Inhibitory Effects of Flavonoids Isolated from the Leaves of Stewartia koreana on Nitric-oxide Production in LPS-stimulated RAW 264.7 Cells

Seung-Su Lee(이승수),Myun-Ho Bang(방면호),Se-Ho Park(박세호),Dae-kyun Chung(정대균),Seun-Ah Yang(양선아) 한국생명과학회 2018 생명과학회지 Vol.28 No.5

노각나무(Stewartia koreana) 잎 에틸아세테이트 분획으로부터 quercetin (1), quercitrin (2), hyperin (3), quercetin-3-O-(6""-O-galloyl)-β-D-galactopyranoside (4), kaempferol-3-o-[2"",6""-di-o-(trans-p-coumaroyl)]-β-D-glucopyranoside (5)의 5종의 플라보노이드를 분리하였으며, 이들 5종 성분의 염증 반응에 대한 활성을 분석하기 위하여 LPS를 처리한 대식세포에서 산화질소(NO) 생성 억제활성을 측정하였다. 이들 5종 성분 중 compound 4, 5는 노각나무에서 처음으로 분리된 것으로 항염증 활성에 대한 보고도 없다. 분광분석법으로 확인된 노각나무 잎 유래성분들은 LPS 처리한 대식세포의 NO 생성을 유의적으로 저해하였으며, 특히 kaempferol-3-o-[2"",6""-di-o-(transp-coumaroyl)]-β-D-glucopyranoside (5)는 가장 강한 억제효과(17.17%, 5.0%, 3.92%, 6.32% and 63.35% inhibition of compound 1, 2, 3, 4 and 5 at 10 μg/ml)를 나타냈다. 또한, 이러한 NO 생성 억제효과는 inducible nitric oxide synthase(iNOS) 단백질 발현 억제를 통한 것으로 나타났다. 따라서, 본 연구에서 새로 분리된 플라보놀인 kaempferol-3-o-[2"",6""-di-o-(trans-p-coumaroyl)]-β-D-glucopyranoside (5)는 노각나무 잎의 항염증 활성을 나타내는 주요 물질로 사료된다. Five phenolic compounds were isolated from the ethyl acetate fraction of leaves from Stewartia koreana, and their nitric-oxide (NO) inhibitory activities were measured to identify the major active constituents responsible for the efficacy of the extract against inflammatory reactions. These five compounds were quercetin (1), quercitrin (2), hyperin (3), quercetin-3-O-(6""-O-galloyl)-β-D-galactopyranoside (4), and kaempferol 3-O-[2"",6""-di-O-(trans-p-coumaroyl)]-β-D-glucopyranoside (5). Among the separated compounds in the EtOAc fraction, compounds 4 and 5 were isolated for the first time, and no study has yet reported their anti-inflammatory effects. The compounds were identified by spectroscopic analysis, and the isolated compounds showed significant NO inhibitory effects in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. Compound 5 showed the most potent inhibitory effect (63.35% inhibition) against LPS-induced NO production compared to that of compound 1 (17.17%), compound 2 (5.0%), compound 3 (3.92%), and compound 4 (6.32%) at 10 μg/ml concentration. NO production was inhibited by suppressing the protein expression of inducible nitric-oxide synthase in LPS-stimulated RAW 264.7 macrophages. These results indicate that kaempferol 3-O-[2"",6""-di-O-(trans-p-coumaroyl)]-β-D-glucopyranoside might be the major active compound responsible for the anti-inflammatory effects of S. koreana.

