• Proceedings of the Plant Resources Society of Korea Conference
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• 2011.10a
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• pp.19-19
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• 2011

예비실험 중 한국의 울릉도에 자생하고 취나물로 이용되고 있는 섬쑥부쟁이(Aster glehni Franchet et Sckmidt, Compositae)가 페놀성 화합물 함량이 높고 peroxynitrite 소거효과가 높았기 때문에 이를 이용한 추출물의 제조를 통한 건강기능식품을 창출하고자 하였다. 건강식품을 위한 추출물 제조를 위하여 물과 에탄올만을 이용하여 페놀성 화합물 고함유 추출물의 제조가 가능하고 높은 peroxynitrite 소거효과를 나타내는 추출물을 결정하고자 하였다. 즉, 추출용매로 Water-EtOH 비율을 조절하여 추출하여 추출물의 함량, HPLC를 이용한 caffeoylquinic acid(CQ) 화합물의 함량분석 및 peroxynitrite 소거효과를 비교하였다. Water-EtOH (7:3)으로 추출한 섬쑥부쟁이 추출물은 CQ 함량이 높고, 높은 peroxynitrite 소거효과, 항경련효과, 진정효과 및 항비만효과 등이 나타났다. 건강기능식품을 위한 추출물 제조시 한국에서는 물과 에탄올만이 허용되기 때문에 CQ 함량이 높은 추출물의 제조가 가능하고 높은 peroxynitrite 소거효과를 유지할 수 있는가에 관한 실험 이외에 마우스에서 선택된 추출물의 진정효과 및 항경련효과 실험을 수행하였다. 진정효과와 항경련효과를 위하여 마우스에서 각각 pentobarbital로 유도한 수면연장효과와 PTZ로 유도한 경련에 대한 효과를 측정하여 유의성있는 활성이 확인되었으므로 이 추출물을 이용한 건강기능식품으로의 이용이 가능하다고 생각된다.

• The Korean Journal of Physiology and Pharmacology
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• v.5 no.3
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• pp.205-212
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• 2001

It has been proposed that nitirc oxide is involved in the pathogenesis of cerebral ischemia-reperfusion. Because superoxide production is also enhanced during reperfusion, the cytotoxic oxidant peroxynitrite could be formed, but it is not known if this occurs following global forebrain ischemia-reperfusion. We examined whether peroxynitrite generation is increased in the vulnerable regions after forebrain ischemia-reperfusion. Transient forebrain ischemia was produced in the conscious rat by four-vessel occlusion. Rats were subjected to 10 or 15 min of forebrain ischemia. Immunohistochemical method was used to detect 3-nitrotyrosine, a marker of peroxynitrite production. 3-Nitrotyrosine immunoreactivity was enhanced in the hippocampal CA1 area 3 days after reperfusion. Furthermore, in rats subjected to ischemia for 15 min, this change was also observed in the lateral striatal region and the lateral septal nucleus $2{\sim}3$ days after reperfusion. The cresyl violet staining of adjacent sections showed that neuronal cell death was induced in parallel with the nitrotyrosine immunoreactivity in the hippocampal CA1 area and the lateral striatal region. Our findings suggest that oxygen free radical accumulation and consequent peroxynitrite production play a role in neuronal death caused by cerebral ischemia-reperfusion.

• Toxicological Research
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• v.26 no.4
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• pp.321-327
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• 2010

