• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.1
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• pp.13-18
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• 2006

The antioxidant activity of the extract of Erigeron annuus was assessed by means of two different in vitro tests: bleaching of the stable 1,1-diphenyl-2-picrylhydrazyl radical (DPPH test) and the scavenging of authentic peroxynitrite in company with peroxynitrite generation from 3-morpholinosydnonimine (SIN-1). In both tests, the 85% aq. MeOH and n-BuOH soluble fractions of the crude extract showed a significant scavenging effect on peroxynitrite and DPPH radical in comparison to L-ascorbic acid. And bioassay-guided fractionation of the n-BuOH soluble fraction led to the isolation of three compounds: Apigenin (1), quercetin-3-O-glucoside (2), and caffeic acid (3). The structures of the isolated compounds were elucidated on the basis of their spectroscopic data and their antioxidant activities were measured by determining their capacity to scavenge peroxynitrite and the DPPH radical.

• Biotechnology and Bioprocess Engineering:BBE
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• v.9 no.3
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• pp.212-216
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• 2004

Peroxynitrite formation in vivo is implicated in numerous human diseases and there is considerable interest in the use of antioxidants and natural products for their treatment. The three components (1-3) isolated from Sargassum thunbergii as well as the organic solvent-soluble fractions and the aqueous layer of S. thunbergii were evaluated for their potential to scavenge authentic ONOO$\^$-/ and ONOO$\^$-/ derived from 3-morpholinosydnonimine (SIN-1). The antioxidant activity of the individual fractions was in the order of 85% aqueous (aq.) MeOH＞n$\^$-/ BuOH＞n-hexane＞H$_2$O. The three known compounds, sargahydroquinoic acid (1), sargaquinoic acid (2) and sargachromenol (3) showed peroxynitrite-scavenging activities comparable to those of L-ascorbic acid and penicillamine. These results showed a possible antioxidant activity in major constituents of S. thunbergii.

• Natural Product Sciences
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• v.5 no.2
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• pp.80-84
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• 1999

Peroxynitrite is a cytotoxic intermediate produced by the reaction between the superoxide anion radical and nitric oxide. Flavonoids (afzelin, quercitrin and quercetin 3-O-sambubioside), and chlorogenic acid and its methyl ester obtained from leaves of loquat (Eriobotrya japonica) have recently been shown to scavenge 1,1-diphenyl-2-picrylhydrazyl radical and to inhibit lipid peroxidation in mouse liver homogenate. The aim of this study is to investigate the inhibitory effects of the above components on peroxynitrite produced stimulated by 3-morpholinosydnonimine (SIN-1) to produce superoxide anion radical and nitric oxide at the same time. In addition, the present study tests whether or not the components directly scavenge peroxynitrite itself. The results showed that the components with the aromatic ortho-dihydroxyl groups (catechol) were more potent inhibitors of peroxynitrite formation by SIN-1. In particular, the methyl ester form of chlorogenic acid showed the most potent inhibition. At $5\;{\mu}M$ concentration, the order of minimizing peroxynitrite formation were : methyl chlorogenic acid > quercitrin > quercetin 3-O-sambubioside > chlorogenic acid > afzelin. Authentic peroxynitrite was directly scavenged by the components in a manner similar to peroxynitrite formation by SIN-1. In particular, when compared with penicillamine as a positive control, methyl chlorogenate was as effective in inhibiting peroxynitrite formation and approximately 2 times more effective in scavenging an authentic peroxynitrite. These results demonstrate therefore, that components extracted from the leaves of Eriobotrya japonica effectively scavenged peroxynitrite.

• The Journal of Korean Medicine
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• v.21 no.4
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• pp.183-192
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• 2000

Objectives : The present study was carried out to investigate the effects of Danchun-hwan(DCH) on the peroxynitrite-induced neural cell death in human neuroblastoma cell line, SH-SY5Y. Methods : The cultured cells were pretreated with DCH and exposed to 3-morpholinosydnonimine(SIN-1) that simultaneously generates NO and superoxide, thus possibly forming peroxynitrite. The cell damage was assessed by using MTT assay and crystal violet staining. Results : Exposure of the cells to SIN-1 for 24hr induced 75% apoptotic cell death, as evaluated by the occurrence of morphological nuclear changes characteristic of apoptosis using 4', 6-diamidino-2-phenylinole(DAPI). However, pretreatment of SH-SY5Y with the water extracts of DCH, inhibited the apoptotic cell death in a dose-dependent manner. DCH also inhibited SIN-1-induced apoptotic caspase 3-like protease activity in a dose-dependent manner. DCH recovered the depleted glutathione levels by SIN-1. Conclusions : Taken together, it is suggested that DCH protected human neuroblastoma cell line, SH-SY5Y, from the free radical injury mediated by peroxynitrite by a mechanism of elevating antioxidant, GSH.

