• 대한수의학회지
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• 제39권5호
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• pp.878-895
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• 1999

Ischemic preconditioing (IPC), i.e., a preliminary brief episode of ischemia and reperfusion, has been shown to reduce the cell damage induced by long ischemia and reperfusion. Superoxide radical which is produced during reperfusion after ischemia was recognized as a factor of the ischemic injury and it is dismutated into $H_2O_2$ and $O_2$ by two types of intracellular superoxide dismutase (SOD), Cu,Zn-SOD in cytoplasm and Mn-SOD in mitochondria. Recently oxygen free radicals are suggested to induce the apoptosis, however mechanism of the reduced apoptosis by ischemic preconditioing was unknown, while many studies performed in mammalian heart indicated that ATP-sensitive $K^+$ ($K_{APT}$) channel activation related with the protective effects. The aim of present study is to investigate 1) whether IP upregulate the Cu,Zn-SOD and Mn-SOD activities, and 2) whether ischemic preconditioning decreases apoptosis via $K_{APT}$ channel activation in timely reperfused skeletal muscle after long ishemia. The experimental animals, Sprague-Dawley rats weighing 250~300g, were divided into 8 groups; 1) control group, 2) ischemic preconditioning only groups, 3) pinacidil, a $K_{APT}$ channel opener, treatment only groups, 4) glibenclamide, a $K_{APT}$ channel blocker, treatment only groups, 5) ischemia groups, 6) ischemia after IPC groups, 7) ischemia and pinacidil treatment groups, and 8) IP and ischemia after glibenclamide pretreatment groups. Animals of the control group were administered with the vehicle (DMSO) alone. Pinacidil (1mg/kg) was administered intravenously 5 minutes after initiation of ischemia, and glibenclamide (0.5mg/kg) was injected intravenously 20 minutes before IPC. In rats that were ischemic preconditioned, the left common iliac artery was occluded for 5 minutes followed by 5 minutes of reperfusion by three times using vascular clamp. Ischemia was done by occlusion of the same artery for 4 hours. The specimens of left rectus femoris muscle were obtained immediately (0 hour), 12 hours, 24 hours after drug administrations, IP or ischemia and reperfusion. The immunoreactivities of SOD and its alterations were observed by use of sheep antihuman Cu,Zn-SOD and Mn-SOD antibodies on the $10{\mu}m$ cryosections. The incidencies of apoptosis were observed by TUNEL methods with in situ apoptosis detection kit on $6{\mu}m$ paraffine section. The results obtained were as follows : 1. After IPC, immunoreactivities of Cu,Zn-SOD mainly in the small-sized fibers were increased by 24 hours, that of Mn-SOD at 0 hour and 24 hours. 2. No significant changes in immunoreactivities of SOD was observed in the pinacidil and in the glibenclamide treatment only groups, and in the ischemia only groups. 3. The immunoreactivities of the Cu,Zn-SOD were increased in the ischemia after IPC groups and the ischemia and pinacidil treatment groups. 4. The immunoreactivities of the Cu,Zn-SOD in the IPC and ischemia after glibenclamide pretreatment groups were not increased except for the 12 hours reperfusion group. But, Mn-SOD immunoreactivities were increased in the 0 hours, 12 hours and 24 hours after reperfusion. 5. In the control group, the IPC only groups, and the pinacidil treatment only groups, negative or trace apoptotic reactions were observed, but the positive apoptotic reaction occured in the glibenclamide treatment groups. 6. Moderate or many number of apoptosis were revealed in the ischemia groups, and also the IPC and ischemia after glibenclamide pretreatment group except for 12 hours and 24 hours after reperfusion. However, the incidence of apoptosis was decreased in the ischemia after IPC groups and in the ischemia and pinacidil treatment groups. 7. There is a coincidence between the increase of Cu,Zn-SOD immunoreactivities and the decrease of apoptosis in the presence of ischemia and reperfusion. These results suggest that the protective effects of ishemic preconditioing may related to the SOD activation, and the ischemic preconditioning decreases the apoptosis partially via $K_{APT}$ channel activation in timely reperfused rat skeletal muscle. It is also suggested that inhibition of apoptosis by IPC may related with the SOD activation.

