• Korean Journal of Acupuncture
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• v.25 no.4
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• pp.89-104
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• 2008

Objectives : This study was undertaken to determine the in vitro antioxidant activity of the extract of Chilgitang herb-acupuncture solution (CHAS). Methods : The radical scavenging capacity was tested by 2,2-diphenyl-1-picryl-hydrazyl (DPPH), hypoxanthine-xanthine oxidase system, DCFH-DA assay, nitric oxide and peroxynitrite generating system. In addition, antioxidant activity on copper and AAPH mediated human low-density lipoprotein (LDL) oxidation was measured by using TBARS assay and relative electrophoretic mobility assay. The amount of total phenolic compounds was assayed by the Folin-Ciocalteu method. Results : CHAS revealed a potent scavenging activity on DPPH radical(82%), superoxide anions(73%), hydroxyl radical(63%), nitric oxide (99%) and peroxynitrite (99%). Moreover, CHAS showed a strong inhibitory effect (59%) on $FeCl_2$-ascorbic acid induced lipid peroxidation of rat liver homogenate. CHAS also markedly inhibited copper(81%) and AAPH(56%)-mediated LDL oxidation, and effectively suppressed the electrophoretic mobility during exposure of human LDL to copper ions. CHAS (82 mg/g) contained higher concentration of total phenolic compounds than that of water extract (45 mg/g) obtained from Chilgitang. Conclusions : These results indicate that CHAS may protect against ROS- or RNS involved diseases, including cardiovascular diseases.

• Journal of Korean Medicine Rehabilitation
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• v.26 no.3
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• pp.51-58
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• 2016

Objectives This study was designed to investigate whether the Cheongajihwang-Tang (CT) has an inhibitory effect association with oxidation or inflammation in RAW264.7 cells. Methods Cytotoxic activity of CT extract on RAW264.7 cells was evaluated by using 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide (MTT) solution. Nitric oxide production was measured using Griess reagent system. The total phenolic contents and 1,1-Diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity was measured to evaluate the anti-oxidative effects of CT. Dichlorofluorescin diacetate (DCFH-DA) has been used as a substrate for measuring intracellular oxidant production. Results Cheongajihwang-Tang does not impair the cell viability in tested concentration. CT showed anti-oxidative effects in vitro by decreasing electron donating ability, and also showed anti-inflammatory effects suppressing NO and ROS expression in LPS induced RAW264.7 activation. CT inhibited the generation of intracellular ROS production as dose dependant manner. Conclusions CT has anti-oxidative effects and anti-inflammatory activities. These results indicate that CT extract has an anti-inflammatory activities via anti-oxidative effects.

• Journal of Life Science
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• v.20 no.9
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• pp.1365-1372
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• 2010

We investigated the inhibitory effects of solvent extracts from dried yam on $H_2O_2$-induced oxidative stress and growth of cancer cell lines (HT1080 human fibrosarcoma and HT-29 human colon cancer cells). Yam (Dioscoreacea) has been recognized as a healthy food due to its various biological activities, such as anti-obesity, anti-constipation, anti-proliferation, and anti-mutagenic activities, as well as its ability to decrease blood glucose and cholesterol levels. In order to determine the protective effect on $H_2O_2$-induced oxidative stress, DCFH-DA (dichlorodihydrofluorescin diacetate) assay was conducted. Acetone with methylene chloride (A+M) extract of dried yam appeared to reduce the levels of intracellular reactive oxygen species (ROS) with dose responses. Among the fractions, 85% aq. methanol fraction showed the highest protective effect on production of lipid peroxides. Inhibitory effects of A+M and methanol (MeOH) extracts on the growth of HT1080 and HT-29 cancer cells increased in a dose dependent manner. The treatments of n-hexane, 85% aq. methanol and n-butanol fractions (${\geqq}0.5$ mg/ml concentrations) significantly inhibited the growth of both cancer cells (p<0.05). From these results, 85% aq. methanol fraction showed inhibitory effects on cellular oxidation and growth of human cancer cells, suggesting that this fraction may contain active compounds of dried yam.