The ability of 80% ethanol extracts from five medicinal plants, Aralia continentalis, Paeonia suffruticosa, Magnolia denudata, Anemarrhena asphodeloides, and Schizonepeta tenuifolia, to neutralize hydroxyl radical, peroxyl radical and peroxynitrite was examined using the total oxyradical scavenging capacity (TOSC) assay. Peroxyl radical was generated from thermal homolysis of 2,2'-azobis(2-methylpropionamidine) dihydrochloride (ABAP); hydroxyl radical by an iron-ascorbate Fenton reaction; peroxynitrite by spontaneous decomposition of 3-morpholinosydnonimine N-ethylcarbamide (SIN-1). The oxidants generated react with $\alpha$-keto-$\gamma$-methiolbutyric acid (KMBA) to yield ethylene, and the TOSC of the substances tested is quantified from their ability to inhibit ethylene formation. Extracts from P. suffruticosa, M. denudata, and S. tenuifolia were determined to be potent peroxyl radical scavenging agents with a specific TOSC (sTOSC) being at least six-fold greater than that of glutathione (GSH). These three plants also showed sTOSCs toward peroxynitrite markedly greater than sTOSC of GSH, however, only P. suffruticosa revealed a significant hydroxyl radical scavenging capacity. Seven major active constituents isolated from P. suffruticosa, quercetin, (+)-catechin, methyl gallate, gallic acid, benzoic acid, benzoyl paeoniflorin and paeoniflorin, were determined for their antioxidant potential toward peroxynitrite, peroxyl and hydroxyl radicals. Quercetin, (+)-catechin, methyl gallate, and gallic acid exhibited sTOSCs 40~85 times greater than sTOSC of GSH. These four components also showed a peroxynitrite scavenging capacity higher than at least 10-fold of GSH. For antioxidant activity against hydroxyl radical, methyl gallate was greatest followed by gallic acid and quercetin. Further studies need to be conducted to substantiate the significance of scavenging a specific oxidant in the prevention of cellular injury and disease states caused by the reactive free radical species.

• Korean Journal of Pharmacognosy
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• v.40 no.4
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• pp.376-381
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• 2009

The compositional analysis and the peroxynitrite ($ONOO^-$) scavenging assay were undertaken to demonstrate beneficial dietary effect of lettuce (Lactuca sativa L., Compositae). Quinic acid derivatives of the seven cultivars were qualitatively and quantitatively analyzed by HPLC to lead to the finding of 3,4-di-O-caffeoyquinic acid and 3-O-p-coumaroylquinic acid and their contents ranged over 2.72-4.47 mg/g and 8.97-23.26 mg/g, respectively. A cultivar Hacheong had the highest quantity of quinic acid derivatives. The peroxynitrite scavenging effect of the five cultivars (Jangsu, Gosina 27, Gopung, Yeolpungjeogchima, and Mipungjeogchugmyeon) were ranged over 7.45-8.26${\mu}g$/ml as $IC_{50}s$ while those of Hacheong and Mihong had less effect ($IC_{50}$ >10 ${\mu}g$/ml).

• KSBB Journal
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• v.19 no.1
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• pp.67-71
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• 2004

An antioxidant activity of Rosa rugosa extract and its solvent-partitioned fractions was determined not only by measuring lipid peroxide produced when a mouse liver homogenate was exposed to the air at 37$^{\circ}C$, using thiobarbituric acid (TBA) but also by evaluating the free radical scavenging effect against DPPH radical, authentic peroxynitrite, and 3-morpholinsydnonimine (SIN-1). All its partitioned fractions including crude extract showed potent scavenging effect against DPPH radical, peroxynitrite, and lipid peroxidation. n-BuOH fraction, in particular, was found to be the most effective in DPPH radical scavenging ability as well as inhibition against lipid peroxidation. The 15% aqueous MeOH fraction also showed a strong potency which was slightly lower than n-BuOH fraction. Based on these results, we suggest that Rosa rugosa could be useful for preventing an oxidative damage.

