• Tuberculosis and Respiratory Diseases
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• 제65권6호
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• pp.464-470
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• 2008

연구배경: 산소독에 의한 급성 폐손상에 대한 기전은 아직 확실히 알려져 있지 않다. 급성호흡곤란증후군에서 보이는 호중구의 산소기 생성에 따른 조직의 손상이 산소독에 의한 손상에서도 관여하는지를 확인하고자 하였다. 방 법: Plastic cage 내의 기압을 1기압으로 고정하고 흰쥐에게 순수한 산소를 48시간 호흡시킨 후 호중구가 폐장 내로 침윤함으로써 나타나는 급성 폐손상을 생화학적인 지표 및 형태학적인 관찰 등을 통하여 검사하였다. 결 과: 흰쥐에게 순수한 산소를 48시간 호흡하게 한 경우 폐부종, 호중구의 침윤, 폐장 내 MDA 및 $cPLA_2$ 활동도의 증가가 관찰되었고, 형태학적으로도 탐식구 특히 호중구의 침윤에 따른 폐장의 손상이 관찰되었다. 결 론: 산소독에 의한 급성폐손상의 기전은 고농도의 산소에 의한 산소기 생성이 그 원인으로 생각 되지만 산소기의 작용기전은 부분적으로 $cPLA_2$활성도 증가에 의한 호중구의 조직 내 침윤에 따른 이차적 산소기 형성이 그 원인으로 생각되며 시간이 경과할수록 호중구에 폐장 내 침윤에 따른 손상이 더 심해질 것으로 생각된다.

• Tuberculosis and Respiratory Diseases
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• 제41권3호
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• pp.222-230
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• 1994

연구배경 : Paraquat는 광범위 제초제로서 널리 사용하고 있으나, 중독시 치명적인 중독 증상을 일으킨다. 특히 폐에서는 진행성 불가역성 폐섬유종을 일으키는데, 이의 기전으로 산소유리기와 관계가 있으나 아직은 생화학적 기전이 명확하지 않다. paraquat에 의한 산소유리기가 생기면 glutathione의 변화와 G6PDH, SOD, catalase 및 glutathione peroxidase등의 효소 활성의 변화가 생길 것으로 생각되며, 특히 catalase가 많이 관여할 것으로 생각되어진다. 방법 : Catalase 억제제인 aminotriazole을 사용하여 paraquat만 쓰는 것과 paraquat와 aminotriazole을 같이 투여할때 생쥐의 생존율을 알아보고, 실험군을 정상대조군, Group A(aminotriazole투여군), Group B(paraquat 투여군), Group C(paraquat와 amino-triazole 병합투여군) 4군으로 나누어 폐조직에서 glutathione량, G6PDH, SOD, catalase 및 glutathione peroxidase활성도를 측정하여 비교했다. 결과 : Paraquat와 aminotriazole 병합 투여군의 생존율이 paraquat투여군보다 현저히 감소하였고, paraquat 투여로 인하여 폐 glutathione량은 정상대조군에 비해 20%정도 감소 되었으나, aminotriazole의 투여로 인한 폐 glutathione량의 변화는 없었다. Paraquat투여로 폐 SOD, catalase 및 glutathione peroxidase활성이 모두 유의한 감소를 나타냈는데, 특히 catalase가 가장 큰 효소활성 감소를 나타냈으며, paraquat와 aminotriazole병합투여군에서는 catalase와 glutathione peroxidase활성이 paraquat단독투여군에 비하여 유의한 효소활성감소를 나타냈고, SOD는 효소활성의 변화가 감지되지 않았다. 결론 : Paraquat투여시 catalase활성이 유의하게 감소되는 점으로 보아 paraquat독성이 catalase활성과 밀접하게 연관되는 것으로 사료되며, 또한 paraquat의 독성이 aminotriazole의 병합 투여로 더욱 증가되어 나타나는데, 이러한 결과는 aminotriazole투여로 catalase활성의 감소가 크게 나타나나 glutathione량의 변화는 없는 점으로 보아 aminotriazole투여에 의한 paraquat독성의 증가는 총 폐 glutathione량의 변화에 의한 영향보다는 catalase활성감소에 의한 결과로 생각된다. Paraquat와 aminotriazole를 병합 투여하여 catalase활성이 억제되면 증가된 과산화수소로 hydroxyl radicals이 생성되고, 이에 의한 폐 세포손상이 유발되어 나타나는 것으로 사료된다.