• Journal of Life Science
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• v.17 no.11
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• pp.1533-1538
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• 2007

This study was carried out to determine the inhibitory effects of solvent extracts from Sargassum hemiphyllum on growth of cancer cell lines (AGS human gastric adenocarcinoma and HT-29 human colon cancer cells) and production of lipid peroxides. Inhibitory effects of acetone with methylene chloride extract from S. hemiphyllum on the growth of AGS and HT-29 cancer cells were increased as dose dependent patterns (p<0.05). The methanol extract was more effective on inhibition of growth of AGS. The treatments of hexane, 85% aq. methanol, butanol and water fractions significantly inhibited the growth of cancer cells (p<0.05) and the inhibitory effect was stronger in HT-29. In DCFH-DA (dichlorodihydrofluorescin diacetate) assay, acetone with methylene chloride and methanol extracts showed a stronger inhibitory effect on the production of cellular lipid peroxides (p<0.05) compared with the butanol and hexane fractions. These results indicate that the consumption of S. hemiphyllum may be recommended as a potent functional food for preventing cellular oxidation and cancer.

• Herbal Formula Science
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• v.20 no.2
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• pp.65-82
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• 2012

Objectives : The aim of this study was verification of the anti-oxidative and anti-inflammatory effects of Odukhwan(ODH) and Sasinhwan(SSH) in mouse macrophage, RAW264.7 cells. Methods : To investigate the anti-oxidative effect and scavenging activities of DPPH radical, superoxide anions, nitric oxide and peroxynitrite were measured. Cytotoxic activity of extract of ODH and SSH on RAW264.7 cells was measured using MTS assay. To proof the reductive activity of intracellular oxidation, DCFH-DA assay was performed. The nitric oxide(NO) production was measured and pro-inflammatory cytokines and $PGE_2$ were measured by ELISA kit. The levels of inducible nitric oxide synthase(iNOS), cyclooxygenase-2(COX-2) and nuclear NF-${\kappa}B$ p65 expression were detected by western blot. Results : After those analyses, we bring to a conclusion as follows. Both herbal formulations scavenged DPPH radical and nitric oxide. But ODH had no scavenging activity of superoxide anions and SSH had low scavenging activity in peroxynitrite. And the results indicated that ODH and SSH inhibited the LPS-induced NO, $PGE_2$ production and iNOS, COX-2 expression accompanied by an attenuation of IL-$1{\beta}$ and IL-6 production in RAW264.7 cells. They also have suppression effects of LPS-induced NF-${\kappa}B$ activation. Conclusions : ODH and SSH have anti-oxidative and anti-inflammatory effects and they may be a part of database for development of new anti-oxidative and anti-inflammatory drugs.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.56 no.4
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• pp.349-360
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• 2011

The objectives of present study were to characterize the peptides which were isolated from Korean fermented soybean paste, chungkukjang, and to determine their antioxidant activities. Four fractions were collected from the methanol extract of chungkukjang by using a recycling preparative HPLC. Among fractions, Fr-2 was identified to be highly potent free radical scavenging activity in the assay of 1,1-diphenyl-2-picryl-hydrazyl (DPPH) and nitroblue tetrazolium(NBT)-reduction inhibition. Base on antioxidant effects, fraction Fr-2 was employed for the refraction with a prep-column and separated into five fractions of which two fractions were identified to have higher antioxidant activity. To confirm the amino acid constituents of antioxidant fractions Fr-2-2 and Fr-2-3 were analyzed, and eight kinds of amino acids such as aspartic acid, threonine, serine, glutamic acid, glycine, lysine, histidine, and arginine were identified as the constituent amino acids. Antioxidant activities of the separated peptides were further assessed cell viability with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl terazolium bromide (MTT), and fluorescence-activated cell sorting (FACS) analysis of H4IIE cells treated with hydrogen peroxide (H2O2). Chungkukjang peptides have shown their ability to protect H4IIE rat hepatoma cells against H2O2- induced oxidative stress by concentration and time-dependent manner. Therefore, These results indicated that fermented soybean paste chungkukjang will be promoted the antioxidant and radical scavenging activities, and beneficial for health. The antioxidant peptide fractions Fr-2-2 and Fr-2-3 were denominated as P-NICS-1 and P-NICS-2, respectively. However, further studies were required to clarify their amino acid sequences and molecular properties, and physiological significances.

