• Bulletin of the Korean Chemical Society
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• v.32 no.3
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• pp.964-972
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• 2011

This work investigated the difference between $Fe^{2+}$ autoxidation-induced and Fenton-type cleavage of pBR322 plasmid DNA. $^{\cdot}OH$ generation reactions in the absence and presence of $H_2O_2$ under various conditions were also investigated. Although both the $Fe^{2+}$ autoxidation and Fenton-type reactions showed DNA cleavage and $^{\cdot}OH$ generation, there were significant differences in their efficiencies and reaction rates. The rate and efficiency of the cleavage reaction were higher in the absence of 1.0 mM of $H_2O_2$ than in its presence in 20 mM phosphate buffer. In contrast, the $^{\cdot}OH$ generation reaction was more prominent in the presence of $H_2O_2$ and showed a pH-independent, fast initial reaction rate, but the rate was decreased in the absence of $H_2O_2$ at across the entire tested pH range. Studies using radical scavengers on DNA cleavage and $^{\cdot}OH$ generation reactions in both the absence and presence of $H_2O_2$ confirmed that both reactions spontaneously involved the active oxygen species $^{\cdot}OH$, ${O_2}^{\cdot-}$, $^1O_2$ and $H_2O_2$, indicating that a similar process may participate in both reactions. Based on the above observations, a new mechanism for the $Fe^{2+}$ autoxidation-induced DNA cleavage reaction is proposed.

• Applied Biological Chemistry
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• v.43 no.2
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• pp.141-147
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• 2000

Interrelation between the antioxidative activity of hot-water extracts of 130 medicinal plants and their cellular antimutagenic activity was investigated. Antioxidative activity was evaluated by assaying electron-donation to DPPH free radical and scavenging of hydroxyl radical $({\cdot}OH)$ generated through Fenton rection, respectively. All medicinal plants examined in this study exhibited markedly electron-donating ability and radical scavenging ability in each assay system. The results demonstrated the fact that Pilbal (Piper longum L.) is the strongest in electron-donating activity, on the other hand, that Seokgok (Dendrobium moniliforme L.) is the strongest in ${\cdot}OH$ scavenging activity. When evaluated their antioxidative activities, 24 medicinal plants including Jimo (Anemarrhena asphodeloides Bunge) were found to be the medicinal plants carrying strong antioxidative activity, which exhibited more than 50% activity compared to the control group in both electron-donating and free radical scavenging. The experiment was also performed to examine whether 11 medicinal plants having significant antimutagenicity damage DNA in the presence of $Cu^{2+}$, showing the fact that all samples tested, except Taeksa (Alisma canaliculatum All. Br.), Paekjain (Nitraria sibirica Pall) and Ohyak (Lindera strychifolia Sieb. et Zucc. Villar) are capable of inducing DNA strand break. We also found that Taeksa and Paekjain strongly block DNA strand break induced by chemical mutagen mitomycin C.

• Journal of Life Science
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• v.21 no.1
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• pp.56-61
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• 2011

This study was performed to investigate effective extract conditions in fronds of the Ostrich fern (Matteuccia struthiopteris) to increase antioxidant compound contents and antioxidant capacity. Powder (1 g) of lyophilizated fronds were mixed with 3 kinds of solvents (MeOH, 80% EtOH and water). Extractions were carried out using not only immersion (room temp.), heating ($60^{\circ}C$) and stirring (200rpm) for 6 hr, but also through sonication in a 42 kHz ultrasonic bath for 15, 30 and 45 min. Extracts were filtrated and measured for contents of soluble solids (SS), total polyphenols (TP; tannic acid as a standard) and total flavonoids (TF; Naringin as a standard). Antioxidant activity was expressed as $RC_{50}$ for DPPH and ABTS radical scavenging. SS (0.317 $g{\cdot}g^{-1}$ db), TP (70.90 $mg{\cdot}g^{-1}$ db) and TF (41.53 $mg{\cdot}g^{-1}$ db) contents reached their highest levels when 30 minute sonication extraction with 80% EtOH was performed, and the highest DPPH and ABTS scavenging activity was observed in the same extraction conditions ($RC_{50}$=0.14 $mg{\cdot}ml^{-1}$ and 0.09 $mg{\cdot}ml^{-1}$, respectively). From the present investigation, it can be concluded that fronds of the ostrich fern can be used as a natural material for antioxidants, and sonication for 15-30 min with 80% EtOH is an ideal extraction method for increasing their antioxidant effects and saving extraction time.