• Korean Journal of Agricultural Science
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• v.42 no.4
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• pp.407-414
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• 2015

To increase antioxidative activity of Chungkukjang, the protective effect of Seoritae Chungkukjang (SC) added with green tea powder against oxidative stress was evaluated under the cellular system using LLC-$PK_1$ cells. The treatment of 3-morpholinosydnonimine showed increase in lipid peroxidation, and decrease in endogenous anti-oxidant enzymes activity and cell viability. The methanol extract of SC inhibited lipid peroxidation by 70.9%, and significantly increased cell viability up to more than 33.2%. In addition, it enhanced superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities. Particularly, the addition of green tea in SC exerted protective effect against oxidative stress by ONOO- through elevation in activities of SOD and GSH-Px, and inhibition of lipid peroxidation. More addition of green tea showed stronger protective activity. These results suggest that the addition of green tea to SC leads to the increase in the antioxidative effect of Chungkukjang through elevation in antioxidative enzyme activities and protection from lipid peroxidation.

• Ocean and Polar Research
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• v.29 no.2
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• pp.187-191
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• 2007

In this study, antioxidant activities of acetone/dichloromethane and methanol extracts of Arctic seaweeds were investigated. The antioxidant properties of both extracts of arctic seaweed were evaluated using two different peroxynitrite tests, including scavenging power on authentic peroxynitrite and inhibitory activities on peroxynitrite generation from 3-morpholinosydnonimine (SIN-1) producing superoxide anion and nitric oxide simultaneously in vitro. At concentration of $10\;{\mu}g/ml$, the acetone/dichloromethane and methanol extracts of Odonthalia dentata exhibited 54.6 and 64.2% inhibition against peroxynitrite generation from SIN-1 while they exhibited 24.3 and 23.0% scavenging activities on authentic peroxynitrite, respectively. On the other hand, the acetone/dichloromethane extract of Polysiphonia stricta exhibited 61.8% inhibition only against the induced peroxynitrite from SIN-1. Furthermore, the crude extracts of Odonthalia dentata and Polysiphonia stricta were fractionated into n-hexane, 85% aq. MeOH, n-BuOH, and $H_2O$ fractions, successively, and only 85% aq. MeOH fraction exhibited the best inhibition.

• Archives of Pharmacal Research
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• v.28 no.7
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• pp.810-815
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• 2005

Previously, we have shown that astrocytes deprived of glucose became highly vulnerable to peroxynitrite, and adenosine and its metabolites attenuated the gliotoxicity via the preservation of cellular ATP level. Here, we found that adenosine and related metabolites prevented the disruption of mitochondrial transmembrane potential (MTP) in glucose-deprived rat primary astrocytes exposed to 3-morpholinosydnonimine (SIN-1), a peroxynitrite releasing agent. Exposure to glucose deprivation and SIN-1(2h) significantly disrupted MTP in astrocytes, and adenosine prevented it in dose-dependent manner with an $EC_{50}\;of\;5.08{\mu}M$. Adenosine also partially prevented the cell death by myxothiazol, a well-known inhibitor of mitochondrial respiration. Blockade of adenosine deamination or intracellular transport with erythro-9-(-hydroxy-3-nonyl)adenosine (EHNA) or S-(4-nitrobenzyl)-6-thioinosine (NBTI), respectively, completely reversed the protective effect of adenosine. Other purine nucleos(t)ides including inosine, guanosine, ATP, ADP, AMP, ITP, and GTP also showed similar protective effects. This study indicates that adenosine and related purine nucleos(t)ides may protect astrocytes from peroxynitrite-induced mitochondrial dysfunction.