• 생명과학회지
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• 제24권10호
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• pp.1078-1084
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• 2014

본 연구는 꽈리허리노린재(Acanthocoris sordidus) 추출물이 산화적 스트레스에 의해 유발되는 세포와 DNA의 손상 억제력을 조사하였다. 꽈리허리노린재 추출물의 DPPH 유리 라디칼과 수산화 라디칼 제거능은 양성 대조군에 비해 각각 48.9%, 37.8%로 나타났으며, $Fe^{2+}$-chelating 효과는 80.0%로 조사되었다. 꽈리허리노린재 추출물이 활성산소에 의해 유도되는 세포손상에 미치는 억제 효과를 조사하기 위해 지질과산화의 상대적 수준과 p21 단백질의 발현율을 조사해보면 꽈리허리노린재 추출물은 라디칼 처리군에 비해 지질과산화를 거의 완벽하게 억제하고 있으며, p21 단백질의 발현은 대조군의 92.4%로 회복되는 것으로 조사되었다. 또한 꽈리허리노린재 추출물의 DNA 분절화 억제 활성은 대조군에 비해 53.3%로 나타나 산화적 스트레스에 의해 유발되는 DNA 분절화를 효율적으로 억제하고 있으며, H2AX 단백질의 인산화비는 라디칼 처리군의 39.0%에 해당하는 수준으로 조사되어 꽈리허리노린재 추출물은 히스톤 단백질의 인산화를 매우 효율적으로 억제하는 것으로 확인되었다. 이러한 결과로 보아 꽈리허리노린재 추출물은 활성산소에 대한 항산화 효과뿐만 아니라 세포와 DNA의 손상을 억제하는데 효과적인 것으로 사료된다.

• 대한한의학회지
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• 제28권3호통권71호
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• pp.57-69
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• 2007

Objectives : Diabetes is a disease in which the body does not produce or properly use insulin. Etiological studies of diabetes and its complications have shown that oxidative stress might play a major role. Therefore, many methods have been tried to regulate free oxygen radicals for treating diabetes and its complications. Ojung-hwan, composed of five crude herbs, has been considered effective for treating symptoms of aging. In male ob/ob mouse of severe obesity, hyperinsulinemia and hyperlipidemia, which are features of NIDDM, the hyperglycemic activities and mechanisms of Ojung-hwan were examined. Methods : Mice were grouped and treated for 5 weeks as follows. Both the lean (C57/BL6J black mice) and diabetic (ob/ob mice) control groups received standard chow. The experimental groups were fed a diet of chow supplemented with 30 and 90 mg Ojung-hwan per 1 kg of body weight for 14 days. The effects of Ojung-hwan extract on the ob/ob mice were observed by measuring the serum levels of glucose, insulin, lipid components, and the kidney levels of superoxide anion radical (${\cdot}\;O{_2}{^-}$), MDA+HAE, GSH/GSSG ratio, and also the enzyme activities involved in polyol pathway. Results : Ojung-hwan lowered the levels of serum glucose and insulin in a dose-dependent manner. Total cholesterol, triglyceride and free fatty acid levels decreased, while the HDL-cholesterol level increased, in Ojung-hwan treated groups. Renal aldose reductase and sorbitol dehydrogenase activities increased in the ob/ob mice, whereas they were inhibited in the Ojung-hwan treated groups. Ojung-hwan inhibited the generation of ${\cdot}\;O{_2}{^-}$ in the kidney. Finally, MDA+HAE levels increased and GSH/GSSG ratio decreased in the ob/ob mice, whereas they improved in the Ojung-hwan treated groups. Conclusions : Ojung-hwan showed antidiabetic and antihyperlipidemic activities by regulating theactivities of polyol pathway enzymes, scavenging reactive oxygen species and reducing the MDA+HAE levels in the ob/ob mice.

• Archives of Pharmacal Research
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• 제26권4호
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• pp.279-285
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• 2003

