• 대한본초학회지
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• 제25권1호
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• pp.13-21
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• 2010

Objectives : This study was purposed to the anti-oxidative effects of Polygonati Multiflori Caulis(henceforth PMC) on oxidized brain and liver cells in rats. Methods : After extraction of PMC with water, the water extract was divided into five fractions : hexane, ethyl ether, ethyl acetate, butanol and an aqueous fraction. The phenol contents of each fraction were measured. The lipid peroxidation inhibition effect were then investigated. Results : After processing PCM water and PCM fractionations on oxidized brain cells in rats, the SOD (super oxide dismutase) activity and glutathione content were increased, and the NO (nitric oxide) content was decreased. It had much higher SOD activity than liver cells in rats excluded in the n-BuOH and aqueous fractions. In case of oxidized liver cells in rats, the SOD activity and glutathione content increased, while both the NO content and the MDA (malondialdehyde) content decreased. It had much higher glutathione content than brain cells in rats in the every fractions. It had much lower MDA content than brain cells in rats in the Aqueous fractions and brain cells in rats had much lower MDA content than liver cells in rats in the total extract, n-hexane, EtOEt, EtOAc and n-BuOH fractions. Conclusions : PMC has anti-oxidative effect on oxidized liver cells and brain cells in rats, through there are differences in fraction. Additionally, Anti-oxidative effect of brain cells can be relaxed the mental nerve and it is related PMC effect.

• 한국약용작물학회지
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• 제13권6호
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• pp.217-225
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• 2005

강원도 산지의 뽕나무의 잎, 줄기, 뿌리를 건조하여 100% EtOH와 물로 추출하여 얻은 추출물 및 EtOH추출물로부터 용매별 분획물들을 NIH/3T3에 대한 세포독성, comet assay를 이용한 DNA damage손상으로부터 유전 독성, in vitro micronucleus assay를 이용한 유전독성, Ames test를 이용한 돌연변이 유발을 조사하여 다음과 같은 결과를 얻었다. 1) 각 시료들의 NIH/3T3에 대한 세포독성은 0.5mg/ml 농도에서 약 80%의 생육활성을 나타냈다. 2)뽕나무 잎의 1차 추출물들은 comet assay에서 양성 대조구 대비 약 40% 정도의 유전독성을 보였다. 뽕잎의 EtOH 추출물에서 분획한 분획물 시료중 수용성 분획은 거의 독성이 나타나지 않았으나, hexan의 0.25mg/ml 농도의 분획물이 약 50%정도를 보였으며 다른 시료들은 이 보다 약한 독성을 보였다. 또한 뽕나무 줄기의 분획물 시료들 중에서 0.5mg/ml 농도의 chloroform 분획물이 양성 대조구 대비 가장 강한 50%의 유전독성을 보인것 외에 다른 시료들은 그보다 약한 독성을 보였다. 3) In vitro micronucleus assay를 이용한 유전독성은 재료의 1차 추출물은 거의 유전독성이 나타나지 않았고 EtOH 추출물의 재분획물인 시료중에서 0.5mg/ml의 농도의 hexan 및 chloroform 분획물은 MN의 발생 빈도가 음성대조구에 비하여 약 2배의 증가로 나타났다 또 뽕나무 줄기의 분획물 시료들에서 0.5mg/ml의 hekan, chloroform 및 EtOAc 분획물에서 MN이 관찰 되었으나 그중 chloroform 분획물 시료에서 더 많은 MN 발생을 보였다. 4) Ames test를 이용한 돌연변이 실험결과 뽕나무 잎 1차 추출물들은 0.25mg/ml의 농도에서 TA98에 대하여 거의 돌연변이 현상이 없었고, EtOH 추출물로부터 재 분획한 BuOH 및 chloroform분 획물에서는 강한 돌연변이 현상이, 또 5mg/ml의 hekan 분획물에서는 더욱 강한 돌연변이 현상이 관찰되었다. TA100 strain에 대하여는 각 재료의 1차 추출물이나 분획들의 낮은 농도 (1.25mg/ml)에서는 돌연변이 현상이 매우 약하거나 거의 없었고, 0.25mg/ml의 1차 추출물들에서는 매우 약한 돌연변이가 나타났으나 이들의 EtOAc 분획물은 다소 강한 돌연변이 유발성이 관찰 되었다. 그러나 수용성 분획물은 두 균주에 대하여 전혀 돌연변이 유발성을 보이지 않았다. 5) 뽕나무 줄기로부터 조제한 시료중 수용성 분획물과 EtOAc분획물은 IA98에서 전혀 돌연변이 유발성이 없었으나 0.25mg/ml 농도외 물 추출물, EtOH 춘출물, hexan 및 chloroffrm분획물들이 약 40%증가한 복귀변이 콜로니가 관찰되었고, 이보다 농도를 증가시킨 물 추출물 및 BuOH 분획물에서도 같은 수준의 복귀변이 콜로니가 관찰 된 결과로부터 돌연변이 유발성이 있음이 확인되었다 TA100에 대하여 뽕나무 줄기의 수용성 분획물은 전혀 돌연변이가 나타나지 않았고, 0.25mg/ml의 물 추출물, EtOH추출물, hexan 및 BuOH분획물들이 약 10%증가한 약한 돌연변이가 관찰되었다. 그러나 농도를 5 mg/ml로 한 EtOAc 분획물은 26의 돌연변이 유발성을 보였다. 이상의 결과로 부터 뽕나무를 이용한 식용 제품생산을 고려할 때 추출물들의 제조와 선택을 가름하는 자료로서의 활용이 기대되며 앞으로 in vivo test 등 더욱 연구할 필요가 있을 것으로 사료된다.