• Preventive Nutrition and Food Science
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• v.17 no.4
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• pp.261-268
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• 2012

Ribes diacanthum Pall (RDP) is a member of the Saxifragaceae family. The plant is traditionally used in Mongolia for the treatment of various ailments associated with kidney and bladder's diseases, cystitis, kidney stone, and edema. This study was aimed to investigate antioxidant activities of different solvent extracts of whole Pall plants, based on ferric-reducing antioxidant potential (FRAP), 2,2'-azinobis(3-ethybenzothiazoline-6-sulfonic acid) ($ABTS{\cdot}+$) radical scavenging activity, 1,1-diphenyl-2-picrydrazyl ($DPPH{\cdot}$), and hydroxyl (${\cdot}OH$) radical scavenging activities. Additionally, total flavonoids and phenolic contents (TPC) were also determined. The ethyl acetate extract of RDP (EARDP) had a remarkable radical scavenging capacity with an $IC_{50}$ value of 0.1482 mg/mL. In addition, EARDP was shown to be higher in total phenolic and flavonoid contents than the methanol extract of RDP (MRDP). Moreover, the EARDP had the predominant antioxidant capacity, DPPH, hydroxyl, and ABTS radical scavenging activities and ferric reducing power. These results suggest a potential for R. diacanthum Pall extract as a functional medicinal material against free-radical-associated oxidative damage.

• The Korean Journal of Pharmacology
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• v.21 no.1
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• pp.12-26
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• 1985

The generation of $O^{-}_{2}{\cdot}$ and its toxic effects were studied with rat brain mitochondria. The production of $O^{-}_{2}{\cdot}$ from mitochondria in the presence of succinate and antimycin was demonstrated by SOD-inhibitable reduction of NBT. Although succinate can support the $O^{-}_{2}{\cdot}$ formation, the highest rate needs antimycin indicating that blockade of electron flow in the respiratory chain augments the univalent reduction of molecular oxygen. Under this condition, $H_2O_2$ was also observed to be produced. But its formation appears to be derived from the dismutation of the primary product, $O^{-}_{2}{\cdot}$ since the rate of $H_2O_2$ production was markedly decreased by NBT and ferricytochrome c. The $O^{-}_{2}{\cdot}$ and $H_2O_2$ produced were able to cause toxic actions to mitochondrial and extra-mitochondrial components as shown by lipid peroxidation of mitochondrial membrane, and inactivation and lysis of isocitrate dehydrogenase and erythrocytes added to the medium, respectively. In all the toxic actions observed, $Fe^{++}$ was required. It appears that in the toxic actions $OH{\cdot}$ generated from the iron-catalyzed Haber-Weiss reaction acts as a mediator. This was supported by the finding that mitochondria in the presence of succinate and antimycin produced ethylene from methional, and $Fe^{++}$ added increased the ethylene production. The observed toxic actions of mitochondrial $O^{-}_{2}{\cdot}$ may provide evidence supporting a potential role of mitochondria as a source of oxygen radicals to cause tissue damage.

• Journal of Applied Biological Chemistry
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• v.50 no.3
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• pp.115-119
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• 2007

To discover the sources with antioxidant and tyrosinase inhibitory activities in Korean traditional medicines, 10 extract of medicinal plants were screened for their potential to scavenge stable 1,1-diphenyl-2-picryhydrazyl (DPPH) free radical, inhibit hydroxyl radical (${\cdot}OH$), total phenolic content, and inhibition of tyrosinase. The potency of DPPH radical scavenging activity was shown in the extract of Ulmus davidiana var. japonica Nakai that has a greater effect with $IC_{50}$ values of $6.49{\pm}5.43{\mu}g/mL$, than BHA ($IC_{50}=20.99{\pm}0.74{\mu}g/mL$), L-ascorbic acid $(IC_{50}=20.59{\pm}1.06{\mu}g/mL),\;and\;{\alpha}-tocopherol\;(IC_{50}=25.55{\pm}0.26{\mu}g/mL)$ as a positive control. The ${\cdot}OH$ scavenging activities were observed in the plants tested. Acanthopanax senticosus, Cirsium setiders, U. davidiana exhibited scavenging activity of more than 60% at $500{\mu}g/mL$. The scavenging activity(%) of BHA and a-tocopherol were 64.32 and 55.87% at $500{\mu}g/mL$, respectively. The total phenolic content was determined, in order to assess its effect on the extract's antioxidant activity. The total phenoic content of $33.37{\pm}0.52mg/g$ was conformed by methanolic extract of U. davidiana. The U. davidiana and Morus bombycis exhibited tyrosinase inhibitory activity with a $34.28{\pm}1.32\;and\;75.57{\pm}1.10%$, respectively. In particular, M. bombycis has stronger tyrosinase inhibitory activity than albutin with $36.48{\pm}3.56%$ as a positive control. This work showed that the inhibitory abilities of Korean medicinal plants, such as U. davidiana and M. bombycis, on DPPH free radical, inhibit hydroxyl radical (${\cdot}OH$), and inhibition of tyrosinase and total phenolic content, can be useful in the prevention and treatment of free radical-relate disease. Investigations into further isolation of inhibitory principles of U. davidiana and M. bombycis are now in progress.

