• Bulletin of the Korean Chemical Society
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• 제28권12호
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• pp.2329-2332
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• 2007

Salsolinol, endogenous neurotoxin, is known to be involved in the pathogenesis of Parkinson's disease (PD). In the present study, we have investigated the oxidative damage of DNA induced by the reaction of salsolinol with Cu,Zn-SOD. When plasmid DNA incubated with salsolinol and Cu,Zn-SOD, DNA cleavage was proportional to the concentrations of salsolinol and Cu,Zn-SOD. The salsolinol/Cu,Zn-SOD system-mediated DNA cleavage was significantly inhibited by radical scavengers such as mannitol, ethanol and thiourea. These results indicated that free radicals might participate in DNA cleavage by the salsolinol/Cu,Zn-SOD system. Spectrophotometric study using a thiobarbituric acid showed that hydroxyl radical formation was proportional to the concentration of salsolinol and was inhibited by radical scavengers. These results indicated that hydroxyl radical generated in the reaction of salsolinol with Cu,Zn-SOD was implicated in the DNA cleavage. Catalase and copper chelators inhibited DNA cleavage and the production of hydroxyl radicals. These results suggest that DNA cleavage is mediated in the reaction of salsolinol with Cu,Zn-SOD via the generation of hydroxyl radical by a combination of the oxidation reaction of salsolinol and Fenton-like reaction of free copper ions released from oxidatively damaged SOD.

• BMB Reports
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• 제40권5호
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• pp.684-689
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• 2007

The endogenous neurotoxin, 1-methyl-6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline (salsolinol), has been considered a potential causative factor for the pathogenesis of Parkinson's disease (PD). In the present study, we examined the pattern of human Cu,Zn-superoxide dismutase (SOD) modification elicited by salsolinol. When Cu,Zn-SOD was incubated with salsolinol, some protein fragmentation and some higher molecular weight aggregates were occurred. Salsolinol led to inactivation of Cu,Zn-SOD in a concentration-dependent manner. Free radical scavengers and catalase inhibited the salsolinol-mediated Cu,Zn-SOD modificaiton. Exposure of Cu,Zn-SOD to salsolinol led also to the generation of protein carbonyl compounds. The deoxyribose assay showed that hydroxyl radicals were generated during the oxidation of salsolinol in the presence of Cu,Zn-SOD. Therefore, the results indicate that free radical may play a role in the modification and inactivation of Cu,Zn-SOD by salsolinol.

• BMB Reports
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• 제37권3호
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• pp.325-329
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• 2004

Oxidation of catecholamines may contribute to the pathogenesis of Parkinson's disease (PD). The effect of the oxidized products of catecholamines on the modification of Cu,Zn-superoxide dismutase (SOD) was investigated. When Cu,Zn-SOD was incubated with the oxidized 3,4-dihydroxyphenylalanine (DOPA) or dopamine, the protein was induced to be aggregated. The deoxyribose assay showed that hydroxyl radicals were generated during the oxidation of catecholamines in the presence of copper ion. Radical scavengers, azide, N-acetylcysteine, and catalase inhibited the oxidized catecholamine-mediated Cu,Zn-SOD aggregation. Therefore, the results indicate that free radicals may play a role in the aggregation of Cu,Zn-SOD. When Cu,Zn-SOD that had been exposed to catecholamines was subsequently analyzed by an amino acid analysis, the glycine and histidine residues were particularly sensitive. These results suggest that the modification of Cu,Zn-SOD by oxidized catecholamines might induce the perturbation of cellular antioxidant systems and led to a deleterious cell condition.

• BMB Reports
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• 제31권5호
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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.

• 대한한의학회지
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• 제26권3호
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• pp.204-214
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• 2005

Objectives : The verity extract of the Gagam-Hyungbang-Gihwang-tang (GHG) was assessed to determine the mechanisms of its antioxidant activity. Methods : The fellowing effects were measured : GHG exhibited a concentration-treatment; scavenging ${\alpha},\;{\alpha}-diphenyl-\beta-picrylhydrazyl$ (DPPH) radical, linoleic acid oxidation in a thiocyanate assay system, and superoxide anion, hydroxyl radical-induced DNA nicking. We investigated mRNA levels such as superoxide-dismutase. Results : The GHG extract showed dose-dependent free radical scavenging activity, including DPPH radicals, hydroxyl radicals, and superoxide anion, using different systems. The GHG was also found to be effective in protecting plasmid DNA against the strand breakage induced by hydroxyl radicals in Fenton's reaction mixture. Furthermore, SOD-1 mRNA expression levels increased in tat hepatoma H4IIE cells Conclusions : We expect that GHG will to helpful to the development of antioxidant activity treatments.