동부로부터 sterol의 분리 동정

최은희,박희정,吴倩,정인식,김지영,백남인 한국응용생명화학회 2010 Journal of Applied Biological Chemistry (J. Appl. Vol.53 No.2

The seed of cowpea (Vigna sinensis K.) was extracted with 80% aqueous methanol (MeOH). And the concentrated extract was partitioned with ethyl acetate (EtOAc), n-butanol (n-BuOH) and H2O, successively. The repeated silica gel and octadecyl silica gel (ODS) column chromatographic separations for the EtOAc and n-BuOH fractions led to isolation of four sterols. And the chemical structures of the compounds were determined as a mixture of stigmasterol and β-sitosterol with the ratio of 4 to 3 (1), 7-ketositosterol (2), and stigmasterol 3-O-β-D-glucopyranoside (3) from the interpretation of spectroscopic data including nuclear magnetic resonance (NMR) spectrum metric, mass (MS) spectrum metric and infrared (IR) spectroscope. This study reports the first isolation of β-sitosterol, 7-ketositosterol, and stigmasterol 3-O-β-D-glucopyranoside from the seed of Vigna sinensis K. In addition, compound 2, 7-ketositosterol, is rarely occurred in natural source including plant. 동부를 80% MeOH로 추출하고, 얻어진 추출물을 EtOAc, n-BuOH 및 H2O로 용매 분획 하였다. 이 중 EtOAc과 n-BuOH 분획으로부터 silica gel과 octadecyl silica gel(ODS) columnchromatography를 반복하여 4종의 sterol 화합물을 분리, 정제하였다. 각 화합물의 화학구조는 NMR, MS 및 IR 등의 spectrum data를 해석하여, stigmasterol(1a), β-sitosterol(1b), 7-ketositosterol(2) 및 stigmasterol 3-O-β-D-glucopyranoside(3)로 동정하였다. 화합물 1b, 2와 3은 동부에서 처음 분리되었다.

Evaluation of Salicornia herbacea as a Potential Antioxidant and Anti-Inflammatory Agent

You Ah Kim,Chang-Suk Kong,Young Ran Um,Sung Su Yea,서영완 한국식품영양과학회 2009 Journal of medicinal food Vol.12 No.3

In this study, the antioxidant and anti-inflammatory activities of Salicornia herbacea were evaluated. The crude CH2Cl2/methanol extract of S. herbacea showed 52% and 86% scavenging activities of the authentic ONOO− and ONOO− from 3-morpholinosydnomimine (SIN-1) at a concentration of 50μg/mL, respectively, and was subjected to a further fractionation with n-hexane, 85% aqueous methanol, n-butanol, and water. Additional purification of the n-butanol fraction revealed that the most potent scavenging activity led to the isolation of isorhamnetin 3-O-β-d-glucopyranoside as the active principle. The structure of isorhamnetin 3-O-β-d-glucopyranoside was elucidated by extensive two-dimensional nuclear magnetic resonance experiments such as 1H correlation spectroscopy nuclear Overhauser effect spectroscopy, heteronuclear single quantum correlation, and heteronuclear multiple-bond correlation as well as by comparison with the published spectral data. Isorhamnetin 3-O-β-d-glucopyranoside exhibited dose-dependent scavenging activities of the authentic ONOO− and ONOO− from SIN-1. The electron spin resonance spin-trap techniques confirmed that reactive oxygen species, including the hydroxyl, superoxide, carbon-centered, and 1,1-diphenyl-2-picrylhydrazyl radicals, were actively quenched by addition of isorhamnetin 3-O-β-d-glucopyranoside. In addition, isorhamnetin 3-O-β-d-glucopyranoside suppressed the lipopolysaccharide-induced nitric oxide production and the expression of cytokines such as inducible nitric oxide synthase, tumor necrosis factor-α, and interleukin-1β in Raw 264.7 cells. Findings from this study should underscore the nutraceutical value of S. herbacea-derived isorhamnetin 3-O-β-d-glucopyranoside as a potent antioxidative and anti-inflammatory agent via alleviation of radical-induced toxicities and pro-inflammatory responses.

