• Journal of Life Science
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• v.32 no.4
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• pp.310-317
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• 2022

Recently, interest in the harmful factors of particulate matter (PM), a major component of air pollution, has been increasing. In particular, PM_2.5 with a diameter of less than 2.5 ㎛ is well known to induce oxidative stress accompanied by autophagy in human lung epithelial cells. However, studies on whether PM_2.5 increases autophagy under oxidative stress and whether this process is reactive oxygen species (ROS)-dependent are insufficient. Therefore, in this study, we investigated whether PM_2.5 promotes autophagy through the generation of ROS in human alveolar epithelial A594 cells. According to our results, cells co-treated with PM_2.5 and hydrogen peroxide (H₂O₂) showed a lower cell viability than cells treated with each alone, which was associated with increased total and mitochondrial ROS production. The co-treatment of PM_2.5 and H₂O₂ also increased autophagy induction, which was confirmed through Cyto-ID staining, and the expression of autophagy biomarker proteins increased. However, when ROS generation was artificially blocked by N-acetyl-L-cysteine pretreatment, the reduction in cell viability and induction of autophagy by PM_2.5 and H₂O₂ co-treatment were markedly attenuated. Therefore, the present results suggest that PM_2.5-induced ROS generation may play a critical role in autophagy induction in A549 cells.

• Archives of Pharmacal Research
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• v.25 no.6
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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.

• Journal of Pharmacopuncture
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• v.13 no.3
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• pp.5-13
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• 2010

Reactive Oxygen Species(ROS) are continuously produced at a high rate as a by-product of aerobic metabolism. Since tissue damage by free radical increases with age, the reactive oxygen species(ROS) such as hydrogen peroxide($H_2O_2$), nitric oxide(NO). Several lines of evidence provided that ROS appears to cause to develop aging-related various diseases such as cancer, arthritis, cardiovascular disease. Our reserch objective was to examine the in vitro biological activity of Scolopendrid Pharmacopuncture, including the total poly-phenol content, DPPH radical scavenging, ABTS radical scavenging, Superoxide dismutase(SOD)-like activity, Nitrite scavenging ability. The total poly-phenol contents of Scolopendrid Pharmacopuncture was 35.859mg/L. Elctron donation ability on DPPH was 36.82%. The 2,2'-azinobis-3-ehtlbezothiazoline-6-sulfonic acid radical decolorization (ABTS) was 84.7%. The superoxide dismutase (SOD)-like activities of Scolopendrid Pharmacopuncture was 44.33%. The nitrite scavenging effects were pH dependent, and were highest at pH 1.5(45.2%) and lowest at pH 6.0(11.3%). We conclude that Scolopendrid Pharmacopuncture may be useful as potential sources of antioxidant.

• Archives of Pharmacal Research
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• v.26 no.4
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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.

• The Korea Journal of Herbology
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• v.23 no.3
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• pp.19-25
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• 2008

Objectives : This study was performed to evaluate the neuroprotective effect of water extracts from Thujae Semen(TSW) in PC12 cells. Methods : We performed 2,2-diphenyl-2-picrylhydrazyl(DPPH) radical scavenging assay, 2,2-azinobis- (3-ethyl-benzothiazoline-6-sulfonic acid(ABTS) cation scavenging assay, and determination of total polyphenolic content to examine the antioxidant effects of TSW. We also carried out 3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay(MTT), reactve oxygen species(ROS) assay, and nitric oxide(NO) assay to examine neuroprotective effects against 6-hydroxydopamine(6-OHDA) in PC12 cells. Results : TSW showed $IC_{50}$ values of 404.3 and 219.9 ${\mu}g/mL$ in DPPH and in ABTS assays, respectively. TSW showed 9.74 ${\mu}g/mL$ of total polyphenol contents. TSW incresed cell viability in a dose dependent manner and it showed protective effect against 6-OHDA neurotoxicity at the concentration of 25-200 ${\mu}g/mL$. Moreover, it recovered 6-OHDA induced cell death at the same concentrations. The extract showed a dose dependent reduction of ROS and NO generation by 6-OHDA. Conclusions : We concluded that TSW has neuroprotective effect against 6-OHDA-induced toxicity in PC12 cells through ROS and NO inhibition.