• 대한지역사회영양학회지
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• 제12권4호
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• pp.396-404
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• 2007

Increased oxidative stress contributes to the progression of atherosclerosis. We measured serum antioxidant mineral concentrations, capacities of serum antioxidant enzymes and fasting lipid profile in 97 male patients with coronary artery disease (CAD) and 21 male controls. Nutrient intake was assessed by the semi-quantitative food frequency method. CAD patients were divided into single-vessel disease (SVD, n=66) and multi-vessel disease (MVD, n = 31) groups on the coronary angiography. The ratio of serum LDL- to HDL-cholesterol elevated with an increasing number of diseased vessels compared to the control (control < SVD < MVD, p < 0.05). Patients with SVD and MVD had higher levels of serum lipoprotein (a) than the control (p < 0.05). The mean intake of carbohydrate, protein and cholesterol was higher in MVD patients and the intakes of vitamins C and E were lower in MVD and SVD patients than in the control (p < 0.05). Serum copper (Cu) and zinc (Zn) levels were higher in MVD and SVD patients than in the control (Cu: control $75.8{\pm}5.07$, SVD $99.2{\pm}2.90$, MVD $100.1{\pm}2.32{\mu}g/dL$, p<0.01; Zn: $76.8{\pm}5.36$, $119.0{\pm}5.95$, $129.1{\pm}2.70{\mu}g/dL$, p < 0.01). And the ratio of Zn to Cu was higher in SVD and MVD patients than in the control (control $0.78{\pm}0.06$, SVD $0.88{\pm}0.05$, MVD $0.99{\pm}0.04$, P < 0.05). The activity of glutathione peroxidase (GSH-Px) was lower in MVD than in SVD and the control (control $35.13{\pm}1.34$, SVD $35.30{\pm}1.01$, MVD $31.00{\pm}1.04 U/mg$ protein, p < 0.05). The ratio of the activities of superoxide dismutase (SOD) to GSH-Px was higher in MVD than in control and SVD (p < 0.05). In groups with CAD, serum Cu and Zn concentrations and their ratio were changed compared to the control. GSH-Px activity was decreased and the ratio of SOD to GSH-Px was increased in the patients with MVD. The balances between the activities of SOD and GSH-Px should also be considered a risk factor in CAD patients.

• 생명과학회지
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• 제13권5호
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• pp.658-667
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• 2003

환경 스트레스에 대해 내성을 가지는 것으로 알려진 명아주과에 속하는 근대(지상부길이 15 cm)를 이용하여, 다양한 염 농도에서의 건물함량 측정을 통한 생장반응과 항산화 효소(SOD, APX, GR)의 효과를 밝히기 위하여 다양한 농도(0, 50, 200, 1000 mM NaCl)의 염을 처리한 후 24시간 동안의 효소의 활성변화를 측정하였다. 근대는 처리 2시간째 200 mM NaCl처리구 에서 SOD, APX, GR의 최대활성을 보였으며, 50 mM NaCl처리구에서 가장 낮은 활성을 나타내었다. PAGE에 의한 isoforms의 확인결과, 근대는 3개의 SOD isoforms(Fe-SOD, CuZn-SOD, Mn-SOD)를 함유하고 있었으며, major isoform은 CuZn-SOD로 밝혀졌다. APX의 경우, 9개의 bands 중 특별히 저분자 isoforms (No. 7,8)의 강한 발현양상을 보였다. SOD의 경우 50 mM NaCl처리에서 Mn-SOD isoform의 불활성을 보여 활성의 증감에 있어 Mn-SOD가 직접적인 연관성을 가질 것으로 생각된다. 근대의 항산화 효소는 염 처리후 단시간내 효소 활성의 증가양상(특별히 처리후 2시간째 200 mM NaCl처리구)을 보여, 고농도 염 환경하에서 항산화시스템의 빠른 작동을 통해 염스트레스에 의해 생성된 활성산소를 제거함으로써 염에 의한 산화적 스트레스에 대해 효과적으로 대처해 나가는 것으로 생각된다. 검색어-근대, 염, 활성산소, SOD, APX, GR.

• Molecules and Cells
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• 제20권3호
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• pp.401-408
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• 2005

Reactive oxygen species (ROS) contribute to the development of various human diseases. Cu,Zn-superoxide dismutase (SOD) is one of the major means by which cells counteract the deleterious effects of ROS. SOD activity is dependent upon bound copper ions supplied by its partner metallochaperone protein, copper chaperone for SOD (CCS). In the present study, we investigated the protective effects of PEP-1-CCS against neuronal cell death and ischemic insults. When PEP-1-CCS was added to the culture medium of neuronal cells, it rapidly entered the cells and protected them against paraquat-induced cell death. Moreover, transduced PEP-1-CCS markedly increased endogenous SOD activity in the cells. Immunohistochemical analysis revealed that it prevented neuronal cell death in the hippocampus in response to transient forebrain ischemia. These results suggest that CCS is essential to activate SOD, and that transduction of PEP-1-CCS provides a potential strategy for therapeutic delivery in various human diseases including stroke related to SOD or ROS.

