• Title/Summary/Keyword: EtOAc fraction

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Anticancer and Signaling Mechanisms of Biologically Active Substances from Orostachys japonicus through Arrest of Cell cycle in Human Melanoma Cells (인체 흑색종 세포에 대한 와송 추출물의 세포주기 억제를 통한 항암효과와 기전 연구)

  • Ryu, Deok-Hyun;Ryu, Deok-Seon
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.32 no.4
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    • pp.1-12
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    • 2019
  • Objectives : The purpose of this study was to identify the anticancer effect of biological substances of ethylacetate(EtOAc) fraction from Orostachys japonicus(OJEF), their effect on human melanoma A375 cells and the related molecular mechanisms. Methods : The MTS assay was used to confirm the inhibition of cancer cell proliferation in A375 cells. And the $MUSE^{TM}$ analyzer was used to determine the ability of OJEF to induce cell cycle arrest. Western blotting was used to determine the changes in protein expression in A375 cells after treatment with OJEF. Results : OJEF showed cytotoxicity to A375 cells. And cell cycle arrest occurred in G1 phase and G2/M phase owing to inhibition of CDK1, cyclin B1, CDK4, and cyclin D, which are related to cell cycle regulation and cell division control. Conclusion : OJEF is effective in regulating cell cycle of human melanoma cells and thus can be a good theraputic agent to treat patients with melanoma.

Hepatoprotective Constituents of the Edible Brown Alga Ecklonia stolonifera on Tacrine-induced Cytotoxicity in Hep G2 Cells

  • Kim, Youn-Chul;An, Ren-Bo;Yoon, Na-Young;Nam, Taek-Jeong;Choi, Jae-Sue
    • Archives of Pharmacal Research
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    • v.28 no.12
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    • pp.1376-1380
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    • 2005
  • In this study, ethanolic extracts from 18 seaweed variants were assessed for hepatoprotective activity against tacrine-induced cytotoxicity in Hep G2 cells. Only one of these, Ecklonia stolonifera Okamura (Laminariaceae), a member of the brown algae, exhibited promising hepatoprotective activity. Bioassay-guided fractionation of the active ethyl acetate (EtOAc) soluble fraction obtained from the ethanolic extract of E. stolonifera, resulted in the isolation of several phlorotannins [phloroglucinol (1), eckstolonol (2), eckol (3), phlorofucofuroeckol A (4), and dieckol (5)]. Compounds 2 and 4 were determined to protect Hep G2 cells against the cytotoxic effects of tacrine, with $EC_{50}$ values of 62.0 and 79.2 $\mu$g/mL, respectively. Silybin, a well characterized hepatoprotective agent, was used as a positive control, and exhibited an $EC_{50}$ value of 50.0 $\mu$g/mL. It has been suggested that the phlorotannins derived from marine brown algae might prove useful sources in the development of novel hepatoprotective agents.

Determination and Isolation of leaf Isoflavone in Hyacinth Bean (제비콩 잎의 isoflavone 함량 및 성분 분석)

  • Kim, Yong-Wook;Lim, Se-Jin;Kim, Myoung-Ae;Choi, Woo-Chul;Yoon, Hong-Tae
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.46 no.6
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    • pp.449-452
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    • 2001
  • Legume seed isoflavones may have a variety of desirable physiological effect on the human health including both the circulatory and skeletal systems. The present study was performed to determine the isoflavone content of leaf and seed as well as to purify and identify the types of isoflavone from leaf extract of hyacinth bean (Lablab purpureus (L.) Sweet). Reverse phase HPLC revealed six different types of isoflavone such as daidzin, genistin, daidzein, genistein, 6"-o-acetyl genistin and 6"-o-acetyl daidzin in aqueous methanol extract from seeds and leaves of the hyacinth bean. Relatively, leaf isoflavone content of hyacinth bean was greater than seed isoflavone content. Using DiAion HP-20 silica gel and sephadex LH-20 chromatography, pure daidzein was identified in the ether layer, whereas genistin was in the EtOAC fraction.