• 생명과학회지
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• 제7권4호
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• pp.361-370
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• 1997

Sprague-Dawley male rats were fed experimental diet, and also were orally daministerred with 3.0% ultra low viscosity (ULV) sodium alginate-added functional drink(AL-3.0% group : HAEJOMIN), 5.0% polyedxtrose(PD)-added drink(PD-5.0group) and 2.5% polydextrose-added drink(PD-2.5 group) for 8 weeks. Effect of rhese dietary fiber-added functional drinks on body weight, feed and gross efficincies, triglyceride and cholesterol levels, LDL- cholesterol levels, hydroxyl radical and malondialdehyde levels, and superoxide dismutase (SOD) activity in serum of SD rats were evaluated. Administration of AL-3.0 drink and PD-5.0 drink resulted in a marked inhibition in increase of body weight compared with control and PD-2.5 groups for 8 weeks. Inhibition effect in body weight in 3.0% alginic acid-added drink )AL-3.0 froup_ showed a same trend in 5.0% polydextrose(PD)-added drink (PD-5.0 group)(p<0.001). Therefore, it is found that inhibition effects of obesity in 3.0% alginic acid-added drink were higher 2 times than that in same concentration of polydextrose(PD)-added drink. Triglyceride and cholesterol levels in AL-3.0 and PD-5.0 groups significantly decreased to 25$\sim$30% compared with control group(p<0.01$\sim$0.001), but there were no significant differences in these drinks. LDL-cholesterol levels in AL-3.0 group significantly decreased about 15% compared with PD-5.0 group, but atherogenic index in AL-3, 0 group showed a similar trend to that in PD-5.0 group. Hydroxyl radical formations and lipid peroxide(LPO) levels in AL-3, 0 and PD-5.0 groups significantly decreased to 15% and 20%, respectively, compared with control group(p, 0.05$\sim$0.01), but there were no significant differences in these drinks. Superoxide, dismutase(SOD) activity in AL-3.0 group significantly higher (about 255) than those in control and PD-5.0 groups(p<0.01). These results suggest that administration of ULV-sodium alginate-added functional drink(HAEJOMIIN) effectively can not only inhibit obesity, but also can intervent chronic degenerative disease and aging process.

• 생명과학회지
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• 제9권3호
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• pp.276-281
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• 1999

해양오염의 진단을 위한 생화학적 오염지표 설정의 기초연구의 일환으로, 오염이 심각한 남해산 홍합(Mytilus coruscus)의 근육중의 활성산소종 및 그들의 제거효소의 활성을 $분석{\times}평가하였다.$ 이들 홍합의 근육중의 단백질의 향량은 남해안 양식산 홍합의 근육중의 단백질 함량이 동해안의 포항의 자연산 홍합(대조군)의 단백질 함량에 비해 4~14%나 유의적으로 감소하였다.(p<0.05-0.01).남해안의 양식산 홍합의 근육중의 수퍼옥시드 라디칼의 생성량은 대조군으로 사용한 동해안 포항의 자연산 홍합의 근육중의 수퍼옥시드 라디칼의 생성량 대비 82~138%정도나 높앗다. 또한 남해안의 양식산 홍합의 근육중의 히드록시 라디칼의 생성량은 대조군으로 사용한 동해안 포항의 자연산 홍합의 근육중의 히드록시 라디칼의 생서량 대비 9~25%정도나 높았다.남해안의 양식산 홍합의 근육중의 SOD의 활성은 동해안 포항의 자연산 홍합의 근육중의 SOD의 활성대비 16~28%정도나 낮았다. 이상의 영구결과에서 볼 때 남해안 홍합의오염 지표로서는 단백질 함량의 감소, 활성산소종의 증가 및 활성산소의 제거효소로서 SOD의 활성감소로 평가각 가능하다라는 사실을 확인할 수 있었다.