• Journal of Korean Biological Nursing Science
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• 제3권1호
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• pp.41-52
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• 2001

This study was undertaken to investigate the effects of n-6(corn oil) & n-3(fish oil) fatty acids on infarction size and the cerebral activities of antioxidant enzyme in rat focal brain ischemia model. Weaning Sprague-Dawley rats were fed with either corn oil supplemented diet(COD, 14% corn oil) or fish oil supplemented diet(FOD, 14% menhaden oil) for 6 weeks. The right middle cerebral artery was occluded for 2 hours with a silicon rubber coated nylon surgical thread. After 24 hours of recirculation, the rats were sacrificed and brain sections were photographed using CCD camera after staining with 2, 3, 5-triphenyltetrazolium chloride for 60 minutes in room temperature. The infarcted area was measured and the volume of infarction was calculated. Catalase(CAT), superoxide dismutase(SOD) activities, and fatty acid composition in the brain were also measured. The total and corrected infarction volumes were not significantly different between FOD and COD group. The docosagexaenoic acid(DHA) and DHA content/arachidonic acid(AA) ratio of the cerebral cortex, an index of defense against lipid oxidation, were significantly increased in FOD group compared to those of COD group(p<0.05). In the left cortex(non-infarction side) as well as the right cortex(infarction side) of FOD group, CAT and Cu/Zn SOD activities were higher than those of the COD group(p<0.05). However, CAT and Cu/Zn SOD activities were not significantly different between the left cortex(non-infarction side) and the right cortex(infarction side) of both FOD and COD group. GPx activities were also not significantly different between two groups. Our results demonstrate that the brain infarction size in FOD and COD were not significantly different. However, cerebral lipid composition and antioxidant enzyme activities in FOD and COD group were different. Fish oil, a source of n-3 polyunsaturated fatty acid(PUFA) and corn oil, that of n-6(PUFA) may have a protective effect against oxidative stress induced via different mechanisms.

• Korean Journal of Acupuncture
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• 제36권1호
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• pp.63-73
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• 2019

Objectives : This study was designed to investigate the effects of Alismatis Rhizoma (AR) pharmacopuncture at BL23 on acute renal failure induced by cisplatin. Methods : The Sprague-Dawley rats were divided into Normal group (no injection of cisplatin and no treatment), Control group (cisplatin injection without treatment), Acu group (needling at BL23 after cisplatin injection), AR-PA1 group (treated with $0.3571mg/kg/20{\mu}l$ of AR pharmacopuncture at BL23 after cisplatin injection), and AR-PA2 group (treated with $1.7855mg/kg/20{\mu}l$ of AR pharmacopuncture at BL23 after cisplatin injection). Each treatment was given once daily for 8 days. Changes in body weight, kidney weight, tumor necrosis factor-alpha ($TNF-{\alpha}$), interleukin-6 (IL-6), Cu-Zn superoxide dismutase (Cu-Zn SOD), glutathione peroxidase (GPx), serum blood urea nitrogen (serum BUN), and serum creatinine were observed. Results : Body weight was significantly increased in AR-PA1 on $4^{th}$ and $6^{th}$ days and AR-PA2 on $2^{nd}$ day. $TNF-{\alpha}$ was significantly decreased in Acu, AR-PA1 and AR-PA2 groups. Cu-Zn SOD was significantly increased in AR-PA2 group. GPx was significantly increased in AR-PA1 and AR-PA2 groups. But kidney weight, IL-6, serum BUN and serum creatinine were not significantly changed in any groups compared to control group. Conclusions : In acute renal failure induced by cisplatin, AR pharmacopuncture has a mitigating effect on the inflammatory reaction related to the increase of $TNF-{\alpha}$ in the kidney tissue and a protective effect on the oxidative stress of the kidney tissue. However it is unlikely to restore the glomerular function or inhibit the renal swelling.

• 한국약용작물학회지
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• 제27권6호
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• pp.419-426
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• 2019

Background: This study aimed to identify antioxidant compounds from the seeds of Dolichos lablab L. by bioassay-guided isolation and recrystallization. Methods and Results: The water layer of D. lablab L. seed extract inhibits intracellular reactive oxygen species (ROS) expressing the 2',7'-dichlorofluorescein diacetate (DCF-DA), Cu/Zn superoxide dismutase (SOD) and catalase genes, as determined by quantitative real-time PCR (qRT-PCR). Two compounds were purified from the water layer of the seeds of D. lablab L. using column chromatography and prep-high performance liquid chromatography (HPLC). Using nuclear magnetic resonance (NMR) and electrospray Ionization mass spectrometry (ESI-MS), their chemical structures were identified as 5-[(2-acetyl-2,3-dihydro-1H-indazol-1-yl)carbonyl]-4,5-dihydro-3H-furan-2-one (C₁₄H₁₄N₂O₄) and stachyose. Conclusions: Two active antioxidant compounds were purified from the seed extract of D. lablab L. seed extract and the structures of these compounds were identified as C₁₄H₁₄O₄N₂ and stachyose.