산오이풀 잎의 성분

권원준(Won Jun Kwon),황완균(Wan Kyunn Whang),김일혁(Il Hyuk Kim) 대한약학회 1996 약학회지 Vol.40 No.3

The constituents of Sanguisorba hakusanensis leaves (Rosaceae), of which the roots have been used as an astringent, hemostatics and antiphlogistics, were studied phytochemically. From water fraction of the MeOH extract, gallic acid 3-O-beta-D-(6''-O-galloyl)-glucopyranoside(I), quercetin-3-O-beta-D-galactopyranoside(II), quercetin-3-O-alpha-L-arabinopyranoside(III) and 2alpha, 3beta, 19alpha, 23-tetrahydroxyurs-12-en-28-oic acid 28-O-beta-D-glucopyranoside(IV) were isolated by column chromatographic separation using Amberlite XAD-2, ODS-gel and Sephadex LH-20. The structure of these compounds were elucidated by spectroscopic parameters of 1H-NMR, 13C-NMR, 1H-1H COSY, 13C-1H COSY, EI-Mass, FAB-Mass, IR, UV and by comparison with authentic samples.

목통의 페놀성 화합물의 항산화 활성 및 HPLC-UV 동시 함량분석

경민지(Min-Ji Kyung),전혜진(Hye-Jin Jeon),황완균(Wan-Kyunn Whang) 大韓藥學會 2015 약학회지 Vol.59 No.3

Akebiae caulis have been used in folk medicines for diuretic, menstrual pain, and diuretic pain. It has been also resolved nephritis and cystitis. In this study, we isolated three phenolic compounds from 70% methanol extract using the open column chromatography. These isolated compounds which were 2-(3,4-dihydroxyphenyl)-ethyl-β-D-glucopyranoside, 5-O-caffeoylquinic acid, and syringoylglycerol 2-O-β-D-glucoside were determined by physico-chemical apparatus. Furthermore, we conducted DPPH and ABTS assay in order to screen the antioxidant activity of isolated three compounds. Also, we developed a rapidly HPLC-UV analysis method of two phenolic compounds (2-(3,4-dihydroxyphenyl)-ethyl-β-Dglucopyranoside, 5-O-caffeoylquinic acid) for evaluating the Akebiae Caulis collected 30 samples from different regions. From the experiments, all three isolated compounds showed the significant antioxidant activity. We suggested that the content criteria of marker compounds were shown by a simple and rapid HPLC-UV method. The contents respectively were 0.009% (2-(3,4-dihydroxyphenyl)-ethyl-β-D-glucopyranoside) and 0.036% (5-O-caffeoylquinic acid).

Flavonoids from the Leaves of Polygala japonica

Do, Jae Chul,Yu, Young Jun,Jung, Keun Young,Son, Kun Ho 영남대학교 약품개발연구소 1992 영남대학교 약품개발연구소 연구업적집 Vol.2 No.-

From the leaves of Polygala japonica, kaempferol (1), astragalin (2), kaempferol 3-O-(6"-O-acetyl)-β-_(D)-glucopyranoside (3) and kaempferol 3, 7-di-O-β-_(D)-glucopyranoside (4), have been isolated and characterized by chemical and spectral means.

Chemical Constituents from Acer mandshuricum and Their Effects on the Function of Osteoblastic MC3T3-E1 Cells

Yan Ding,Chun Liang,Huu Tung Nguyen,Eun Mi Choi,김정아,김영호 대한화학회 2010 Bulletin of the Korean Chemical Society Vol.31 No.4

A new compound, 4-methoxyl 5-hydroxymethyl benzoic 3-O-β-D-glucopyranoside (1), has been isolated from the leaves and stems of Acer mandshuricum, along with nine known compounds (2-10). Their structures were determined by a variety of spectroscopic analyses. The effect of compounds 1-10 on the function of osteoblastic MC3T3-E1 cells was examined by determining alkaline phosphatase (ALP) activity, collagen synthesis, and mineralization. Compound 1significantly increased the function of osteoblastic MC3T3-E1 cells; 5.0 μM of 1 increased ALP activity, collagen synthesis,and mineralization of MC3T3-E1 cells to 114.7, 119.5, and 108.2% (P < 0.05) of the basal value, respectively. In addition, compounds 2-10 also potently increased the function of osteoblastic MC3T3-E1 cells.

