• 대한약침학회지
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• 제20권2호
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• pp.127-131
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• 2017

Objectives: Polymorphonuclear neutrophils (PMNs) constitute the first line of defense against invading microbial pathogens. Early events in inflammation involve the recruitment of neutrophils to the site of injury or damage where changes in intracellular calcium can cause the activation of pro-inflammatory mediators from neutrophils including superoxide generation, degranulation and release of myeloperoxidase (MPO), productions of interleukin (IL)-8 and tumor necrosis factor ${\alpha}$ ($TNF-{\alpha}$), and adhesion to the vascular endothelium. To address the anti-inflammatory role of flavonoids, in the present study, we investigated the effects of the flavonoids quercetin and vitexin on the stimulus-induced nitric oxide (NO), $TNF-{\alpha}$, and MPO productions in human neutrophils. Methods: Human peripheral blood neutrophils were isolated, and their viabilities were determined by using the Trypan Blue exclusion test. The polymorphonuclear leukocyte (PMNL) preparations contained more than 98% neutrophils as determined by morphological examination with Giemsa staining. The viabilities of cultured neutrophils with various concentrations of quercetin and vitexin ($1-100{\mu}M$) were studied using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assays. Neutrophils were cultured in complete Roswell Park Memorial Institute (RPMI) medium, pre-incubated with or without quercetin and vitexin ($25{\mu}M$) for 45 min, and stimulated with phorbol 12-myristate 13-acetate (PMA) ($10^{-7}M$). NO production was carried out through nitrite determination by using the Griess method. Also, the $TNF-{\alpha}$ and the MPO productions were measured using enzyme-linked immunosorbent assay (ELISA) kits and MPO assay kits. Results: Neutrophil viability was not affected up to a concentration of $100{\mu}M$ of quercetin or vitexin. Both quercetin and vitexin significantly inhibited $TNF-{\alpha}$, NO, and MPO productions in human neutrophils (P < 0.001). Conclusion:The present study showed that both quercetin and vitexin had significant anti-inflammatory effects. Thus, treatment with either quercetin or vitexin may be considered as a therapeutic strategy for treating patients with neutrophil-mediated inflammatory diseases.

• 동의생리병리학회지
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• 제21권5호
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• pp.1163-1169
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• 2007

Effects of Sotosaja hwan on Blood Glucose, Hyperlipidemia, Polyol Pathway and Antioxidative Mechanism in ob/ob Mouse Diabetes is a disease in which the body does not produce or properly use insulin. Etiological studies of diabetes and its complications showed that oxidative stress might play a major role. Therefore, many efforts have been tried to regulate free oxygen radicals for treating diabetes and its complications. Sotosaja-hwan has been known to be effective for the antiaging and composed of four crude herbs. In male ob/ob mouse in severe obesity, hyperinsulinemia and hyperlipidemia, which are features of NIDDM, the hyperglycemic activites and mechanisms of Sotosaja-hwan were examined. 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 with a diet of chow supplemented with 30 and 90 mg Sotosaja-hwan per 1 kg of body weight for 14 days. The effects of Sotosaja-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. Sotosaja-hwan lowered the levels of serum glucose and insulin in a dose dependent manner. Total cholesterol, triglyceride and free fatty acid levels were decreased, while the HDL-cholesterol level was increased, in Sotosaja-hwan treated groups. Renal aldose reductase and sorbitol dehydrogenase activities were increased in the ob/ob mice, whereas those were inhibited in the Sotosaja-hwan-administered groups. Sotosaja-hwan inhibited the generation of ${\cdot}O_2$ in the kidney. Finally, MDA+HAE levels was increased and GSH/GSSG ratio was decreased in the ob/ob mice, whereas those were improved in the Sotosaja-hwan-administered groups. Sotosaja-hwan showed the antidiabetic and anti hyperlipidemic activities by regulating the activities of polyol pathway enzymes, scavenging reactive oxygen species and reducing the MDA+HAE levels in the ob/ob mice.

• Nutrition Research and Practice
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• 제8권2호
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• pp.138-145
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• 2014

BACKGROUND/OBJECTIVES: This study was performed to investigate the in vitro antioxidant and cytoprotective effects of fermented sesame sauce (FSeS) against hydrogen peroxide ($H_2O_2$)-induced oxidative damage in renal proximal tubule LLC-PK1 cells. MATERIALS/METHODS: 1,1-diphenyl-2-picrylhydrazyl (DPPH), hydroxyl radical ($^{\bullet}OH$), and $H_2O_2$ scavenging assay was used to evaluate the in vitro antioxidant activity of FSeS. To investigate the cytoprotective effect of FSeS against $H_2O_2$-induced oxidative damage in LLC-PK1 cells, the cellular levels of reactive oxygen species (ROS), lipid peroxidation, and endogenous antioxidant enzymes including catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GSH-px) were measured. RESULTS: The ability of FSeS to scavenge DPPH, $^{\bullet}OH$ and $H_2O_2$ was greater than that of FSS and AHSS. FSeS also significantly inhibited $H_2O_2$-induced ($500{\mu}M$) oxidative damage in the LLC-PK1 cells compared to FSS and AHSS (P < 0.05). Following treatment with $100{\mu}g/mL$ of FSeS and FSS to prevent $H_2O_2$-induced oxidation, cell viability increased from 56.7% (control) to 83.7% and 75.6%, respectively. However, AHSS was not able to reduce $H_2O_2$-induced cell damage (viability of the AHSS-treated cells was 54.6%). FSeS more effectively suppressed $H_2O_2$-induced ROS generation and lipid peroxidation compared to FSS and AHSS (P < 0.05). Compared to the other sauces, FSeS also significantly increased cellular CAT, SOD, and GSH-px activities and mRNA expression (P < 0.05). CONCULUSIONS: These results from the present study suggest that FSeS is an effective radical scavenger and protects against $H_2O_2$-induced oxidative damage in LLC-PK1 cells by reducing ROS levels, inhibiting lipid peroxidation, and stimulating antioxidant enzyme activity.