In our ongoing study to identity antioxidants from natural sources, the antioxidant activity of Nelumbo nucifera stamens was evaluated for their potential to scavenge stable 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radicals, inhibit total reactive oxygen species (ROS) generation, in kidney homogenates using 2 ,7 -dichlorodihydrofluorescein diacetate (DCHF-DA), and scavenge authentic peroxynitrites ($ONOO^-$). A methanol (MeOH) extract of the stamens of N. nucifera showed strong antioxidant activity in the $ONOO^-$system, and marginal activity in the DPPH and total ROS systems, so were therefore fractionated with several organic solvents, such as dichloromethane ($CH_2 Cl_2$), ethyl acetate (EtOAc) and n-butanol (n-BuOH). The EtOAc soluble fraction, which exhibited strong antioxidant activity in all the model systems tested, was further purified by repeated silica gel and Sephadex LH-20 column chromatographies. Seven known flavonoids [kaempferol (1), kaempferol 3-Ο-$\beta$-D-glucuronopyranosyl methylester (2), kaempferol 3-Ο-$\beta$-D-glucopyranoside (3), kaempferol 3-Ο-$\beta$-D-galactopyranoside (4), myricetin 3 ,5 -dimethylether 3-Ο-$\beta$-D-glucopyranoside (5), kaempferol 3-Ο-$\alpha$-L-rhamnopyranosyl-(1$\rightarrow$6)-$\beta$-D-glucopyranoside (6) and kaempferol 3-Ο-$\beta$-D-glucuronopyranoside (7)], along with $\beta$-sitosterol glucopyranoside (8), were isolated. Compound 1 possessed good activities in all the model systems tested. Compounds 2 and 7 showed scavenging activities in the DPPH and $ONOO^-$ tests, while compounds 3 and 4 were only active in the $ONOO^-$ test. Conversely, compound 8 showed no activities in any of the model systems tested.

• 동의신경정신과학회지
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• 제11권1호
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• pp.1-18
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• 2000

To study the effects of Ramulus et Uncus Uncariae (REUU) on oxygen free radical-mediated damage by hydrogen peroxide $(H_{2}O_{2})$ on cultured spinal sensory neurons, in vitro assays such as MTT assay, NR assay, neurofilament enzymeimmuno assay (EIA), sulforhodamine B (SRB) assay, assay for lactate dehydrogenase (LDH) activity and assay for lipid peroxidation were used in cultured spinal dorsal root ganglion neurons derived from mice, Spinal dorsal root ganglion neurons were cultured in media containing various concentrations of $H_{2}O_{2}$ for 5 hours, after which the neurotoxic effect of $H_{2}O_{2}$ was measured by in vitro assay. The protective effect of the herb extract, Ramulus et Uncus Uncariae (REUU) against H2O2-induced neurotoxicity was also examined. The results are as follows. 1. In NR assay and MTT assay, $H_{2}O_{2}$ significantly decreased the cell viability of cultured mouse spinal dorsal root ganglion neurons according to exposure concentration in these cultures. An additional time course study was done on these cultures. 2. Cultured spinal dorsal root ganglion neurons which were exposed to various concentrations of $H_{2}O_{2}$ showed a quantitative decrease of neuronal cells by EIA and of total protein by sulforhodamine B (SRB) assay, while they showed an increase of both lipid peroxidation and LDH activity. 3. The effect of Ramulus et Uncus Uncariae (REUU) on $H_{2}O_{2}$ induced neurotoxicity showed a quantitative increase in both neurofilament and total protein, but showed a decrease of lipid peroxidation and LDH activity. These results suggest that $H_{2}O_{2}$ has a neurotoxic effect on cultured spinal dorsal root ganglion neurons from mice and that the herb extract, Ramulus et Uncus Uncariae (REUU), was very effective in protecting $H_{2}O_{2}$ induced neurotoxicity by decreasing lipid peroxidation and LDH activity.

• Archives of Pharmacal Research
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• 제19권3호
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• pp.178-182
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• 1996

Using superoxide dismutase (SOD)-deficient mutants of Saccharomyces cerevisiae, the oxidative stresses induced by 0.1 mM of copper ion $(Cu^{++})$ was studied. In aerobic culture condition, yeasts lacking MnSOD (mitochondrial SOD) showed more significant growth retardation than CuZnSOD (cytoplasmic SOD)-deficient yeasts. However, not so big differences in growth pattern of those mutants compared withwild type were observed under anaerobic condition. It was found that, under aerobic condition, the supplementation of 0.1 mM copper ioh:(Cu") into culture medium caused the remarkable increase of CuZnSOD but not so significant change in MnSOD. It was also observed that catalase activities appeared to be relatively high in the presence of copper ion in spite of the remarkable reduction of glutathion peroxidase in CuZnSOD-deficient yeasts, but the slight increments of catalase and glutathion peroxidase were detected in MnSOD-deficient strains. It implies that the lack of cytoplasmic SOD could be compensated mainly by catalase. However, these phenomena resulted in the significantincrease of cellular lipid peroxides content in CuZnSOD-deficient yeasts and the slight increment of lipid peroxides in MNSOD-deficient cells. In anaerobic cultivation supplementing copper ion, the cellular enzyme activities of catalase and glutathion peroxidase in SOD-deficient yeasts were slightly increased without any significant changes of lipid peroxides in cell membrane. It suggests that a little amount of free radicals generated by copper ion under anaerobic condition could be sufficiently overcome by catalase as well as glutathion peroxidase.dase.