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Antioxidant Constituents of Athyrium acutipinnulum (섬고사리의 항산화 성분)

  • Hye Jin Park;Se Hwan Ryu;Sang Won Yeon;Ayman Turk;Solip Lee;Hak Hyun Lee;Bang Yeon Hwang;Mi Kyeong Lee
    • Korean Journal of Pharmacognosy
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    • v.54 no.2
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    • pp.53-60
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    • 2023
  • Athyrium acutipinnulum, called as Ulleungdo ladyfern (Seom-go-sa-ri), is a native plant of South Korea. A. acutipinnulum has been consumed as foods and also traditionally used for the treatment of epilepsy, gonorrhea and nerve disorder. The methanolic extract and EtOAc soluble fraction of A. acutipinnulum showed the antioxidant activity. Fractionation using various chromatographic techniques resulted in the isolation of 13 compounds. The structures were elucidated on the basis of spectroscopic methods as seven phenolic compounds, methyl 2-hydroxy-3-phenylpropanoate (1), protocatechualdehyde (2), caffeic acid (3), trans-p-coumaric acid (4), (-)-4-E-caffeoyl-L-threonic acid (5), 5-O-caffeoyl shikimic acid (6) and 5-O-caffeoyl quinic acid (7), three flavonoids, quercetin 3-O-β-glucoside (8), naringenin-7-O-β-glucoside (9) and sutchenoside A (10), two steroids, ponasterone A (11) and ecdysone (12) and a coumarin, esculetin (13). Among them, compounds 5 and 10 were first reported from Athyrium spp and compounds 2, 5, 6 and 7 showed the antioxidant activity.

The Lipid Efflux Effects of Dichloromethane Extract from Orostachys japonicus in 3T3-L1 Adipocyte Cells (3T3-L1 지방세포에 대한 와송 디클로로메탄 추출물의 지질 대사 개선에 관한 연구)

  • Kim, Soo-Hwan;Lee, Hyeong-Seon
    • Korean Journal of Clinical Laboratory Science
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    • v.51 no.4
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    • pp.514-520
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    • 2019
  • This study investigated the improved lipid metabolism effect of 3T3-L1 cells induced by adipocytes using the dichloromethane (DCM) fraction in the organic solvent extract of Wassong (Orostachys japonicus). To confirm the cell cytotoxicity, each of 6 fractions of organic solvent extracts (EtOH, Hexane, DCM, EtOAc, BuOH, and H2O) was examined using MTS assay. As a result, it was confirmed that the DCM extract was stable over the whole range of concentrations, and a DCM fraction was used to confirm the improved lipid metabolism effect. Lipid excretion was measured to confirm the change of lipid metabolism. 3T3-L1 cells induced by adipocytes were treated with DCM extract and stained with oil-red O to evaluate lipid accumulation. As a result, it was confirmed that the lipid efflux was significantly improved. In order to confirm the mechanism of lipid efflux, the mRNA expressions of ABCA1 and ABCG1, which are lipid transport proteins, were confirmed by real-time PCR. Therefore, the present study confirmed that the DCM extract from Orostachys japonicus has the effect of improving the lipid metabolism on 3T3-L1 adipocytes. In addition, the results of this study will be used as the basis for the development of functional foods using Orostachys japonicus and also for conducting research on the detailed mechanisms.

Antioxidant Activities of Solvent Extracts from Pomegranate Endocarp (석류 내피 용매별 추출물의 항산화 활성)