• The Korea Journal of Herbology
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• v.23 no.3
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• pp.53-60
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• 2008

Objectives : Oxidative stress has a critical role in neurodegenerative diseases. In this study, we investigated the antioxidant and neuroprotective effects of the ethanolic extract of Banryong-hwan (BRHE) in SH-SY5Y cells and primary rat mesencephalic dopaminergic neurons. Methods : To assess the antioxidant effects, we carried out 1,1-diphenyl-2-picrylhydrazyl(DPPH) free radical scavenging assay, 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid)(ABTS) radical cation decolorization assay, and determination of total polyphenolic content. We evaluated the effect of BRHE treatment on neuroprotection against 6-hydroxydopamine(6-OHDA) toxicity using thiazolyl blue tetrazolium bromide(MTT) assay, nitric oxide(NO) assay, reactive oxygen species(ROS) assay in SH-SY5Y cells and tyrosine hydroxylase(TH) immunocytochemistry in primary rat mesencephalic dopaminergic neurons. Results : BRHE showed IC50 values of 328.10 ${\mu}g/mL$ and 43.12 ${\mu}g/mL$ in DPPH assay and in ABTS assay, respectively. Total polyphenolic content was 180.76 ${\mu}g/mL$. In SH-SY5Y cells, BRHE significantly attenuated the toxicity induced by 6-OHDA at the concentrations of 25-100 ${\mu}g/mL$ pre- and post- treatment in MTT assay. While 6-OHDA increased the NO and ROS contents, BRHE decreased them in a dose dependent manner. Moreover, in primary dopaminergic neuron culture, BRHE significantly protect-ed the dopaminergic cell loss against 6-OHDA toxicity up to 136% at the concentration of 75 ${\mu}g/mL$. Conclusions : These results demonstrate that BRHE has neuroprotective effect against 6-OHDA induced neurotoxicity through decreasing NO and ROS generation.

• The Korean Journal of Food And Nutrition
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• v.34 no.5
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• pp.423-429
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• 2021

Barley's nutritional value as a health food is increasing due to its excellent nutritional functionality. In this study, the levels of β-glucan, total polyphenols, and total flavonoids were analyzed in the ethanol extracts of different barley cultivars (Hinchalssal, Heuksoojeongchal, Betaone, Ganghochung, and Saechalssal). Also, the free radical scavenging abilities of 2,2-diphenyl-1-picryl-hydrazil (DPPH) and 2,2'-azino-bis-3-ethylbenzo-thiaxoline-6-sulfonic acid (ABTS) were measured to determine their antioxidant activity. The results confirmed that Betaone extract contained highly activefunctional components and exhibitedantioxidant activity. Next, we evaluated the hepatoprotective and inhibitory effects of reactive oxygen species (ROS) generated by barley ethanol extracts after inducing oxidative stress with tert-butyl hydroperoxide (tBHP) in HepG2 cells. Hinchalssal and Saechalssal extracts showed the most significant cytoprotective effect and also reduced ROS production significantly. These results suggest that Hinchalssal, Saechalssal, and Betaone represent potential natural antioxidant and hepatoprotective agents.

• The Journal of Internal Korean Medicine
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• v.43 no.3
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• pp.344-362
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• 2022

Objective: In this study, the antioxidant activity of Ojayeonjong-hwan extracts was compared, and the following results were obtained. Methods: For hydrothermal and ethanol extracts, DPPH free radical and ABTS cationic radical erasing activity and reducing power using the FRAP method were compared, and the association between the antioxidant power of each extract and total phenol content was investigated. Significant results were obtained through in vitro apoptosis analysis through FFITC staining, mitochondrial membrane potential analysis, and ROS level measurement using C2C12 myoblastoma. Results: 1. In a comparison of DPPH free radical and ABTS cationic radical scavenging activity, water, and 70% ethanol extracts of Ojayeonjong-hwan (WEO and EEO) showed superior radical scavenging ability. 2. In the results of reducing power using the FRAP method, WEO and EEO showed antioxidant activity, which was shown to be dependent on the total phenol content contained in the extracts. 3. In comparison to the protective effect against H₂O₂-induced oxidative stress in C2C12 myoblasts, water extracts had no significant effect, but 70% ethanol extracts inhibited H₂O₂-mediated cytotoxicity in a concentration-dependent manner. 4. The cytotoxic protective effect of EEO against oxidative stress in C2C12 myoblasts was correlated with its inhibitory effects on H₂O₂-induced apoptosis and cell-cycle arrest. 5. In H₂O₂-treated C2C12 myoblasts, the apoptosis inhibitory effects of EEO were associated with the suppression of mitochondrial dysfunction and DNA damage. 6. The protective effects of EEO against H₂O₂-induced oxidative stress in C2C12 myoblasts were directly related to the inhibition of ROS generation. Conclusions: Ojayeonjong-hwan extracts all have protective potential against oxidative stress.