• Tuberculosis and Respiratory Diseases
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• 제42권2호
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• pp.184-205
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• 1995

연구배경: Bleomycin(BLM) 투여로 유발되는 폐손상 및 폐섬유화는 특발성 폐섬유증의 실험적 모델로써 BLM의 폐독성 기전은 산소유리기에 의한 직접 독성과 염증세포, 섬유모세포 및 세포외 기질에 의한 간접 독성으로 나누어 설명하고 있다. 이에 저자는 폐손상의 주요 원인이 되는 산소유리기의 생성과 이미 형성된 산소유리기를 보집할 수 있는 산화방지제들을 투여함으로써, BLM에 의한 폐손상 및 폐섬유화의 발생과 그 억제기전을 이해하고자 본 실험을 시행하였다. 방법: 웅성 백서에 폐손상 및 폐섬유화의 대표적인 실험적 방법인 bleomycin (BLM)을 투여함과 동시에 산화방지제로 구리의 킬레이트제인 penicillamine, 철의 킬레이트제인 deferoxamine과 산소유리기의 보집제인 vitamine E를 투여한 후 그 형태학적 변화를 보기 위해 기관지폐포 세척액 검사, 광학현미경 소견과 세포외 기질인 교원섬유 제 I, III, IV형과 비교원성 기질인 fibronectin과 laminin 그리고 근섬유모세포에 대한 NBD phallicidin에 대한 면역형광염색 및 면역조직화학염색소견을 관찰하여 다음과 같은 결과를 얻었다. 결과: 1) 기관지폐포세척액의 검사소견상 총 세포수는 BLM 단독투여군의 경우 제4일에 최고값에 도달한 후 감소하기 시작하여 제28일에 대조군과 비슷해졌지만, penicillamine과 deferoxamine 처치군은 제1일부터 제28일까지 계속 높았으며, vitamin E 처치군은 제3일부터 현저한 차이로 감소하여 제7일부터 대조군과 비슷해졌다. 기관지폐포 세척액의 각 염증세포구성비는 BLM 단독투여군은 초기 즉 제1, 3, 4일에 중성구가 현저히 증가하다가 제7일부터 감소하여 제28일에 대조군과 비슷해졌지만, vitamin E 처치군은 중성구가 제3일부터 BLM 단독투여군의 1/3이하로 현저히 감소하였다. 2) 광학현미경소견은 BLM 단독투여군의 경우 다른 실험군에 비해 섬유화가 가장 심하였고, penicillamine 처치군은 중성구의 침윤이 지속적으로 많았으며, deferoxamine 처치군은 폐포 중격의 림프구 침윤이 뚜렷하였고, vitamin E 처치군은 페포 중격 및 폐포강내 대식세포 증가가 많았다. 이러한 소견은 기관지폐포세척액 소견과 잘 일치하였다. 산화방지제 처치군의 섬유화 소견은 deferoxamine 처치군이 가장 뚜렷하였고, penicillamine 처치군은 적었지만 관찰되었으며, vitamin E 처치군은 거의 없었다. 3) 세포외 기질의 변화로 폐포 간질의 기저막에 양성반응을 나타내는 제 IV형 교원섬유와 laminin 및 Jones' methenamine silver 염색결과는 비슷한 양상이었는데, BLM 단독투여군, penicillamine 처치군과 deferoxamine 처치군은 모두 제28일군에서 음성반응을 나타냈으나, vitamin E 처치군은 제28일까지 대조군과 비슷하게 기저막이 유지되었다. 섬유화 부위에 증가된 세포외 기질은 제 I 형 교원섬유와 fibronectin 이었으며, 제 III 형 교원섬유는 각 국에서 초기에 일시적으로 양성반응을 나타내었다. NBD phallicidin은 BLM 단독투여군에서 특히 제 28일군 육아조직의 방추형세포에 양성반응을 보였다. 결론: Bleomycin 투여에 의한 폐손상 및 폐섬유화 기전은 bleomycin이 구리 또는 철 이온의 존재하에 생성한 산소유리기에 의해 유발될 수도 있으나, 주로 중성구의 활성화로 인한 산소유리기에 의해 유발되고, 폐의 섬유화 제 I 형 교원섬유와 fibronectin의 증가로 초래되었다. 폐손상의 억제는 킬레이트제 보다는 산소유리기의 보집제인 vitamin E 투여시 가장 효과가 있었는데 이것은 초기의 중성구 증가 및 중성구의 활성화 정도에 따라 간질내 폐포의 기저막 유지 및 재생이 가능하고 이것이 폐의 탄력성을 유지시켜 폐손상을 억제할 수 있기 때문으로 사료된다.