  • Jin, So-Yeon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.12
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    • pp.1635-1641
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    • 2011
  • This study was designed to investigate antioxidant activities of pomegranate endocarp extracts. Pomegranate endocarp extract contains the highest antioxidant function compared to pomegranate extracts from other parts. Pomegranate endocarp extract was fractionated with hexane, dichloromethane, ethyl acetate, butanol extract, and water, followed by evaluation for antioxidant activity. During this experiment, various antioxidant tests such as nitrite scavenging activity, reducing power, superoxide anion scavenging activity, $ABTS^+$ scavenging activity, SOD like-activity, and DPPH radical scavenging activity were conducted on the $CH_2Cl_2$, EtOAc, BuOH, $H_2O$ fractions of pomegranate endocarp extract. Results showed that the ethyl acetate fraction contained the highest DPPH radical scavenging activity among the samples. For SOD like-activity, the dichloromethane fraction had the highest antioxidant activity. For superoxide anion scavenging activity, the ethyl acetate fraction had the highest antioxidant activity. In the $ABTS^+$ scavenging activity test, the ethyl acetate fraction $IC_{50}$ was 39.26 ${\mu}g$/mL, whereas that of the butanol fraction was 40.95 ${\mu}g$/mL. In testing reduction power at 0.1 mg/mL, the O.D. of the ethyl acetate fraction was highest at 1.404 and showed higher activity than ascorbic acid at 1.332. The results of the nitrite scavenging activity test were very similar to those of the SOD like-activity test. These results suggest that pomegranate endocarp extract may have value as a natural antioxidant.

Cytotoxic Effect of Flavonoids from the Roots of Glycyrrhiza uralensis on Human Cancer Cell Lines (감초(Glycyrrhiza uralensis Fisch.)로부터 분리된 flavonoid의 인체 암세포에 대한 세포독성)

  • Park, Ji-Hae;Wu, Qian;Yoo, Ki-Hyun;Yong, Hye-Im;Cho, Sueng-Mock;Chung, In-Sik;Baek, Nam-In
    • Journal of Applied Biological Chemistry
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    • v.54 no.1
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    • pp.67-70
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    • 2011
  • The roots of Glycyrrhiza uralensis Fisch. were extracted with 30% aqueous ethanol (EtOH), and the concentrated extract was partitioned with n-hexane, chloroform ($CHCl_3$), ethyl acetate (EtOAc), n-butanol (n-BuOH), and $H_2O$, successively. From the $CHCl_3$ fraction, four flavonoids were isolated through the repeated silica gel ($SiO_2$), octadecyl silica gel (ODS), and Sephadex LH-20 column chromatographies (c.c.). According to the results of spectroscopic data including nuclear magnetic resonance spectrometry (NMR), electron ionization mass spectrometry (EI/MS), and infrared spectroscopy (IR), the chemical structures of the compounds were determined as glabrol (1), abyssinone II (2), glabridin (3), and isoliquiritigenin (4). The flavonoids were evaluated for cytotoxic effect against human cancer cell lines, HCT-116, HepG2, HeLa, SK-OV-3, SK-BR-3, MCF-7, and SK-MEL-5. Especially, glabrol (1) and glabridin (2) showed $IC_{50}$ values of lower than $25{\mu}M$.

Triterpenoids from the Flower of Campsis grandiflora K. Schum. As Human Acyl-CoA: Cholesterol Acyltransferase Inhibitors

  • Kim, Dong-Hyun;Han, Kyung-Min;Chung, In-Sik;Kim, Dae-Keun;Kim, Sung-Hoon;Kwon, Byoung-Mog;Jeong, Tae-Sook;Park, Mi-Hyun;Ahn, Eun-Mi;Baek, Nam-In
    • Archives of Pharmacal Research
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    • v.28 no.5
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    • pp.550-556
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    • 2005
  • The flower of Campsis grandiflora K. Schum. Was extracted with 80% aqueous MeOH, and the concentrated extract was partitioned with EtOAc, n-BuOH and H$_2$O. From the EtO Ac fraction, seven triterpenoids were isolated through the repeated silica gel, ODS column chromatographies and preparative HPLC. From the result of physico- chemical data including NMR, MS and IR, the chemical structures of the compounds were determined as 3${\beta}$-hydroxyolean-12-en-28-oic acid (oleanolic acid, 1), 3${\beta}$-hydroxyurs-12-en-28-oic acid (ursolic acid, 2), 3${\beta}$-hydroxyurs-12-en-28-al (ursolic aldehyde, 3), 2${\alpha}$,3${\beta}$-dihydroxyolean-12-en-28-oic acid (maslinic acid, 4), 2${\alpha}$,3${\beta}$-dihydroxyurs-12-en-28-oic acid (corosolic acid, 5), 3${\beta}$,23-dihydroxyurs-12- en-28-oic acid (23-hydroxyursolic acid ,6) and 2${\alpha}$,3${\beta}$,23-trihydroxyolean-12-en-28- oic acid (arjunolic acid, 7). These teriterpenoids were isolated for the first time from this plant. Also, compounds 4, 5, 6, and 7 revealed relatively high hACAT-1 inhibitory activity with the value of 46.2${\pm}$1.1, 46.7${\pm}$0.9, 41.5${\pm}$1.3 and 60.8${\pm}$1.1% at the concentration of 100${\mu}$g/mL, respectively.