• 한국식품영양과학회지
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• 제29권5호
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• pp.900-907
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• 2000

The antioxidant effects of dichloromethane, ethylacetate or water fraction of kimchi added to the 1% cholesterol diet were studied. Six New Zealand white rabbits in each group were fed either control diet (basal diet containing 1% cholesterol) or experimental diet containing dichloromethane (CH$_2$Cl$_2$), ethylacetate (EtOAc) or water ($H_2O$) fraction of kimchi in the control diet for 16 weeks. The amount of each solvent fraction of kimchi added to the experimental diet was equivalent to 5% of freeze-dried kimchi. Levels of hepatic lipid oxidation expressed as TBARS or peroxide value for the experimental groups were lower than that of control (p<0.05). Liver homogenated of the experimental group containing dichloromethane fraction of kimchi inhibited LDL oxidation in the presence of Cu++ by 46% (p<0.05). The activities of catalase, Glutathione peroxidase (GSH-Px), Cu, Zn-superoxide (Cu, Zn-SOD) and Mn-superoxide (Mn-SOD) of experimental groups were lower than those of control group. Low enzyme activities observed from the kimchi solvent fraction groups might be due to the level of lipid oxidation progressed less in these groups. The most significant antioxidant effects were observed from dichloromethane fraction of kimchi among the experimental groups. The major fatty acids of hepatic phospholipid of rabbit were C18:2 and C18:0. But the major fatty acid profile was changed into C16:0, C18:0, C18:1, and C18:2 when rabbit was fed 1% cholesterol diet for 16 weeks, and this profile was almost the same as in rabbit fed diet containing kimchi solvent fraction. The ratio for unsaturated fatty acid to saturated fatty acid decreased by cholesterol induced diet and it was not corrected by kimchi solvent fractions.

• Archives of Pharmacal Research
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• 제25권6호
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• pp.865-872
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• 2002

In the course of the investigations of natural antioxidants, we examined the antioxidant activities of the methanol (MeOH) extracts of some selected Prunus species, including P. buergeriana, P. davidiana, P padus, P. pendula for. ascendens, P. sargentii, P. serrulata var. spontanea and P. yedoensis by three methods as represented by the 1, 1-diphenyl-2-picrylhydrazyl (DPPH) radical, total ROS (reactive oxygen species) and the peroxynitrite ($ONOO^-$) scavenging activity tests. We also evaluated the activities of the organic solvent-soluble fractions, including the dichloromethane ($CH_2Cl_2$), ethyl acetate (EtOAc), n-butanol (n-BuOH) fractions and the water ($H_2O$) layer of P. serrulata var. spontanea leaves. By means of bioassay-directed fractionation, we isolated eleven known flavonoids (1-11) from the EtOAc soluble fraction of the MeOH extract of the Prunus serrulata var. spontanea leaves, exhibiting strong antioxidant activity and characterized as prunetin (1), genistein (2), quercetin (3), prunetin $4'-O-{\beta}-glucopyranoside$ (4), kaempferol $3-O-{\alpha}-arabinofuranoside$ (5), prunetin $5-O-{\beta}-glucopyranoside$ (6), kaempferol $3-O-{\beta}-xylopyranoside$ (7), genistin (8), kaempferol $3-O-{\beta}-glucopyranoside$ (9), quercetin $3-O-{\beta}-glucopyranoside$ (10) and kaempferol $3-O-{\beta}-xylopyranosyl-(1{\rightarrow}2)-{\beta}-glucopyranoside$ (11). Compounds 3 and 10 showed good activities in all tested model systems. Compounds 2 and 8 showed scavenging activities in the DPPH and $ONOO^-$ tests, while compounds 5, 7, 9 and 11 were active in the $ONOO^-$ and ROS tests. On the other hand, compounds 1, 4 and 6 did not show any activities in the tested model systems.