• Journal of the Korean Applied Science and Technology
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• v.34 no.3
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• pp.568-576
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• 2017

As interest in functionality and environmentally friendly cosmetics is growing in recent years, materials that use safe and effective plant extracts have been developed. Therefore, this study also attempted to check the possibility of the graviola extract, which is known to have various efficacy mainly as a health functional material as a functional cosmetic material. In order to find out the antioxidant activity of graviola, we measured total polyphenol, total flavonoid content and DPPH radical scavenging activity and measured the ROS activity inhibition effect and cytoprotective effect on oxidative stress by treating HDF with hydrogen peroxide cells at an appropriate concentration after checking cytotoxicity in HDF cells. Based on the results of this experiment, the graviola extract was found to contain as high as 26.6 mg(CA)/100g, 14.3 mg(Q)/100g of total polyphenol and flavonoid, which are the antioxidant indexes and to have the high radical scavenging activity. The cell survival rate of the HDF cells was measured, and as a result, no significant cytotoxicity was observed at all concentrations and the experiment was carried out at a concentration of $100{\mu}g/mL$ afterwards. Inhibition of ROS activity in HDF cells induced by hydrogen peroxide was measured and the concentration-dependent inhibition of ROS activity was found and the cell protection effect of graviola was measured after hydrogen peroxide was treated for 4, 24 and 48 hours. As a result, the cell protection effect as high as 89.92% was confirmed at a $25{\mu}g/mL$ concentration up to 24 hours. As these results show that the graviola extract has excellent antioxidant activity, almost no toxicity to HDF cells, an effective activity inhibitory effect on active oxygen generated by hydrogen peroxide and excellent cytoprotective effect, the possibility as various functional materials with antioxidant and cytoprotective effects was confirmed.

• Journal of the East Asian Society of Dietary Life
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• v.19 no.6
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• pp.987-995
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• 2009

The effects of Kwamaegi on serum lipid profiles and ROS(reactive oxygen spices) generating and scavenging enzyme activities were investigated in rats. The three experimental groups were divided as follows: normal control diet group (NC), 5% naturally prepared and freeze-dried Kwamaegi supplemented diet group (NPK) and 5% chitosan-ascorbate treated and artificially dried (CWDD: Chilly Wind & Dehumidification Drier) Kwamaegi supplemented diets group (CAK). There were no significant differences in weight gain, feed intake, feed efficiency ratio or organs weights per body weight including liver, kidney, heart and spleen among the group. In addition, there were no significant differences in serum triglyceride and total cholesterol contents. The HDL-cholesterol contents of the NC, CAK and NPK groups were 62.00, 36.48 and 78.44 mg/dL while LDL-cholesterol contents were 62.00, 36.48 and 78.44 mg/dL, respectively, which were significantly different. The atherogenic indeces in the experimental groups were 0.62, 1.20 and 0.13, respectively. There were no significant differences in total XOD (xanthine oxidoreductase) activities; however XOD type O activity was higher in the NPK group than un the NC group and in the CAK group XOD type O activity was 21~45% lower compared to NC and NPK groups. SOD (superoxide dismutase) activity was significantly higher in the CAK group than in the NC and NPK groups, while there were no significantly differences in GST (glutathione S-transferrase) activity among the groups. Furthermore, serum ALT activity was higher in the NPK group versus the NC and CAK groups. GSH (glutathione) content was higher and LPO (lipid peroxide) content lower in the CAK group compared to the NC and NPK groups. Forem the above results, we suggest that CA treated and artificially dried Kwamaegi is not only a hygienic product but also has lowering effects on LDL-cholesterol and the atherogenic index together with the lowering of ROS-generating and increasing of ROS-scavenging enzyme activities compared to other natural products.