• 한국수산과학회지
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• 제53권1호
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• pp.123-131
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• 2020

In this study, we investigated the protective effects of a Neutrase enzymatic hydrolysate derived from Korea pen shell Atrina pectinata (APN) against hydrogen peroxide (H₂O₂)-induced oxidative damage in hepatocytes. First, we confirmed that APN has antioxidant activities by scavenging 2,2-azino-bis (3-ethylbenzthiazoline)-6-sulfonic acid radical (ABTS⁺) and H₂O₂ and increasing oxygen radical absorbance capacity (ORAC) value. Also, the treatment of APN increased the cell viability by reducing the intracellular reactive oxygen species (ROS) production in H₂O₂-stimulated hepatocytes. In addition, APN decreased the sub-G₁ DNA contents and the apoptotic body formation increased by H₂O₂ stimulation. Moreover, APN modulated the protein expression of apoptosis related molecules (Bcl-2, Bax and p53) by suppressing the activation of nuclear factor NFkB and ERK/p38 signaling in H₂O₂-stimulated hepatocytes. Furthermore, APN led to the activation of Nrf2/HO-1signaling known as antioxidant systems. These results suggest APN protects hepatocytes against oxidative damages caused by H₂O₂ stimulation.

• 대한한의학회지
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• 제30권5호
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• pp.146-156
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• 2009

Objectives: The objective of this study was to investigate the antioxidant effects of manufactured Wen-pi-tang-Hab-Wu-ling-san (WHW$^{(R)}$) in vitro. Methods: WHW$^{(R)}$ was prepared by the pilot manufacture of WHW water extract from a GMP system appointed company. Antioxidative activities were determined by in vitro tests as follows: the scavenging activities of oxygen free radicals including 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical, superoxide anion, hydrogen peroxide and nitric oxide radicals, as well as ferrous ion chelating capacity and Trolox equivalent antioxidant capacity (TEAC). Results: WHW$^{(R)}$ significantly scavenged oxygen free radicals such as DPPH (IC$_{50}$=115.28 $\pm$ 0.25 $\mu$g/$m\ell$), superoxide anion (IC$_{50}$=8.56 $\pm$ 0.08 $\mu$g/$m\ell$), hydrogen peroxide (IC$_{50}$=240.36 $\pm$ 3.41 $\mu$g/$m\ell$) and nitric oxide (IC$_{50}$=162.28 $\pm$ 0.21 $\mu$g/$m\ell$) radicals. WHW$^{(R)}$ also showed ferrous ion chelating activity (IC$_{50}$=543.19 $\pm$ 4.85 $\mu$g/$m\ell$) and Trolox equivalent effects (IC$_{50}$=45.311 $\mu$g/$m\ell$) in TEAC and ORAC assay, respectively. Conclusion: This study demonstrates that WHW$^{(R)}$ has strong antioxidative properties through free radical scavenging activity. These data suggest that WHW$^{(R)}$ be used as an antioxidant agent.

• Reproductive and Developmental Biology
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• 제35권4호
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• pp.485-489
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• 2011

The present study was conducted to examine the reactive oxygen species (ROS) generation levels and subsequent DNA damage in the bovine cultured somatic cells. Bovine ear skin cells were classified by serum starvation, confluence and cycling cells. Cells were stained in 10 ${\mu}M$ dichlorohydrofluorescein diacetate ($H_2DCFDA$) or 10 ${\mu}M$ hydroxyphenyl fluorescein (HPF) dye to measure the $H_2O_2$ or $^{\cdot}OH$ radical levels. The samples were examined with a fluorescent microscope, and fluorescence intensity was analyzed in each cell. $H_2O_2$ and $^{\cdot}OH$ radical levels of cultured somatic cells were high in confluence group ($7.1{\pm}0.7$ and $8.4{\pm}0.4$ pixels/cell, respectively) and significantly low in serum starvation group ($4.9{\pm}0.4$ and $7.0{\pm}0.4$ pixels/cell, respectively, p<0.05). Comet tail lengths of serum starvation ($148.3{\pm}5.7$ ${\mu}M$) and confluence ($151.1{\pm}5.0$ ${\mu}M$) groups were found to be significantly (p<0.05) increased in comparison to that of cycling group ($137.1{\pm}7.5$ ${\mu}M$). These results suggest that the culture type of donor cells can affect the ROS generation, which leads the DNA fragmentation of the cells.