Effects of the Methanol Extract of the Leaves of Brassica juncea and Its Major Component, Isorhamnetin 3-o-β-D-Glucoside, on Hepatic Drug Metabolizing Enzymes in Bromobenzene-treated Rats

Jong Moon Hur,Jong Won Choi,Jong Cheol Park 한국식품과학회 2007 Food Science and Biotechnology Vol.16 No.3

The effects of the methanol extract of the leaves of Brassica juncea and isorhamnetin 3-o-β-D-glucopyranoside, major compound isolated from the ethyl acetate fraction of this plant on hepatic lipid peroxidation and drug-metabolizing enzymes, were evaluated in rats treated with bromobenzene. The extract and isorhamnetin 3-o-β-D-glucopyranoside of oral administration did not show any significant effects on activities of aminopyrine N-demethylase and aniline hydroxylase, enzymes forming toxic epoxide by bromobenzene as well as on glutathione content. However, both methanal extract and isorhamnetin 3-o-β-D-glucopyranoside significantly recovered the decreased activities of lutathione s-transferase and epoxide hydrolase, and also reduced the lipid peroxide level in rats treated with bromobenzene. From the results, the protections of this plant against bromobenzene-induced hepatotoxicity are thought to be via enhancing the activities of epoxide hydrolase and glutathione s-transferase, enzymes removing toxic epoxide, and reducing the lipid peroxide level.

연방(蓮房)의 티로시나제 저해 활성을 보이는 성분분리

조현우,정수영,안병관,정원석,조정희 한국생약학회 2013 생약학회지 Vol.44 No.1

Nelnumbo nucifera Gaerth. (Nymphaeaceae) has been used in a korean traditional medicine to treat fever, sunstroke and dizziness. The receptaculums of this plant were refluxed with MeOH, and then fractionated with organic solvents (CH2Cl2,EtOAc and n-BuOH) to screen whitening activity using tyrosinase inhibitory activity. EtOAc (IC50, 45.23 µg/ml) fractions showed a good tyrosinase inhibitory activity. Column chromatographic separation of CH2Cl2 and EtOAc fractions of Receptaculum nelumbinis led to the isolation 3 compounds. Their chemical structures were characterized as β-sitosterol (1), quercetin 3-O-β-D-galactopyranoside (2) and kaempferol 3-O-β-D-glucopyranoside (3) by comparison NMR spectral data and with those in references, respectively. Isolated compounds 1 and 3 were firstly isolated from Receptaculums nelumbinis. Compounds 2and 3 showed potent whitening activities.

홍화의 성분 분리 및 항산화 활성

이승호,최현규,강연복,박성희,손애량,나민균 한국생약학회 2011 생약학회지 Vol.42 No.2

As part of our ongoing study focused on the discovery of antioxidants from natural products by measuring the DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging activity, methanol extract of flowers of Carthamustinctorius L. was found to show potent antioxidant activity. Activity-guided fractionation of the methanol extract lead to the isolation of twenty compounds including two flavonol glycosides, quercertin-3-O-β-D-glucopyranoside (12) and kaempferol 3-O-α-L-rhamnopyranosyl-β-D-glucopyranoside (18), two flavanone glycosides, (2S)-4',5,6,7-tetrahydroxyflavanone 6-O-β-D-glucopyranoside (15) and (2R)-5,7,8',4-tetrahydroxyflavanone 8-O-β-D-glucopyranoside (16), and two acetylenic glycosides, 8Z-decaene-4,6-diyne-1-O-β-D-glucopyranoside (13) and 4,6-decadiyne-1-O-β-D-glucopyranoside (14). Their chemical structures were identified by using spectroscopic analysis. Among them, compounds 12-18 were tested in DPPH assay. Compounds 13-16 were first reported to their antioxidant activity. Quercertin-3-O-β-D-glucopyranoside (12) showed the most potent inhibitory effect on DPPH with IC_50 value of 56.7 μM.