• 생명과학회지
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• 제10권5호
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• pp.504-510
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• 2000

This study was designed to investigate the effects of mulberry (Morus alba L.) leaf extract (MLE) on oxygen radicals and their scavenger enzymes in liver membranes of rats. Sprague-Dawley (SD) male rats (160$\pm$10 g) were fed basic diet (control group), and experimental diets (MLE-100 and MLE-300 groups) added 100 and 300 mg/kg BW/day for 6 weeks. Hydroxyl radical (.OH) levels resulted in a significant decreases (15.2% and 18.1%, 5.6% and 8.0%, respectively) in liver mitochondria and microsomes could be not obtained. These are no significant differences in superoxide radical ($O_2$) levels of liver cytosol in MLE-100 and MLE-300 groups compared with control group. Lipid peroxide (LPO) levels were slightly decreased about 13.6% and 6.1% in liver mitochondria and microsomes of MLE-300 group compared with control group. Oxidized protein (OP) levels were remarkably decreased about 16.9% and 27.2% in liver microsomes only of MLE-100 and MLE-300 group compared with control group. Mn-SOD activities in liver mitochondria were remarkably increased (18.2% and 28.7%, respectively) in MLE-100 and MLE-300 groups, and Cu,Zn-SOD activities in liver cytosol were also significantly increased (11.3% and 20.2%, respectively) in MLE-100 and MLE-300 groups compared with control group. Mn-SOD activities in liver mitochondria were remarkably increased (18.2% and 28.7%, respectively) in MLE-100 and MLE-300 groups, and Cu,Zn-SOD activities in liver cytosol were also significantly increased (11.3% and 20.2%, respectively) in MLE-100 and MLE-300 groups compared with control group, but significant difference between GSHPx activities in liver cytosol could be not obtained. These results suggest that anti-aging effect of mulberry leaf extract (MLE) may play a pivotal role in attenuating a various age-related changes.

• 한국잠사곤충학회지
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• 제42권2호
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• pp.93-98
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• 2000

This study was designed to investigate the effect of silkworm powder on oxygen radicals and their scavenger enzymes in brain membrances of SD rats. Hydroxyl radical (OH) levels resulted in a considerable decreases in brain mitochondria fraction. Superoxide radical (O$_2$) levels were a slightly decreased in brain cytosol fraction. Lipid peroxide (LPO) and Oxidized protein (OP) levels were significantly decreased in brain mitochondria and microsomes fraction. Mn-superoxide dismutase (SOD) activity was remarkably increased in the mitochondria fraction. Cu and Zn-SOD activities were effectively increased in brain cytosol fraction. GSHPx activity was considerably increased in brain cytosol fraction. These results suggest that anti-aging effect of silkworm plays an effective role in attenuating an oxidative stress and increasing a scravenger enzyme activity in brain membranes.

• 생명과학회지
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• 제23권12호
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• pp.1428-1435
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• 2013

본 연구의 목적은 melanocyte (B16F1)에서 quercetin과 kaempferol을 포함하는 미나리 에탄올 추출물(OJE)의 멜라닌 합성효과에 미치는 영향을 조사한 것이다. OJE가 세포수준에서 멜라닌 합성을 억제하는지를 조사하기 위하여 여러 농도의 OJE 존재 하에서 B16F1세포를 배양하였다. 현재 연구에서 DPPH radical scavenging, reducing power, lipid peroxidation 및 DNA oxidation에 미치는 항산화 효과는 cell free system에서 평가되었다. 더욱이 멜라닌 생성에 대한 OJE 효과는 dopaquinone (DOPA) assay 및 tyrosinase 활성으로 결정되었다. 뿐만 아니라 superoxide dismutase (SOD)-1, -2, glutathione reductase (GSH)와 같은 항산화 효소 및 tyrosinase의 단백질발현이 western blot 분석을 이용하여 평가되었다. 본 연구에서 OJE는 지질과산화 억제효과를 나타내었고 fenton 반응에 의해서 생성되는 hydroxyl radical에 의하여 유발되는 DNA 산화를 보호하였다. OJE는 50 ${\mu}g/ml$ 이상에서 멜라닌 합성을 증가시켰고 tyrosinase 활성도 50 ${\mu}g/ml$에서 검출되었다. Western blot 분석에서는 OJE가 농도에 비례하여 tyrosinase SOD-1, -2 및 GSH의 발현 수준을 증가시켰다. 이러한 발견들은 항산화 효과를 가진 OJE가 melanocyte에서 tyrosinase 활성과 melanin 생성을 조절할 수 있어 피부를 산화스트레스로부터 보호할 수 있다는 것을 암시하고 있다.