• 대한수의학회지
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• 제37권3호
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• pp.619-627
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• 1997

It is known that 2-methyl-1,4-naphtoquinone(menadione, MD) induces hepatotoxicities both in vivo and in vitro. These toxic effects are believed to result from oxidative damages to hepatocytes by "active oxygen" species via one-electron reduction of the naphtoquinone. The ginsenoside(GS) is a complex mixture of individual ginsenosides which is known to produce a range of effects on the cardiovascular and central nervous systems. In particular, GS has an antioxidant effect. In this experiment we studied the effect of GS from red panax ginseng(red ginseng total saponin, RGTS) on free radical-induced liver injuries by MD. Administration of MD($150{\mu}M$) caused an increase in aspartate aminotransferase(AST) activities and lipid peroxidation, decrease in alkaline phosphatase(ALP) activities and total bilirubin levels in blood, caused depletion of GSH and changes of antioxidant enzyme(superoxide dismutase, catalase) activities are shown in liver tissue. Administration of RGTS restored the AST levels that increased by MD, but catalase showed no significant changes. RGTS also had an effect of restoring the GSH level and had some synergistic effects with SOD. These data suggest that RGTS may have some protective effects on liver injury which is related with the oxygen free radical.

• 대한화장품학회지
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• 제32권4호
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• pp.263-267
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• 2006

태양광선과 산소는 피부세포에 자유라디칼(free radical)과 활성산소종(reactive oxygen species, ROS)을 생성시켜 세포막 지질의 과산화와 염증을 유발한다. 본 연구는 자외선에 의한 피부손상에 대한 머위추물물의 피부 보호효과에 대한 것으로 UVB가 조사된 각질형성세포에서이 항산화 및 염증관련 사이토카인의 억제효과를 측정하였다. 라디칼에 의한 지질과산화에 대한 억제효과는 $100{\mu}g/mL$에서 70.1%로 우수하게 나타났으며, 세포내에서 ROS에 의해 형광을 띠는 물질로 전환되는 $CM-H_2DCFDA$를 이용하여 ROS의 양을 측정한 결과 자외선에 의해 증가된 세포내 ROS의 양이 머위추출물을 처리함으로써 $500{\mu}g/mL$ 농도에서 45% 이상의 우수한 소거효과를 나타냈다. 또한 각질형성세포에서 UVB에 의해 생합성이 증가되는 $IL-1{\alpha}$ 및 $PGE_2$의 생합성 억제효과는 $100{\mu}g/mL$에서 각각 25.7%, 59.3%를 저해하는 것으로 우수한 효과를 나타내었다. 따라서 머위추출물은 각질형성세포에서 자외선 조사에 의한 피부손상에 대한 보호효과를 가진 화장품에서 우수한 소재로 적용될 수 있다.

• Restorative Dentistry and Endodontics
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• 제26권5호
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• pp.399-408
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• 2001