• Preventive Nutrition and Food Science
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• 제19권2호
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• pp.82-88
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• 2014

Yam (Dioscorea batatas Decne.) has long been used as a health food and oriental folk medicine because of its nutritional fortification, tonic, anti-diarrheal, anti-inflammatory, antitussive, and expectorant effects. Reactive oxygen species (ROS), which are known to be implicated in a range of diseases, may be important progenitors of carcinogenesis. The aim of this study was to investigate the modulatory effect of yam on antioxidant status and inflammatory conditions during azoxymethane (AOM)-induced colon carcinogenesis in male F344 rats. We measured the formation of aberrant crypt foci (ACF), hemolysate antioxidant enzyme activities, colonic mucosal antioxidant enzyme gene expression, and colonic mucosal inflammatory mediator gene expression. The feeding of yam prior to carcinogenesis significantly inhibited AOM-induced colonic ACF formation. In yam-administered rats, erythrocyte levels of glutathione, glutathione peroxidase (GPx), and catalase were increased and colonic mucosal gene expression of Cu/Zn-superoxide dismutase (SOD), Mn-SOD, and GPx were up-regulated compared to the AOM group. Colonic mucosal gene expression of inflammatory mediators (i.e., nuclear factor kappaB, inducible nitric oxide synthase, cyclooxygenase-2, tumor necrosis factor alpha, and interleukin-1beta) was suppressed by the yam-supplemented diet. These results suggest that yam could be very useful for the prevention of colon cancer, as they enhance the antioxidant defense system and modulate inflammatory mediators.

• Molecules and Cells
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• 제37권2호
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• pp.100-108
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• 2014

Salmonella enterica serovar Typhimurium (S. Typhimurium) is an intracellular pathogen that has evolved to survive in the phagosome of macrophages. The periplasmic copper-binding protein CueP was initially known to confer copper resistance to S. Typhimurium. Crystal structure and biochemical studies on CueP revealed a putative copper binding site surrounded by the conserved cysteine and histidine residues. A recent study reported that CueP supplies copper ions to periplasmic Cu,Zn-superoxide dismutase (SodCII) at a low copper concentration and thus enables the sustained SodCII activity in the periplasm. In this study, we investigated the role of CueP in copper resistance at a high copper concentration. We observed that the survival of a cueP-deleted strain of Salmonella in macrophage phagosome was significantly reduced. Subsequent biochemical experiments revealed that CueP specifically mediates the reduction of copper ion using electrons released during the formation of the disulfide bond. We observed that the copper ion-mediated Fenton reaction in the presence of hydrogen peroxide was blocked by CueP. This study provides insight into how CueP confers copper resistance to S. Typhimurium in copper-rich environments such as the phagosome of macrophages.

• 한국자원식물학회지
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• 제10권2호
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• pp.188-193
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• 1997

고려인삼 1년근에서 5년근을 전기영동하였고 SOD 활성에 대한 염색을 수행하였다. 그 결과 총 13개의 구분되어지는 무색의 밴드를 나타내었다. 그리고 밴드의 패턴은 1년근에서 5년근이 모두 동일 하였다. 이들 효소 중의 9개는 시아나이드 처리나 과산화수소 처리에 의해 밴드가 제거되었고 클로로포름과 에탄올처리에 대해서는 안정했다. 이들 효소들은 CuZnSOD 효소들이다. 반면에 4개의 밴드는 시아나이드처리나 과산화수소 처리에 의해 안정하고 클로로포름과 에탄올 처리에 의해서는 제거되었다. 그러므로 이들은 MnSOD 효소들이다. 인삼 1년근에서 5년근까지 SOD 활성의 비교를 광화학적 에세이 방법에 의해 측정하였다. 이들 SOD 활성 측정의 결과에서 년수에 따른 SOD 활성의 유의성있는 차이는 보여지지 않았다. 모든 인삼 추출물의 총 SOD 활성은 대략 700-800units/g of fresh weight 이었다. 그러나 SOD 효소에 의해 SOD 활성은 대략 200-250 units/g of fresh weight 이었다. 그러므로 SOD 활성 유사 물질에 의해 활성이 SOD 효소에 의한 활성보다 높다. SOD 활성 유사 물질과 SOD 효소에 의한 활성의 비는 대략 2:1에서 3:1이다.