내었다. 시험된 3종의 한국산 경우살이류 중 V. album var. Coloratum은 병원균에 대하여 황균활성 성분을 함유할 가능성이 가장 높을 것으로 제시되었다.

• 한국식품과학회지
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• 제47권2호
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• pp.170-175
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• 2015

본 연구에서는 국산배의 기능학적 우수성 증명을 위해 국산배로부터 생리활성 화합물을 밝히고자 하였다. 이에 배 과피 MeOH 추출물을 용매분획하여 얻은 EtOAc-산성획분을 대상으로 Sephadex LH-20 column chromatography와 ODS-HPLC를 이용하여 순차적으로 정제를 행하여 2종의 화합물을 단리하였다. 단리된 화합물 1과 2는 MS 및 NMR 분석을 통하여 각각 quercetin 3-O-${\beta}$-Dglucopyranoside(화합물 1)와 3,5,6,7,8,3',4'-heptahydroxyflavan [(-)-dulcisflavan, 화합물 2)]으로 동정되었다. 단리된 화합물 1은 동양배로부터, 그리고 화합물 2는 배로부터 처음 동정된 화합물이다. 본 연구결과가 국산배의 기능학적 우수성 증명을 위한 기초자료로 활용되길 기대한다.

• 한국식품영양학회지
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• 제18권4호
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• pp.367-372
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• 2005

밤나무 잎에 함유된 기능성 물질 검색의 일환으로 항미생물 효과를 검정하고 활성물질의 분석을 시도하였다. 예비실험을 통해 추출물의 활성을 확인한 후 시료를 n-hexane, EtOAc, MeOH로 순차 추출하여 정제한 다음 활성이 강한 MeOH 분획의 0.2 g, 0.5 g에 해당하는 추출물로 13종의 미생물에 대해 활성을 측정한 결과, Cram 양성 세균 중에서 S. aureus와 S. epidermidis는 대조구로 사용한 benzoic acid보다 강한 활성이 나타났고, M. luteus와 L. mesenteroides, B. subtilis에서도 활성이 인정되었으며, Gram 음성 세균인 E. coli, S. typhimurium, P. aeruginosa에서도 강한 활성이 나타나 밤나무 잎 추출물은 비교적 넓은 항균 spectra와 활성이 있음이 확인되었다. 이에 활성물질을 규명하고자 시료를 solvent fractionation, silica gel adsorption column chromatography, Sephadex LH-20 column chromatography, HPLC 등으로 정제하고 활성이 나타난 산성 획분과 페놀성 획분의 물질을 분리한 후 MS, $^1H-NMR,\;^{13}C-NMR$ 등으로 분석한 결과, 산성 획분에서는 stigmast- 5-en-3-ol($\beta$-sitosterol)이 활성을 갖는 물질로 나타났다.

• Nutrition Research and Practice
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• 제8권5호
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• pp.509-515
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• 2014

BACKGROUND/OBJECTIVES: The root of Vitis amurensis Ruprecht, a sort of wild-growing grape, has been used in oriental medicine for treatment of skin ailments; however, its dermatological activity is not sufficiently understood. The aim of this study was to investigate tyrosinase inhibitory and anti-melanogenic activities of V. amurensis Ruprecht root methanol extract (VARM) in B16F10 mouse melanoma cells and to attempt to isolate and identify the active compound issued from VARM. MATERIALS/METHODS: Anti-melanogenic activity of VARM was analyzed in ${\alpha}$-melanocyte stimulating hormone (MSH)-stimulated B16F10 cells through evaluation of antioxidative activity as well as inhibited tyrosinase activity and melanin contents compared with those of kojic acid and arbutin. After anti-melanogenic analysis of VARM, serial fractionation, nuclear magnetic resonance (NMR), and thin layer chromatorgraphy (TLC) were applied for identification of active compounds contained in VARM. RESULTS: VARM significantly inhibited oxidative stress and tyrosinase activity and attenuated ${\alpha}$-MSH-induced melanin production in B16F10 cells. For isolation of active compounds, VARM was fractionated using a series of organic solvents, including dichloromethane ($CH_2Cl_2$), ethyl acetate (EtOAc), and n-butanol (n-BuOH). Among fractions showing anti-melanogenic activity, the CH2Cl2 fraction induced the most potent attenuation of melanogenesis without cytotoxicity and the major compound in the $CH_2Cl_2$ fraction was identified as betulinic acid. Betulinic acid isolated from the $CH_2Cl_2$ fraction of VARM significantly attenuated ${\alpha}$-MSH-induced melanogenesis in a dose dependent manner, which was stronger than that of arbutin used as a positive control. CONCLUSIONS: These results indicate that VARM inhibits oxidative stress, tyrosinase activity, and ${\alpha}$-MSH-induced melanogenesis in B16F10 cells, due primarily to the active compound, betulinic acid, in the $CH_2Cl_2$ fraction.