Tooth bleaching has been prevailing recently for its ability to recover the color and shape of natural teeth without reduction of tooth material. However, it has been reported that bleaching procedure adversely affects the adhesive bond strength of composite resin to tooth. At the same time the bond strength was reported to be regained by application of some chemical agents. The purpose of this in vitro study was to investigate the effect of the removal of residual peroxide on the composite- enamel adhesion and also evaluated fracture mode between resin and enamel after bleaching. Sixty extracted human anterior and premolars teeth were divided into 5 groups and bleached by combined technique using of office bleaching with 35 % hydrogen peroxide and matrix bleaching with 10% carbamide peroxide for 4 weeks. After bleaching, the labial surfaces of each tooth were treated with catalase, 70% ethyl alcohol, distilled water and filled with composite resin. Shear bond strength was tested and the fractured surfaces were also examined with SEM. Analysis revealed significantly higher bond strength values. (p＜0.05) for catalase-treated specimens, but water-treated specimens showed reduction of bond strength, alcohol- treated specimens had medium value between the two groups(p＜0.05). The fracture mode was shown that the catalase group and the alcohol group had cohesive failure but the water sprayed group had adhesive failure. It was concluded that the peroxide residues in tooth after bleaching seems to be removed by gradual diffusion and the free radical oxygen from peroxide prevents polymerization by combining catalyst in the resin monomer. Therefore it may be possible to eliminate the adverse effect on the adhesion of composite resin to enamel after bleaching by using water displacement solution or dentin bonding agent including it for effective removal of residual peroxide.

• 한국식품조리과학회지
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• 제33권1호
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• pp.113-120
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• 2017

Purpose: Lipid autoxidation of a soybean oil-in-water emulsion with high oil content was studied under after basil extract and/or iron addition. Methods: The emulsion consisted of tocopherol-stripped soybean oil (40 g), citrate buffer (60 g, pH 4.0), and/or $FeSO_4$ (0.5 mg) with 75% ethanol extract (200 mg/kg) of basil (Ocimum basilicum). Lipid oxidation was evaluated using headspace oxygen content, hydroperoxide contents, and p-anisidne values of the emulsion. Polyphenol compound retention in the emulsion during oxidation was determined spectrophotometrically. Results: Addition of basil extract significantly (p<0.05) decreased reduced hydroperoxide contents of the emulsion, and iron significantly (p<0.05) increased anisidine values and decreased oxygen contents. Co-addition of basil extract and iron showed significantly (p<0.05) lower reduced hydroperoxide contents in the emulsion than compared to those of the emulsion with added iron and the control emulsion without basil extract nor or iron. During the emulsion oxidation, polyphenol compounds in the emulsion with added basil extract were degraded, but more slowlywhich was slowed degraded in the presence of iron. Conclusion: The iIron increased the lipid oxidation through hydroperoxide decomposition, and basil extract showed antioxidant activity through radical-scavenging and iron-chelation. Polyphenol degradation was decelerated by iron addition, which suggested suggests iron chelation may be more preferred topreferentially activated over radical scavenging in the antioxidant action by of basil extract in the oil-in-water emulsion with high oil content.

• Environmental Analysis Health and Toxicology
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• 제13권3_4호
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• pp.105-115
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• 1998

Reactive oxygen species (ROS) are highly reactive molecules due to their unpaired electron. They have been suspected as one of the major tissue damage inducers in biological metabolic systems. Antioxidant enzymes, such as catalase and superoxide dismutase, could not repair all the oxidative damages resulting from those excessive toxic ROS. It is, therefore, urgent to develop effective antioxidants to relieve from the oxidatire damages. In this study antioxidative effects were investigated by using two flavonoids such as quercetin and naringenin and a flavonoid-rich extract, Ginkgo biloba extract in combination with paraquat that is known as a strong generator of oxygen radicals. The results are summeringed as follows: 1. To assess radical scavenging ability reduction concentrations (IC$_{50}$) of 1,1-diphenyl-2-picrylhydrazine (DPPH) within 15 minutes were measured. The values of the IC$_{50}$ of quercetin and Ginkgo biloba extract were 15.4 $\mu$M and 13.2$\mu$g/ml, respectively. Their radical removing activities showed concentration-dependent manners. 2. In the hydrogen peroxide assay by using PMS-NADH system, quercetin, naringenin and Ginkgo biloba extract led to removing hydrogen peroxide in concentrationdependent manner whose removing abilities at 100$\mu$M or 100 $\mu$g/ml were 75.6, 25.8 and 26.0%, respectively. 3. In the hydrogen peroxide-induced rat blood hemolysis assay all three compounds led to similar effects whose hemolysis inhibition ratios at 100$\mu$M or 100$\mu$g/ml were 68.0, 5.14 and 55.8%, respectively. 4. In the xanthinee oxidase assay by measuring degree of NADH oxidation in the presence of hypoxanthine and xanthinee oxidase, both quercetin and Ginkgo biloba extract showed excellent activities showing 42.8 and 24.2% inhibiting xanthine oxidase activity at 100$\mu$M or 100$\mu$g/ml concentrations, respectively.

