• Title/Summary/Keyword: Sephadex LH-20

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Differential Effects of Resveratrol and its Oligomers Isolated from Seeds of Paeonia lactiflora (Peony) on Proliferation of MCF-7 and ROS 17/2.8 Cells

  • Kim, Hyo-Jin;Lee, Won-Jung;Park, Yun-Hee;Cho, Sung-Hee;Park, Sang-Won
    • Preventive Nutrition and Food Science
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    • v.8 no.4
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    • pp.356-364
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    • 2003
  • A methanol extract from seeds of Paeonia lactiflora (Paeoniaceae, peony) was found to possess different antiproliferative activities against four different human cancer cell lines: Hela, MCF-7, HepG2 and HT-29. Furthermore, five different methanol (20, 40, 60, 80 and 100 % MeOH) fractions obtained by fractionation of the methanol extract of the seeds on a Diaion HP-20 column exhibited differential antiproliferative effects against the above four cancer cell lines. Among five fractions, the 60 % MeOH fraction showed relatively lower antiproliferative activity on MCF-7 estrogen-sensitive breast cancer cell than the other cancer cell lines. Systematic separation of 60% the MeOH fraction by silica gel and Sephadex LH-20 columns led to the isolation of four known stilbenes, trans-resveratrol (1), trans-(+)- $\varepsilon$ -viniferin (2), gnetin H (3) and suffruticosol B (4). The four stilbenes (1∼4) exerted differential biphasic effects on cell proliferation of MCF-7 cells in a similar manner as genistein, a soybean isoflavone used as a positive reference, in the concentration range from 1.0 to 200 $\mu$M. Three stilbenes (1 ∼ 3) weakly stimulated the proliferation of MCF -7 cells at doses below 10 JIM. However, strong antiproliferative effects on MCF-7 cell were exerted by extract 1 at a dose of 200 JIM, and by 2 and 3 at doses above 25 $\mu$M. In contrast, 4 inhibited the proliferation of MCF-7 cell at a dose below 25 $\mu$M, but stimulated cell proliferation at concentrations of 50 and 100 $\mu$M. All four stilbenes (1∼4) stimulated the proliferation of ROS 17/2.8 osteoblast-like cells in the range of 10$^{-10}$ ∼10$^{-1}$ $\mu$M. Compound 1 exhibited especially potent proliferative activity, although its activity was weaker than that of genistein. Additionally, three resveratrol oligomers (2∼4) also exhibited concentration-dependently moderate proliferative activity, but less than that of 1. These results suggest that resveratrol, and its dimer and trimers from the seeds of Paeonia lactiflora may act as a phytoestrogen, but in a somewhat different manner from that of genistein.

Isolation and Identification of 3 Low-molecular Compounds from Pear (Pyrus pyrifolia Nakai cv. Chuhwangbae) Fruit Peel (추황배(Pyrus pyrifolia Nakai cv. Chuhwangbae) 과피로부터 3종의 저분자 화합물의 단리·동정)

  • Lee, Yu Geon;Cho, Jeong-Yong;Kim, Chan-Mi;Jeong, Hang-Yeon;Lee, DongI;Kim, Soo Ro;Lee, Sang-Hyen;Kim, Wol-Soo;Park, Keun-Hyung;Moon, Jae-Hak
    • Korean Journal of Food Science and Technology
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    • v.45 no.2
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    • pp.174-179
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    • 2013
  • Three low-molecular compounds were isolated from methanol extracts of pear (Pyrus pyrifolia N. cv. Chuhwangbae) fruit peels using solvent fractionation, various types of column chromatogrphy (Diaion HP-20, Sephadex LH-20, and silica gel), and high performance liquid chromatography with an assay guided by 1,1-diphenyl-2-picrylhydrazyl radical-scavenging activity. The isolated compounds were identified as 2-carboxyl-4(1H)-quinolinone (kynurenic acid, 1) from butanol fraction, cis-p-coumaric acid (2) from ethyl acetate-acidic fraction, and vanillin (3) from the ethyl acetate-phenolic fraction, respectively. These isolated compounds were confirmed on the basis of the spectroscopic data of electrospray ionization mass spectrometry and nuclear magnetic resonance. This is the first time that compounds 1-3 were isolated and identified in pear.