• Preventive Nutrition and Food Science
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• 제2권4호
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• pp.285-290
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• 1997

Previously, a 50% aqueous methanol extract of Galla rhois was shown to be the most potent tyrosinase inhibition activity with an {TEX}$IC_{50}${/TEX}(the concentration causing 50% inhibition of tyrosinase activity) of 0.2mg/ml of 205 crude drug extracts. To isolate tyrosinase inhibitors, the methanol extract was evaporated to a small volume in vacuo, and then partitioned stepwise with benzene and ethyl acetate(EtOAc). the EtOAc fraction was solubilized in 10% MeOH solution, and then fractionated successively by Diaion HP-20 and Sephadex LH-20 column chromatography, and preparative HPLC. Three phenolic compounds were isolated, and characterized as gallic acid(GA), methyl gallate(MG) and 1,2,3,4,6-penta-O-galloyl-$\beta$-D-glucose(PGG) by UV, IR, {TEX}${1}^H${/TEX}-&{TEX}${13}^C${/TEX}-NMR, and FAB-MS spectroscopy, PGG({TEX}$IC_{50}${/TEX}=50$\mu\textrm{g}$/ml) showed a considerable inhibitory effect against mushroom tyrosinase, while GA({TEX}$IC_{50}${/TEX}=1.6mg/ml) and MG({TEX}$IC_{50}${/TEX}=234$\mu\textrm{g}$/ml) did not show an appreciable effect. Meanwhile, MG inhibited greatly melanogenesis in a murine melanocyte cell line, Mel-Ab. MG and PGG showed typical noncompetitive inhibition patterns against mushroom tyrosinase. These results suggest that PGG and MG may be potentially useful as either anti-browning or anti-melanogenic agents in foods and cosmetics.

• Journal of Applied Biological Chemistry
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• 제58권2호
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• pp.109-112
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• 2015

Fruits of Rhus parviflora were extracted with 80% aqueous methanol (MeOH), and the concentrated extract was partitioned using ethyl acetate (EtOAc), n-butanol (n-BuOH), and $H_2O$, successively. Purification of EtOAc fraction led to isolation of fifteen polyphenols of which structures were identified by spectroscopic methods including 2D-NMR. Most compounds apart from compound 10 inhibited low density lipoproteinoxidation within $IC_{50}$ value of $10{\mu}M$. Among compounds, taxifolin (2), quercetin 3-O-${\alpha}$-L-rhamnopyranoside (13), agathisflavone (5) sulfuretin (4), and aureusidin (3) showed $IC_{50}$ values 0.9, 0.8, 5.8, 2.9, and $2.4{\mu}M$ which were of highly significant in comparison positive control butylated hydroxytoluene with $IC_{50}$ value of $2.1{\mu}M$. The results indicate fruits of R. parviflora as a source of antihypercholesterolemic compounds.

• Applied Biological Chemistry
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• 제51권4호
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• pp.316-320
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• 2008

과육을 제거한 산수유 씨의 MeOH 추출물의 EtOAc 분획물로부터 이용하여 10종의 화합물을 분리하였다. 이들 중에서 (+)- pinoresinol, (-)-balanophonin, vanillin, 4-hydroxybenzaldehyde, coniferaldehyde, betulinic acid는 이 식물에서 처음으로 분리된 물질들이다. 본 연구에서 분리된 10종의 화합물에 대한 in vitro 최종당화산물(AGEs) 생성억제효능 실험결과, (-)-balanophonin (화합물 2)과 gallicin(화합물 3)이 각각 $IC_{50}$ 값 27.81과 18.04${\mu}M$로 비교양성대조군인 aminoguanidine(974.59 ${\mu}M$)보다 훨씬 우수한 효능을 나타내었다.