• Ocean and Polar Research
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• v.33 no.2
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• pp.185-191
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• 2011

Crude extracts of Limonium tetragonum and their solvent-partitioned fractions were evaluated for their potential to scavenge authentic $ONOO^-$, and $ONOO^-$ derived from 3-morpholinosydnonimine (SIN-1). Four flavonol glycosides (1-4) were isolated by activity-guided separation. Their chemical structures were elucidated by extensive 2 D NMR experiments and by comparison with published spectral data. These compounds were also estimated for their peroxynitrite scavenging effects. The scavenging ratios of compounds 1-4 on authentic $ONOO^-$ were 56, 37, 56, and 54%, respectively, at a concentration of 1 ${\mu}M$. On the other hand, the inbihition ratios of compounds 1-4 against $ONOO^-$ generation from SIN-1 were 59, 39, 44, and 54% at the same concentration, respectively.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.20 no.8
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• pp.303-312
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• 2019

Oxidative stress in kidneys can precede the development of chronic renal injury. We investigated the antioxidative effect of membrane-free stem cell extract (MFSCE) from adipose tissue in LLC-$PK_1$ renal proximal tubule cells. Treatment of LLC-$PK_1$ cells with MFSCE showed the up-regulation of heme-oxygenase-1, thioredoxin reductase 1, and NADPH quinine oxidoreductase-1 protein expressions, which are proteins related with antioxidative activities. When oxidative stress was induced by 3-morpholinosydnonimine (SIN-1), cell viability was decreased, indicating that LLC-$PK_1$ cells were damaged by SIN-1. However, MFSCE significantly elevated cell viability from 58.84% to 64.43% at the concentration of $2.5{\mu}g/mL$ in oxidative stress-induced LLC-$PK_1$ cells. Furthermore, MFSCE ameliorated inflammation and apoptosis in SIN-1-treated LLC-$PK_1$ cells by modulating protein expressions. Inducible nitric oxide synthase and cyclooxygenase-2 protein expressions were down-regulated when LLC-$PK_1$ cells were treated with MFSCE. Apoptosis-related proteins, including B-cell lymphoma-2-associated X protein/B-cell lymphoma-2 ratio, cleaved caspase-3, and cleaved-poly (ADP-ribose) polymerase, were also down-regulated. It indicated that MFSCE protected apoptosis against oxidative stress in LLC-$PK_1$ cells. Taken together, these results suggested that MFSCE had a protective effect against SIN-1-induced oxidative stress in LLC-$PK_1$ cells. Therefore, MFSCE could be a promising therapeutic agent for oxidative stress-induced renal injury.

• Journal of Applied Biological Chemistry
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• v.57 no.3
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• pp.205-210
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• 2014

Persicaria orientalis (L.) Spach (Po) and Potentilla fragarioides var. major Maxim (Pf) extracts were analyzed to investigate anti-inflammation through their suppressing effects on free radicals such as reactive oxygen species (ROS). In addition, with regard to Po and Pf, an analysis was conducted of their inhibitory effect on nitric oxide, which is produced in lipopolysaccharide (LPS)-treated murine macrophage RAW 264.7 cells, and their inhibitory effect on the translocation of the nucleus of nuclear factor-kappa B (NF-${\kappa}B$). The $IC_{50}$ value of ROS, which was induced by $50{\mu}M$ 3-morpholinosydnonimine hydrochloride (SIN-1), was found to be $23.35{\pm}1.27{\mu}g/mL$ due to the effect of the Po extract, and $8.46{\pm}1.22{\mu}g/mL$ due to the effect of the Pf extract. In addition, the $IC_{50}$ value of peroxynitrite treated with the Po extract was $2.19{\pm}0.04{\mu}g/mL$, whereas that of peroxynitrite treated with the Pf extract was $0.80{\pm}0.02{\mu}g/mL$. ROS and peroxynitrite were induced by $50{\mu}M$ 3-morpholinosydnonimine hydrochloride. There was an increase in the amount of nitric oxide in the RAW 264.7 cells treated with LPS ($1{\mu}g/mL$), whereas the level of NO was observed to significantly and dose-dependently decrease in the cells treated with Po and Pf. The amount of nitric oxide produced by the group treated with $10{\mu}g/mL$ of the Pf extract was $11.45{\pm}0.57{\mu}M$. Furthermore, the Po extracts inhibited the translocation of the nucleus of NF-${\kappa}B$ in LPS-treated RAW 264.7 cells. Therefore, it is highly possible that Po and Pf have anti-inflammatory properties.