Purification and Identification of Apoptosis Modulator Pipernonaline from Piper longum Linn. against Prostate Cancer Cells (필발(Piper longum Linn.)로부터 전립선암 세포사멸물질 pipernonaline의 분리 및 동정)

  • Kim, Kwang-Youn;Kim, Yun-Jin;Lee, Wan;Yu, Sun-Nyoung;Cho, Hyo-Jin;Lee, Sun-Yi;Lee, Han-Seung;Sohn, Jae-Hak;Oh, Hyuncheol;Ahn, Soon-Cheol
    • Journal of Life Science
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    • v.19 no.5
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    • pp.671-675
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    • 2009
  • Prostate cancer has been a critical health problem due to an increase of prostate cancer-related deaths worldwide. Also, a frequent treatment option for prostate cancer is androgen ablation, but this treatment has a limited scope, especially for hormone-refractory cancer. There is an urgent need for the identification of alternative therapeutic strategies for prostate cancer. Previously, over one hundred species of dried-plant methanol extracts were tested for inhibitory effects on proliferation. One of them, Piper longum Linn. was selected based on its potent anti-proliferation effect. The dried root of P. longum Linn. was extracted with 100% methanol for 2-3 days and its extract was fractionated using chloroform. The chloroform layer was then subjected to column chromatography on silica gel, reverse phase-18 (RP-18) and Sephadex LH-20, in turn. Finally, the pure compound was obtained and identified as pipernonaline by NMR spectroscopic and physico-chemical analysis. In this study, anti-proliferation and cell cycle arrest effects of pipernonaline on human prostate cancer PC-3 cells were investigated using the MTT and PI staining, respectively. Our findings suggest that pipernonaline represents a dose-dependent growth inhibition pattern on PC-3 cells and, moreover, its growth inhibition is associated with sub-G1 and G0/G1 cell cycle accumulation in PC-3 cells. Also, these results provide an anticancer candidate for human prostate cancer.

Isolation and Identification of Adenosine and Phlomuroside from the Aerial Parts of Oryza sativa L. (벼(Oryza sativa L.)의 지상부로부터 adenosine과 phlomuroside의 분리 및 동정)

  • Jeong, Rak-Hun;Lee, Dae-Young;Cho, Jin-Gyeong;Baek, Yoon-Su;Seo, Kyeong-Hwa;Lee, Dong-Geol;Kang, Hee-Cheol;Kim, Ji-Young;Baek, Nam-In
    • Journal of Applied Biological Chemistry
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    • v.57 no.4
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    • pp.321-324
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    • 2014
  • Fresh and chopped aerial parts of Oryza sativa were extracted in 80% aqueous mehthanol, and the concentrated extract was successively partitioned in n-hexane, ethyl acetate (EtOAc), n-butanol (n-BuOH), and $H_2O$ fractions. From the n-BuOH fraction, two compounds were isolated through repeated silica gel and ODS column chromatography (c.c.). Based on nuclear magnetic resonance (NMR), mass spectrometry and infrared spectroscopy spectroscopic data, the compounds were identified to be adenosine (1) and phlomuroside (2). Especially, the configuration of both the anomer hydroxyl groups was determined as ${\beta}$ from the coupling constants of the anomer protons (J =6.0 and 7.6 Hz) in the $^1H-NMR$ spectra. This is the first report for the isolation of these compounds from Oryza sativa L.

Inhibition of Low Density Lipoprotein-oxidation, ACAT-1, and ACAT-2 by Lignans from the Bark of Machilus thunbergii

  • Shrestha, Sabina;Park, Ji-Hae;Lee, Dae-Young;Cho, Jin-Gyeong;Lee, Do-Gyeong;Cho, Moon-Hee;Jeong, Tae-Sook;Kang, Hee-Cheol;Baek, Nam-In
    • Journal of Applied Biological Chemistry
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    • v.54 no.1
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    • pp.63-66
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    • 2011
  • The bark of Machilus thunbergii was extracted with 80% aqueous methanol (MeOH), and the concentrated extract was partitioned using ethyl acetate (EtOAc), butanol (n-BuOH), and $H_2O$, successively. From the EtOAc fraction, five lignans were isolated through the repeated silica gel, octadecyl silica gel (ODS) and, Sephadex LH-20 column chromatography. Based on nuclear magnetic resonance (NMR), mass spectroscopy (MS), and infrared spectroscopy (IR) spectroscopic data, the chemical structures of the compounds were determined to be machilin A (1), machilin F (2), licarin A (3), nectandrin A (4), and nectandrin B, (5). This study presents comparative account of five lignans from M. thunbergii bark contributing inhibition of low density lipoprotein (LDL), ACAT-1, and ACAT-2. Compounds 2-5 showed varied degree of antioxidant activity on LDL with $IC_{50}$ values of 2.1, 11.8, 15.3, and $4.1{\mu}M$. Compounds 1, 2, and 3 showed inhibition activity on ACAT-1 with values $63.4{\pm}6.9%$ ($IC_{50}=66.8{\mu}M$), $53.7{\pm}0.9%$ ($IC_{50}=109.2{\mu}M$), and $78.7{\pm}0.2%$ ($IC_{50}=40.6{\mu}M$), respectively, at a concentration of 50 mg/mL, and on ACAT-2 with values $47.3{\pm}1.5%$ ($IC_{50}=149.7{\mu}M$), $39.2{\pm}0.2%$ ($IC_{50}=165.2{\mu}M$), and $52.1{\pm}1.0%$ ($IC_{50}=131.0{\mu}M$, respectively, at a concentration of 50 mg/mL.