• Archives of Pharmacal Research
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• 제26권10호
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• pp.809-815
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• 2003

Since reactive oxygen species (ROS) and hydroxyl radicals ($^-OH$) play an important role in the pathogenesis of many human degenerative diseases, much attention has focused on the development of safe and effective antioxidants. Preliminary experiments have revealed that the methanol (MeOH) extract of the stem of Prunus davidiana exerts inhibitory/scavenging activities on 1, 1-diphenyl-2-picrylhydrazyl (DPPH) radicals, total ROS and peroxynitrites ($ONOO^-$). In the present study, the antioxidant activities of this MeOH extract and the organic solvent-soluble fractions, dichloromethane (CH$_2$Cl$_2$), ethyl acetate (EtOAc), and n-butanol (n-BuOH), and the water layer of P. davidiana stem were evaluated for the potential to inhibit $^-OH$ and total ROS generation in kidney homogenates using 2',7'-dichlorodihydrofluorescein diacetate (DCHF-DA), and for the potential to scavenge authentic $ONOO^-$. We also evaluated the inhibitory activity of seven flavonoids isolated from P. davidiana stem, kaempferol, kaempferol 7-Ο-$\beta$-D-glucoside, (+)-catechin, dihydrokaempferol, hesperetin 5-Ο-$\beta$-D-glucoside, naringenin and its 7-Ο-$\beta$-D-glucoside, on the total ROS, $^-OH$ and $ONOO^-$ systems. For the further elucidation of the structure-inhibitory activity relationship of flavonoids on total ROS and 'OH generation, we measured the antioxidant activity of sixteen flavonoids available, including three active flavonoids isolated from P. davidiana, on the total ROS and 'OH systems. We found that the inhibitory activity on total ROS generation increases in strength with more numerous hydroxyl groups on their structures. Also, the presence of an ortho-hydroxyl group, whether on the Aring or S-ring, and a 3-hydroxyl group on the C-ring increased the inhibitory activity on both total ROS and $^-OH$ generation.

• Preventive Nutrition and Food Science
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• 제10권4호
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• pp.316-325
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• 2005

The antioxidant potential of a $75\%$ methanolic extract of highbush blueberry (Vaccinium corymbosum L.) and its different fractions was investigated using different reactive oxygen species (ROS), nitric oxide (NO.), metal chelating and lipid peroxidation assays. Methylene chloride and $75\%$ methanol fractions showed equally high activities $(IC_{50} 0.010 mg/mL)$ for hydroxyl radical (HO) scavenging. Higher hydrogen peroxide $(H_2O_2)$ scavenging values were reported for the ethyl acetate and methylene chloride fractions and their $IC_{50}$ values were 0.20 and 0.15 mg/mL, respectively. Nitric oxide (NO.) and 1,1-diphenyl-2-picrylhydrazyl radical (DPPH) scavenging activities were higher in ethyl acetate and methylene chloride fractions. Chloroform and water fractions showed higher activities in superoxide $(O_2.)$ scavenging. All fractions showed strong metal chelating capacities compared with the commercial antioxidants tested. The $0.1\%$ ethyl acetate fraction showed notable capacity to suppress lipid peroxidation in both fish oil and linoleic acid. Phenolic content was measured in all the fractions and methanolic extract. Among the fractions, ethyl acetate fraction showed the highest phenolic content.