• Applied Biological Chemistry
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• 제27권1호
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• pp.40-44
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• 1984

고등식물(高等植物)이 균근균(菌根菌)과 공생관계(共生關係)를 이루면 생장촉진(生長促進)의 특이현상(特異現象)을 보이는데, 이러한 생리현상(生理現象)을 구명(究明)하기 위하여 침엽수의 외생균근(外生菌根)의 주형성균(主形成菌)인 Pisolithus tinctorius가 생산(生産)하는 GA활성(活性)을 검색(檢索)하였다. P. tinctorius 배양액(培養液)에서 추출(抽出), 용매분획(溶媒分劃), Sephadex LH-20관(管)크로마토그래피, Silica gel분배(分配) 크로마토그래피, TLC에 의(依)해 분획(分劃), 정제(精製)된 추출물(抽出物)을 단은방주(短銀坊主)를 사용(使用)한 생물검정(生物檢定)에 의(依)해 GA활성(活性)을 조사한 결과 Silica gel 분배(分配) 크로마토그래피의 초산(醋酸)에칠 $30{\sim}60%$ 용출구(溶出區)와 TLC의 Rf $0.1{\sim}0.4,\;0.6{\sim}0.8$에서 GA활성(活性)을 나타냈다. GA활성(活性)은 유묘개체당(幼苗個體當) 배양추출액(培養抽出液)의 처리양을 증가하면 활성(活性)도 따라서 증가하였으며, 활성(活性)은 P. tinctorius 100ml당(當) $GA_3$ 0.1ng에 상당(相當)하였다.

• 대한한방소아과학회지
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• 제30권3호
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• pp.1-30
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• 2016

Objectives Previous studies have found out that Forsythiae Fructus (FF) extracts have anti-atopic activities by in vitro experiment. In order to understand more about FF extracts' benefit, we subdivided FF extracts depending on systematic fractionation method by using Methylene chloride (MC), Ethyl acetate (EtOAc), n-BuOH and n-hexane (n-Hx). This study is designed to examine the effect of FF fractions on the PMA- ionomycin-induced activation of RBL-2H3 mast cell lines in vitro and on the DNCB-induced activation of NC/Nga mice in vivo. Methods For this study, we examined IL-4, IL-13 production by ELISA analysis, IL-4, IL-13, IL-31, IL-31RA and TNF-${\alpha}$ mRNA expression by real-time PCR and manifestations of AP-1 and MAPKs transcription factors by western blotting in vitro. Through in vitro experiment, we selected FF n-BuOH fraction that seems the best effective in atopic dermatitis then induced it on NC/Nga mice by DNCB. We measured mice's WBC, eosinophil and neutrophil in heart blood, IL-4, IL-5, IFN-${\gamma}$ in the spleenocyte culture supernatant, the absolute cell numbers of CD4+, CD8+, B220+CD23+, CD3+CD69+ and Gr-1+CD11b+ in the PBMCs, ALN and dorsal skin, IL-5, IL-13, IL-31, IL-31RA in the dorsal skin by real-time PCR and the distribution of immune cells by H&E on dorsal skin and ANL and toluidine blue staining on dorsal skin. Results FF n-BuOH fraction suppressed IL-4, IL-13 production and mRNA expression of IL-4, IL-13, IL-31, IL-31RA and TNF-${\alpha}$. Results from the western blot analysis showed that FF n-BuOH fraction reduced the activation of the mast cell specific transduction factors involved in AP-1 by suppressing JNK and ERK phosphorylation. In the gross, atopic dermatitis induced by DNCB in NC/Nga mice were improved by oral administration of FF n-BuOH fraction. Oral FF n-BuOH fraction also decreased the level of IgE in mice's serum and the level of IL-4 and IL-5 in the spleenocyte culture supernatant, cell numbers of CD8+, B220+CD23+ in the PBMCs, CD4+ in the ALN and CD4+, Gr-1+CD11b+ in the dorsal skin and suppressed mRNA expression of IL-5, IL-13, IL-31, IL-31RA in the dorsal skin. Histological examination showed that infiltration levels of immune cells in atopic dermatitis induced NC/Nga mice were improved by FF n-BuOH fraction. Conclusions FF n-BuOH fraction can reduce pruritus by suppressing IL-31, IL-31RA secretion and modulate molecular mediators and immune cells associated with atopic dermatitis induced in NC/Nga mice which may have played a significant role in recovering atopic dermatitis symptoms.

• Archives of Pharmacal Research
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• 제26권8호
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• pp.585-590
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• 2003

Activity-guided fractionation of the EtOAc-soluble extract of the whole plants of Sida acuta using a bioassay based on the induction of quinone reductase (OR) in cultured Hepa 1c1c7 mouse hepatoma cells, led to the isolation of ten active compounds of previously known structure, quindolinone (1), cryptolepinone (2), 11-methoxyquindoline (3), N-trans-feruloyltyramine (4), vomifoliol (5), loliolide (6), 4-ketopinoresinol (7), scopoletin (8), evofolin-A (9), and evofolin-B (10), along with five inactive compounds of known structure, ferulic acid, sinapic acid, syringic acid, ($\pm$)-syringaresinol, and vanillic acid. These isolates were identified by physical and spectral data measurement. A new derivative of quindolinone, 5,10-dimethylquindolin-11-one (1a) was synthesized and characterized spectroscopically. Of the active substances, compounds 1-3 and 1a exhibited the most potent QR activity, with observed CD (concentration required to double induction) values ranging from 0.01 to 0.12 $\mu$ g/mL. Six compounds were then evaluated in a mouse mammary organ culture assay, with cryptolepinone (2), N-trans-feruloyltyramine (4), and 5,10-dimethylquindolin-11-one (1a) found to exhibit 83.3, 75.0, and 66.7% inhibition of 7,12-dimethylbenz[a]anthracene-induced preneoplastic lesions, respectively, at a dose of 10 $\mu\textrm{g}$/mL.