Induction of Apoptotic Cell Death in Human Jurkat T Cells by a Chlorophyll Derivative (Cp-D) Isolated from Actinidia arguta Planchon

  • Park, Youn-Hee;Chun, En-Mi;Bae, Myung-Ae;Seu, Young-Bae;Song, Kyung-Sik;Kim, Young-Ho
    • Journal of Microbiology and Biotechnology
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    • v.10 no.1
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    • pp.27-34
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    • 2000
  • The chloroform and methanol (2;1, v/v) extract from an edible plant, Actinidia arguta Planchon, appeared to possess antitumor activity against human leukemias Jurkat T and U937 cells through inducing apoptosis. The substance in the solvent extract was purified by silica gel column chromatography, preparative TLC, and Sephadex LH-20 column chromatography. Characteristics of the substance analyzed by UV scanning analysis, $^1H$ and $^{13}C$ NMR spectra suggested that the substance belongs to the chlorophyll derivatives-like group. The $IC_{50}$ value of the chlorophyll derivative (Cp-D) determined by MTT assay was $15\mu\textrm{g}/ml$ for Jurkat, $10\mu\textrm{g}/ml$ for U937, and $11.4\mu\textrm{g}/ml$ for HL-60m and was more toxic to these leukemias than to solid tumors or normal fibroblast. In order to elucidate cellular mechanisms underlying the cytotoxicity, the effect of the Cp-D on Jurkat T cells was investigated. When cells were treated with the Cp-D at a concentration of $15\mu\textrm{g}/ml$, [3H]thymidine incorporation declined rapidly and wa undetectable in 1h. However, no significant changes were made in the cell cycle distribution of the cells by 24h. The sub-Gl peak representing apoptotic cells began to be detectable in 36h, at which time apoptotic DNA fragmentation was also detected on agarose gel electrophoresis, demonstrating that the cytotoxic effect of the Cp-D is attributable to the induced apoptosis. Under the same conditions, although the protein level of cyclin-dependent kinases such as cdc4, csk6, cdk2, and cdc2 was not significantly changed until 24h, the kinase activity of all c안 rapidly declined and reached a minimum level within 1-6h and then recovered to the initial level by 12h and sustained until 24h. These results suggest that inactivation of cdks at an inappropriate time during the cell cycle progression in jurkat T cells following a treatment with the Cp-D leads to induction of apoptotic cell death.

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Catechin Content and Antioxidative Effect from Rosa davurica Pall (생열귀나무의 카테킨 함량 및 항산화효과)

  • Shin, In-Cheol;Jeong, Kyung-Jin;Shim, Tae-Heum;Oh, Heung-Seok;Park, Sang-Kyun;Cheung, Eui-Ho;Kim, Suk-Nam;Kim, Gwang-Gee;Choi, Dae-Sung;Kwon, Yong-Soo;Kim, Chang-Min;Sa, Jae-Hoon
    • Korean Journal of Pharmacognosy
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    • v.33 no.3 s.130
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    • pp.177-181
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    • 2002
  • This study was carried out to investigate the antioxidative activities of Rosa davurica Pall for the purpose of development of novel antioxidant from natural products. Antioxidant activities of four different parts of Rosa davurica Pall such as fruit, leaf, stem and root were examined by measuring the radical scavenging effect on 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical. The methanol extract from the root of Rosa davurica Pall showed the highest antioxidative activity among 16 samples tested. And, we also tested radical scavenging effects of 5 different extract compartments(Hexane, $CHCl_3$, EtOAc, BuOH and $H_2O$ fraction). EtOAc and BuOH fractions from the root of Rosa davurica Pall exhibited antioxidative activities higher to those of natural, ${\alpha}-tocopherol$ or synthetic antioxidants, BHT. The antioxidative substance of EtOAc fraction from the root of Rosa davurica Pall was successively purified with silica gel adsorption column chromatography and Sephadex LH-20 column chromatography. The purified active substance was isolated as crystal and identified as (+)-catechin by $^{l}H-NMR$ and $^{13}C-NMR$. This compound exhibited DPPH radical scavenging activity with the $IC_50$ value of $1.7\;{\mu}g/ml$. In the analysis of catechin content, the leaf extracts contained the highest catechin, and fruit extracts contained the lowest catechin. Considering antioxidative activity on DPPH assay, the extracts of Rosa davurica Pall showed a possibility to be used as a new material for natural antioxidant and functional food.