• Food Science and Preservation
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• v.18 no.1
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• pp.65-71
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• 2011

We explored the effect of extracts of dried Platycodon grandiflorum on production of reactive oxygen species (ROS), glutathione (GSH) and nitric oxide (NO). To determine antioxidant activity in the presence of $H_2O_2$-induced oxidative stress, DCFH-DA (dichlorodihydrofluorescin diacetate) assay was employed. Acetone/methylene chloride (A+M) and methanolic (MeOH) extracts of P. grandiflorum reduced intracellular ROS levels. Of the various tested fractions, n-BuOH fraction showed the highest protective effect in terms of lipid peroxide production. Total GSH levels were measured after treatment of HT1080 cells with the A+M and MeOH extracts, and other solvent fractions, at various concentration. The A+M extacts and 85% (v/v) aqueous MeOH fraction significantly increased GSH levels (p<0.05). When lipopolysaccharide (LPS)-induced NO production was evaluated, all tested crude extracts, and fractions thereof, significantly reduced NO production (p<0.05), and the n-BuOH and 85% (v/v) aqueous MeOH fractions (at 0.05 mg/mL) showed the strongest inhibitory effects. The results showed that the n-BuOH fraction inhibited both cellular oxidation and NO production, and this fraction may thus contain valuable active compounds.

• Journal of Life Science
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• v.26 no.8
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• pp.948-954
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• 2016

We investigated the flavonoid and phenol contents and antioxidant effect of wine by-product extract. Antioxidant effects were measured with 1,1-diphenyl-2-picryhydrazyl (DPPH) and 2.2'-azino-bis(3-ethylbenothiazoline-6-sulfonic acid) diammonium salt radical cation (ABTS+) assays. Cellular reactive oxygen species (ROS) were measured with the dichlorofluorescein-diacetate (DCFH-DA) assay. The flavonoid and phenol contents of the methanol (MeOH) extract were greater than those of the acetone+methylene chloride (A+M) extract. Among fractions, the 85% aqueous methanol (85% aq. MeOH) fraction contained the highest flavonoid contents, while the n-BuOH fraction had more phenol contents. In the DPPH and ABTS assays, the MeOH extract showed a scavenging effect greater than that of the A+M extract (p<0.05). The n-BuOH fraction (0.5 mg/ml concentrations) showed scavenging effects of 72% and 92%, respectively, in the DPPH and ABTS assays (p<0.05). However, the 85% aq. MeOH fraction showed a 90% scavenging effect in the DPPH assay only. In 120 min ROS production assay, all tested fractions dose-dependently decreased cellular ROS production induced by H₂O₂ in comparison with that produced by exposure to the extract-free control. The MeOH extract showed a higher sinhibitory effect on cellular ROS producing than that of the A+M extract at all concentrations tested. Treatment with the n-BuOH fraction (0.1 mg/ml concentrations) inhibited cellular ROS production by 60%. These results indicate that the n-BuOH fraction of wine by-product extract inhibited cellular oxidation and may contain valuable bioactive compounds such as flavonoids and phenols.

• Journal of Life Science
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• v.27 no.7
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• pp.796-804
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• 2017

This study was carried out to evaluate the antioxidant effect of methanolic extract of Houttuynia cordata (HCME) and to identify a compound having antioxidant effect. The ethyl acetate fraction of HCME showed the highest antioxidant effect in organic solvent fractions. The fraction was then separated into 12 fractions by open column chromatography. Among these fractions, the fraction 10 (Fr. 10) with the highest antioxidant activity was isolated, and its antioxidant effect was evaluated by DPPH radical scavenging activity, reducing power, TBARS, cell viability, DNA oxidation and DCF fluorescence. The Fr. 10 at a $64{\mu}g/ml$ showed 60% of inhibitory effect similar to that of vitamin C at $10{\mu}g/ml$, compared with blank group. The Fr. 10 at $64{\mu}g/ml$ showed 264% of reducing power, compared with blank group. TBARS assay showed that the Fr. 10 at $64{\mu}g/ml$ had 35.5% of inhibitory effect similar to that of vitamin E at $1,000{\mu}g/ml$, compared with blank group. The Fr. 10 above $32{\mu}g/ml$ displayed cytotoxicity. However, it was observed that the Fr. 10, above $1{\mu}g/ml$ reduced DNA damage. DCF fluorescence assay showed that the Fr. 10 inhibited oxidative stress by $H_2O_2$ in a dose dependent manner. The compound of Fr. 10 was identified to be rutin whose molecular weight is 610 by the IR and LC-MS analyses. Therefore, these results suggest that the rutin of Fr. 10 could use as a natural antioxidant for development of cosmetics and functional foods.