• Journal of Periodontal and Implant Science
• /
• 제25권1호
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• pp.167-178
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• 1995

It has been believed that the increased release of free oxygen radicals and their tissue damaging potency might be a contributing factor in the pathogenesis of periodontal disease. Antioxidant enzymes such as superoxide dismutase(SOD) and catalase can protect the tissue damage from the free oxygen radicals($O_2^-,H_2O_2$, and $OH^-$). In order to investigate the SOD- and catalase - activity in the blood plasma and red blood cells(RBCs) of the patients with perodontitis, 19 male periodontitis patients($25{\sim}35$ years old) who had good general health, more than 10 teeth with severely inflamed gingiva, attachment loss more than 6mm and bone loss were selected as periodontitis group, and 13 male volunteers($22{\sim}29$ years old) with good general and periodontal health were selected as normal group. After blood plasma and RBC were separated from peripheral blood of 2ml collected from antecubital vein of each subject, SOD- activity in blood plasma and RBCs was measured by the same method that Paoletti et al. did, and catalase - activity in RBC was measured by the same method that Beers et al, did. The difference of SOD- and catalase - activity between the normal and the periodontitis groups was statistically analyzed by Student t-test with SPSS/PC program.The results were as follows : 1. SOD activity in blood plasma was significantly lower in the periodontitis group($1.986{\pm}0.893$) than in the normal group($3.324{\pm}1.044$)(p<0.05). 2. There was no statistical significance in the difference of SOD- activity in RBCs between the periodontitis group($7.753{\pm}3.206$) and the normal group($8.116{\pm}1.192$)(p$242.8{\pm}45.6$) than in the normal group($280.2{\pm}32.6$)(p

• The Korean Journal of Physiology and Pharmacology
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• 제3권4호
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• pp.393-404
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• 1999

We have reported that hypoxia stimulates EDRF(s) release from endothelial cells and the release may be augmented by previous hypoxia. As a mechanism, it was hypothesized that reoxygenation can stimulate EDRF(s) release from endothelial cells and we tested the hypothesis via bioassay experiment. In the bioassay experiment, rabbit aorta with endothelium was used as EDRF donor vessel and rabbit carotid artery without endothelium as a bioassay test ring. The test ring was contracted by prostaglandin $F_{2a}\;(3{\times}10^{-6}\;M)$ which was added to the solution perfusing through the aorta. Hypoxia was evoked by switching the solution aerated with 95% $O_2/5%\;CO_2$ mixed gas to one aerated with 95% $O_2/5%\;CO_2$ mixed gas. Hypoxia/reoxygenation were interexchanged at intervals of 2 minutes (intermittent hypoxia). In some experiments, endothelial cells were exposed to 10-minute hypoxia (continuous hypoxia) and then exposed to reoxygenation and intermittent hypoxia. In other experiments, the duration of reoxygenation was extended from 2 minutes to 5 minutes. When the donor aorta was exposed to intermittent hypoxia, hypoxia stimulated EDRF(s) release from endothelial cells and the hypoxia-induced EDRF(s) release was augmented by previous hypoxia/reoxygenation. When the donor aorta was exposed to continuous hypoxia, there was no increase of hypoxia-induced EDRF(s) release during hypoxia. But, after the donor aorta was exposed to reoxygenation, hypoxia-induced EDRF(s) release was markedly increased. When the donor aorta was pretreated with nitro-L-arginine $(10^{-5}$ M for 30 minutes), the initial hypoxia-induced EDRF(s) release was almost completely abolished, but the mechanism for EDRF(s) release by the reoxygenation and subsequent hypoxia still remained to be clarified. TEA also blocked incompletely hypoxia-induced and hypoxia/reoxygenation-induced EDRF(s) release. EDRF(s) release by repetitive hypoxia and reoxygenation was completely blocked by the combined treatment with nitro-L-arginine and TEA. Cytochrome P450 blocker, SKF-525A, inhibited the EDRF(s) release reversibly and endothelin antgonists, BQ 123 and BQ 788, had no effect on the release of endothelium-derived vasoactive factors. Superoxide dismutase (SOD) and catalase inhibited the EDRF(s) release from endothelial cells. From these data, it could be concluded that reoxygenation stimulates EDRF(s) release and hypoxia/reoxygenation can release not only NO but also another EDRF from endothelial cells by the production of oxygen free radicals.