Anti-wrinkle Compounds Isolated from the Seeds of Arctium lappa L. (우방자에서 분리한 주름개선 화합물)

  • Hwang, Ju-Young;Park, Tae-Soon;Kim, Dong-Hee;Hwang, Eun-Young;Lee, Jung-Noh;Lee, Ji-Young;Lee, Ghang-Tai;Lee, Kun-Kook;Son, Jun-Ho
    • Journal of Life Science
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    • v.22 no.8
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    • pp.1092-1098
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    • 2012
  • This study was carried out to discover the skin wrinkle reducing components in the seeds of Arctium lappa. The isolation of a methylene chloride-soluble fraction of 70% ethanol extract from the seeds of Arctium lappa using a procollagen type-1 synthesis and MMP-1 activity resulted in the isolation and identification of four lignin compounds: arctiin, arctigenin, matairesinol, and diarctigenin. All structures were confirmed via NMR and MS spectroscopic data. To determine cell viability and procollagen type-1 synthesis, human dermal fibroblasts were treated with 10-100 ${\mu}M$. As a result, none of the four compounds showed cytotoxicity up to 50 ${\mu}M$. We also investigated their procollagen type-1 synthesis and MMP-1 inhibition activity and found that arctiin had the highest activity in terms of both procollagen synthesis and MMP-1 inhibition among all four compounds. Putting all the data together, we suggest that arctiin be used in cosmetics as an anti-wrinkle material.

Isolation and Identification of Antimicrobial Compound from Amarantus lividus (참비름 추출물에서 항균성 물질의 분리 및 동정)

  • Oh, Young-Sook;Lee, Shin-Ho
    • Microbiology and Biotechnology Letters
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    • v.33 no.2
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    • pp.123-129
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    • 2005
  • Isolation and identification of pathogens from slaughter and meat processing plant were investigated. Antimicrobial activity of Amaranthus lividus against isolated pathogens such as Aeromonas sobria, Escherichia coli, Escherichia coli O157, Listeria monocytogenes, Salmonella spp., and Staphylococcus aureus was investigated. Among the chloroform, ethyl acetate and buthanol fraction of amaranthus lividus showed inhibitory effect against Aeromonas sobria CLFM1 and Escherichia coli CLFM2. Antimicrobial substance in chloroform fraction was isolated by silica gel adsorption column chromatography, sephadex LH-20 column chromatography and silica gel partition column chromatography. The antimicrobial compound of amaranthus lividus was identified as diethyl phtalate by HPLC, GC-MS, H-NMR and C-NMR.

Antibacterial Activities of Caesalpinia sappan L. Extract and Structural Analysis of Its Related Brazilin (소목 추출물의 항균활성과 Brazilin의 구조분석)

  • Kwon, Hyun-Jung;Kim, Yong-Hyun;Nam, Kung-Woo;Kim, Sun-Ki;Bang, In-Soek;Han, Man-Deuk
    • Microbiology and Biotechnology Letters
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    • v.38 no.1
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    • pp.105-111
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    • 2010
  • Caesalpinia sappan L. has long been commonly used in oriental folk medicines to treat diseases. To investigate the antibacterial effects from C. sappan L. heart wood, the MeOH soluble extract was successively fractionated by using hexane, $CHC1_3$, EtOAc, BuOH, MeOH, and $H_2O$. Among of these extracts, the EtOAc fraction which partitioned to 3.94% of the highest yields was to be the most active against all human pathogenic bacteria in this experiment. In addition, the antibacterial activities of the EtOAc fraction were more effective against Gram (+) bacteria compared to those against Gram (-) bacteria, which showed difference of the antibacterial activities against Gram (-) bacteria. To confirm the identity of the active substances, the EtOAc fraction was further separated by silica gel adsorption column, high performance liquid chromatography, and 98.48% purity of brazilin (1.67 mg)/EtOAc (10 mg) fraction was obtained from 300 g of C. sappan L. heart wood. The isolated active substance was a single compound of yellow crystalline, and was identified as brazilin ($C_{16}H_{14}O_5$) by MS, and $^lH$-NMR and $^{13}C$-NMR. These results suggest that the brazilin in the EtOAc fraction from MeOH extract of C. sappan L. has a potential as a natural therapeutic agent against human pathogenic Gram (+) bacteria such as Staphylococcus aureus.