Anti-Inflammatory Effect of Flavonoids from Brugmansia arborea L. Flowers

  • Kim, Hyoung-Geun;Jang, Davin;Jung, Young Sung;Oh, Hyun-Ji;Oh, Seon Min;Lee, Yeong-Geun;Kang, Se Chan;Kim, Dae-Ok;Lee, Dae Young;Baek, Nam-In
    • Journal of Microbiology and Biotechnology
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    • v.30 no.2
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    • pp.163-171
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    • 2020
  • Brugmansia arborea L. (Solanaceae), commonly known as "angel's trumpet," is widely grown in North America, Africa, Australia, and Asia. It has been mainly used for ornamental purposes as well as analgesic, anti-rheumatic, vulnerary, decongestant, and anti-spasmodic materials. B. arborea is also reported to show anti-cholinergic activity, for which many alkaloids were reported to be principally responsible. However, to the best of our knowledge, a phytochemical study of B. arborea flowers has not yet been performed. Four flavonol glycosides (1-4) and one dihydroflavanol (5) were for the first time isolated from B. arborea flowers in this study. The flavonoids showed significant antioxidant capacities, suppressed nitric oxide production in lipopolysaccharide (LPS)-treated RAW 264.7 cells, and reduced inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX-2) protein production increased by LPS treatment. The contents of compounds 1-4 in n-BuOH fraction were determined to be 3.8 ± 0.9%, 2.2 ± 0.5%, 20.3 ± 1.1%, and 2.3 ± 0.4%, respectively, and that of compound 5 in EtOAc fraction was determined to be 12.7 ± 0.7%, by HPLC experiment. These results suggest that flavonol glycosides (1-4) and dihydroflavanol (5) can serve as index components of B. arborea flowers in standardizing anti-inflammatory materials.

Antiamoebic Activity of Petiveria alliacea Leaves and Their Main Component, Isoarborinol

  • Zavala-Ocampo, Lizeth M.;Aguirre-Hernandez, Eva;Perez-Hernandez, Nury;Rivera, Gildardo;Marchat, Laurence A.;Ramirez-Moreno, Esther
    • Journal of Microbiology and Biotechnology
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    • v.27 no.8
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    • pp.1401-1408
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    • 2017
  • Petiveria alliacea L. (Phytolacaceae) is a medicinal plant with a broad range of traditional therapeutic properties, including the treatment of dysentery and intestinal infections caused by protozoan parasites. However, its effects against Entamoeba histolytica have not been reported yet. We investigated the antiamoebic activity present in the leaves of P. alliacea Antiamoebic activity was evaluated in methanolic and aqueous extracts, as well as in the hexanic, methanolic, and EtOAc fractions. The P. alliacea methanolic extract showed a better antiamoebic activity than the aqueous extract with an $IC_{50}=0.51mg/ml$. Likewise, the hexanic fraction was the most effective fraction, showing a dose-dependent activity against E. histolytica, with an $IC_{50}=0.68mg/ml$. Hexanic subfraction 12-19 showed the highest antiamoebic activity at 0.8 mg/ml, producing 74.3% growth inhibition without any toxicity in mammal cells. A major component in subfraction 12-19 was identified as isoarborinol, which produced 51.4% E. histolytica growth inhibition at 0.05 mg/ml without affecting mammal cells. The P. alliacea leaf extract has antiamoebic activity that can be attributed to a major metabolite known as isoarborinol.