• Archives of Pharmacal Research
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• 제26권4호
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• pp.279-285
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• 2003

In our ongoing study to identity antioxidants from natural sources, the antioxidant activity of Nelumbo nucifera stamens was evaluated for their potential to scavenge stable 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radicals, inhibit total reactive oxygen species (ROS) generation, in kidney homogenates using 2 ,7 -dichlorodihydrofluorescein diacetate (DCHF-DA), and scavenge authentic peroxynitrites ($ONOO^-$). A methanol (MeOH) extract of the stamens of N. nucifera showed strong antioxidant activity in the $ONOO^-$system, and marginal activity in the DPPH and total ROS systems, so were therefore fractionated with several organic solvents, such as dichloromethane ($CH_2 Cl_2$), ethyl acetate (EtOAc) and n-butanol (n-BuOH). The EtOAc soluble fraction, which exhibited strong antioxidant activity in all the model systems tested, was further purified by repeated silica gel and Sephadex LH-20 column chromatographies. Seven known flavonoids [kaempferol (1), kaempferol 3-Ο-$\beta$-D-glucuronopyranosyl methylester (2), kaempferol 3-Ο-$\beta$-D-glucopyranoside (3), kaempferol 3-Ο-$\beta$-D-galactopyranoside (4), myricetin 3 ,5 -dimethylether 3-Ο-$\beta$-D-glucopyranoside (5), kaempferol 3-Ο-$\alpha$-L-rhamnopyranosyl-(1$\rightarrow$6)-$\beta$-D-glucopyranoside (6) and kaempferol 3-Ο-$\beta$-D-glucuronopyranoside (7)], along with $\beta$-sitosterol glucopyranoside (8), were isolated. Compound 1 possessed good activities in all the model systems tested. Compounds 2 and 7 showed scavenging activities in the DPPH and $ONOO^-$ tests, while compounds 3 and 4 were only active in the $ONOO^-$ test. Conversely, compound 8 showed no activities in any of the model systems tested.

• Journal of Forest and Environmental Science
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• 제18권1호
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• pp.53-60
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• 2001

편백나무 잎을 채취하여 건조시킨 후 분말로 제조하여 아세톤-물(7:3, v/v)로 추출하고 헥산, 메틸렌클로라이드, 에틸아세테이트, 그리고 수용성으로 분획하여 동결건조 시켰다. 그 중에서 에틸아세테이용성 분획을 Sephadex LH-20으로 충진한 칼럼에서 메탄올과 에탄올-헥산 혼합액을 용매로 사용하여 칼럼크로마토그래피를 실시하였다. 단리된 화합물들은 박층크로마토그래피(TLC)로 확인한 후 NMR스펙트럼을 사용하여 정확한 구조규명을 하였고 FAB-MS스펙트럼으로 분자량을 측정하였다. 주로 많은 양의 taxifolin-3-O-${\beta}$-D-xylopyranoside와 (+)-catechin이 포함되어 있었으며 소량의 quercetin-3-O-${\alpha}$-L-rhamnopyranoside도 함께 단리 되었다.

• Journal of Applied Biological Chemistry
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• 제50권4호
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• pp.275-280
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• 2007

Dried unripe fruits of Rubus coreanus Miq. were extracted with 80% aqueous MeOH and the concentrated extract was partitioned with EtOAc and $H_2O$. From the EtOAc fraction, four triterpenoids were isolated through repeated silica gel, ODS and Sephadex LH-20 column chromatographies. From the result of physico-chemical data including NMR, MS aud IR, the chemical structures of the compounds were determined as tormentic acid (1), myrianthic acid (2), hovenic acid (3) and 2${\alpha}$,3${\beta}$,19${\beta}$,23-tetrahydroxylolean-12-en-28-oic acid (4). Compounds 3 and 4 were isolated for the first time from this plant. All isolated compounds were evaluated for cytotoxic activity against human colon carcinoma cells using in vitro three-[4,5-dimethylthiazole-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay, compound 3 showed a higher cytotoxicity ($IC_{50}$ = 7.8 ${\mu}M$) than doxorubicin ($IC_{50}$= 50 ${\mu}M$).