• The Journal of Korean Medicine
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• v.23 no.4
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• pp.55-63
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• 2002

Objectives: Peroxynitrite ($ONOO^{-}$), formed from the reaction of superoxide <${\cdot}O_2^{-}$) and nitric oxide (NO), is a cytotoxic species that can oxidize several cellular components such as proteins, lipids and DNA. It has been implicated in diseases such as aging process, Alzheimer's disease, rheumatoid arthritis, cancer and arteriosclerosis. Due to the lack of endogenous enzymes responsible for $ONOO^{-}$ inactivation, developing a specific $ONOO^{-}$ scavenger is of considerable importance. The aim of this study was to evaluate $ONOO^{-}$ scavenging activity and its mechanism in Cheonga-hwan (CAH). Methods: The $ONOO^{-}$ scavenging activity in CAH was assayed by measuring oxidized dihydrorhodamine 123 (DHR 123) by fluorescence. The scavenging efficacy was expressed as $IC_{50}$, showing the concentration of each sample required to cause 50% inhibition of DHR 123 oxidation. In a separate study, the protective effect of CAR on $ONOO^{-}$-induced nitration of bovine serum albumin (BSA) was investigated using immunoassay with a monoclonal anti-nitrotyrosine antibody, and a horseradish peroxidase-conjugated anti-mouse secondary antibody from sheep. Results: CAH showed potent scavenging activities of $ONOO^{-}$, NO and ${\cdot}O_2^{-}$. The data demonstrated that CAH led to decreased $ONOO^{-}$-mediated nitration of tyrosine through electron donation. CAH showed significant inhibition on nitration of bovine serum albumin by $ONOO^{-}$ in a dose-dependent manner. Conclusions: CAH can be developed as an effective peroxynitrite scavenger for the prevention of the $ONOO^{-}$ involved diseases.

• 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.

• Archives of Pharmacal Research
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• v.29 no.3
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• pp.213-217
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• 2006

The present study was undertaken to test the hypothesis that dietary antioxidants protect DNA damage induced by peroxynitrite, a potent physiological inorganic toxin. The present study showed that dietary antioxidants such as (-)-epigallocatechin gallate, quercerin, rutin, resveratrol, and ursolic acid inhibit single strand breaks in supercoiled plasmid DNA induced by 3-morpholinosydnomine N-ethylcarbamide (SIN-1), a generator of peroxynitrite through the reaction between nitric oxide and superoxide anion. The formation of 8-hydroxy-2'-deoxyguanosine (8-OH-dG) in calf thymus DNA by SIN-1 was also inhibited by dietary antioxidants. When U937 cells were incubated with 1 mM SIN-1 bolus, a significant increase of 8-OH-dG level was observed. However, oxidative DNA damage was significantly lower in the cells pre-treated with dietary antioxidants when cells were exposed to SIN-1.

• Archives of Pharmacal Research
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• v.25 no.5
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• pp.625-627
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• 2002

From the leaves of Brassica juncea, an radical scavenging isorhamnetin 7-Ο-glucoside on peroxynitrite and 1,1-diphenyl-2-picrylhydrazyl (DPPH) was isolated and characterized based on the spectroscopic evidence. The compound showed the peroxynitrite and DPPH scavenging activities with $IC_{50}$ values of 2.07$\pm$0.17 and 13.3 $\mu$M, respectively. Penicillamine and L-ascorbic acid as positive control exhibited radical scavenging activities with $IC_{50}$ values of 3.17$\pm$0.39 and 12.78 $\mu$M, respectively.

• Bulletin of the Korean Chemical Society
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• v.26 no.6
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• pp.921-924
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• 2005

It is known that metallothionein (MT) plays a role in the scavenging of free radicals, which is produced under various stress conditions. MT may function as an antioxidant that protects against oxidative damage of DNA, protein, and lipid induced by superoxide anion, hydrogen peroxide, hydroxyl radical, nitric oxide, and peroxynitrite. This study was undertaken to test the hypothesis that MT also protects from RNA damage induced by peroxynitrite, an important reactive nitrogen species that causes a diversity of pathological processes. A cell-free system was used. RNA damage was detected by the mobility of $tRNA^{Phe}$ in electrophoresis. Cleavages on tRNA were not induced by 3-morpholinosydnomine, which produces peroxynitrite directly. MT induced tRNA damage which was site specific.