• Korean Journal of Agricultural Science
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• v.45 no.4
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• pp.761-767
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• 2018

Degenerative diseases are commonly associated with excess free radicals. Acer okamotoanum, a plant endemic to Korea, is reported to have anti-oxidant, anti-cancer, and anti-viral activities. We previously isolated flavonoids from the ethyl acetate fraction of A. okamotoanum such as quercitrin (QU), isoquercitrin (IQ), and afzelin (AF). In the present study, the in vitro antioxidant activity of flavonoids such as QU, IQ, and AF isolated from the ethyl acetate fraction of A. okamotoanum were investigated by measuring the free radical scavenging activity including 1,1-diphenyl-2-picrylhydrazyl (DPPH), hydroxyl radical ($^{\cdot}OH$), and superoxide anion ($O_2{^-}$). The flavonoids (QU, IQ, and AF) concentration-dependently showed a DPPH radical scavenging activity. In particular, QU and IQ showed a higher DPPH radical scavenging activity than that of AF. In addition, the flavonoids (QU, IQ, and AF) at $10{\mu}g/mL$ showed over an 80% scavenging effect against $^{\cdot}OH$ radical production. Furthermore, the $O_2{^-}$ radical scavenging activity of the flavonoids, QU, IQ, and AF increased in a dose-dependent manner. Particularly, IQ exerted the strongest scavenging activities against $^{\cdot}OH$ and $O_2{^-}$ radicals among the other flavonoids. These results indicate that the flavonoids from A. okamotoanum, in particular IQ, would have a protective activity against oxidative stress induced by free radicals, and potentially be considered as a natural antioxidant agent.

• Reproductive and Developmental Biology
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• v.35 no.2
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• pp.191-195
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• 2011

The present study was conducted to examine the reactive oxygen species (ROS) generation levels in porcine parthenogenetic embryos. Porcine in vitro matured oocytes were activated by the combination of electric stimulus and 6-DMAP before in vitro culture. Porcine oocytes and parthenogenetic embryos were stained in 10 ${\mu}M$ dichlorohydrofluorescein diacetate (DCF) or 10 ${\mu}M$ hydroxyphenyl fluorescein (HPF) dye each for 30 min at $39^{\circ}C$. The fluorescent emissions from the samples were recoded as JPEG file and the intensity of fluorescence in oocytes and embryos were analyzed. $H_2O_2$ and $^{\cdot}OH$ radical levels of porcine oocytes were reduced immediately after electric stimulation. However, $H_2O_2$ and $^{\cdot}OH$ radical levels of parthenogenetic embryos were increased with time elapsed after electric stimulation from 0 h to 3 h and after DMAP culture. During in vitro culture, $H_2O_2$ and $^{\cdot}OH$ radical levels were gradually increased from the one-cell stage to the two- and four-cell stages. The result of the present study suggests that the ROS was not increased by electric pulse in porcine embryos. Rather than it seems to be associated with the stage of development and the culture condition.

• Preventive Nutrition and Food Science
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• v.16 no.3
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• pp.224-229
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• 2011

The radical scavenging activity and inhibition effect from lipid peroxidation induced by peroxyl radical of methanol extract from Perilla frutescens and its active compound, rosmarinic acid (RA), were investigated in vitro. The treatment of extract and RA scavenged 1,1-diphenyl-2-picrylhydrazyl, hydroxyl radical (${\cdot}OH$) and nitric oxide in a concentration-dependent manner. In particular, the extract and RA showed strong radical scavenging activity against ${\cdot}OH$, the most toxic and reactive radical. In addition, Perilla frutescens and RA effectively inhibited lipid oxidation induced by sodium nitroprusside and 2,2'-azobis(2-aminopropane) dihydrochloride, determined by the ferric thiocyanate method. The present results suggest that Perilla frutescens and RA play a protective role against oxidative stress induced by free radical and lipid peroxidation.

• BMB Reports
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• v.31 no.5
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• pp.492-497
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• 1998

Cu,Zn-superoxide dismutase (SOD) was purified from horseradish by using Mono Q and Superose 12 FPLC column chromatography. The native molecular mass of the purified enzyme was approximately 33 kDa, as determined by gel filtration. The subunit molecular weight, as estimated by SDS-PAGE, was 16 kDa. These results indicated that the native enzyme is a homodimer. We investigated the free radical-generating function of horseradish Cu,Zn-SOD by using a chromogen, 2,2'-azinobis-(3-ethylbenzthiazoline-6-sulfonate) (ABTS) which reacts with ${\cdot}OH$ radicals to form $ABTS^{+{\cdot}}$ The formation of $ABTS^{+{\cdot}}$ was required for both active Cu, Zn-SOD and $H_2O_2$. The optimal pH for the free radical-generating activity of this enzyme was 6.0-8.0, and it retained about $40^{\circ}C$ of its maximum activity when exposed at $40^{\circ}C$ for 15 min. A neutral scavenger, ethanol, inhibited the $ABTS^{+{\cdot}}$ formation by horseradish Cu, Zn-SOD more effectively than that by the mammalian enzyme. These results suggest that the active channel of horseradish enzyme is slightly larger than that of the mammalian enzyme.