• 생명과학회지
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• 제22권1호
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• pp.126-131
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• 2012

현미의 기능성을 증명하기 위하여 열수, 에탄올 추출하여 여러 가지 생리활성에 대하여 조사하였다. 우선, 아 질산염 소거능 분석에서는 현미 열수와 에탄올 추출물 1 mg/ml 농도, pH 1.2에서 75.4와 94.9%을 나타내었고, xanthine oxidase 저해능은 현미 에탄올 추출물 10 mg/ml에서 72.9%로 같은 농도의 열수 추출물보다 높게 측정되었다. 현미의 소화율($44.46{\pm}6.09$%)은 백미의 소화율($62.71{\pm}4.18$%)보다 낮으며 ${\alpha}$-glucosidase 저해능은 현미 에탄올 추출물 10 mg/ml에서 93.1%로 같은 농도의 열수 추출물보다 높게 측정되었다. 현미 열수와 에탄올 추출물의 SOD 유사활성은 10 mg/ml에서 56.4와 44.9%로 유의적인 큰 차이는 없었다. 현미의 숙취해소 효능은 ADH와 ALDH 활성증진에 현미 열수와 에탄올 추출물이 미치는 영향을 조사함으로써 증명하고자 하였는데 그 결과, 현미 에탄올 추출물에서 알콜 분해능 증가율이 나타났다.

• The Plant Pathology Journal
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• 제29권4호
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• pp.386-396
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• 2013

Reactive oxygen species (ROS) generation in tomato plants by Ralstonia solanacearum infection and the role of hydrogen peroxide ($H_2O_2$) and nitric oxide in tomato bacterial wilt control were demonstrated. During disease development of tomato bacterial wilt, accumulation of superoxide anion ($O_2{^-}$) and $H_2O_2$ was observed and lipid peroxidation also occurred in the tomato leaf tissues. High doses of $H_2O_2$ and sodium nitroprusside (SNP) nitric oxide donor showed phytotoxicity to detached tomato leaves 1 day after petiole feeding showing reduced fresh weight. Both $H_2O_2$ and SNP have in vitro antibacterial activities against R. solanacearum in a dose-dependent manner, as well as plant protection in detached tomato leaves against bacterial wilt by $10^6$ and $10^7$ cfu/ml of R. solanacearum. $H_2O_2$- and SNP-mediated protection was also evaluated in pots using soil-drench treatment with the bacterial inoculation, and relative 'area under the disease progressive curve (AUDPC)' was calculated to compare disease protection by $H_2O_2$ and/or SNP with untreated control. Neither $H_2O_2$ nor SNP protect the tomato seedlings from the bacterial wilt, but $H_2O_2$ + SNP mixture significantly decreased disease severity with reduced relative AUDPC. These results suggest that $H_2O_2$ and SNP could be used together to control bacterial wilt in tomato plants as bactericidal agents.

• Preventive Nutrition and Food Science
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• 제5권4호
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• pp.213-218
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• 2000

The purpose of the study was to investigate the effects of dietary green tea catechin and vitamin E on the phospholipse {TEX}$A_{2}${/TEX} activity and th antioxidative defense system in streptozotocin (STZ)-induced diabetic rats. Sprague-Dawley male rats weighing 100$\pm$10 gm were randomly assigned to one normal and five STZ-induced diabetic groups. The diabetic groups were assigned either a catechin-free diet (DM group), 0.5% catechin diet (DM-0.5C group), 1% catechin diet (DM-1C group), vitamin E-free diet (DM-0E group), and 400 mg vitamin E per kg diet (DM-400E group) according to the levels of dietary catechin or vitamin E supplementation. The vitamin E levels of the normal, DM, DM-0.5C, and DM-1C groups were 40 mg per kg diet. Diabetes was experimentally induced by an intravenous injection of streptozotocin after 4 weeks of feeding the five experimental diets. The animals were sacrificed on the 6th day of he diabetic state. The body weight gains were lower in all five diabetic groups after the STZ injection. The platelet phospholipase {TEX}$A_{2}${/TEX}({TEX}$PLA_{2}${/TEX}) activity in the diabetic groups was higher than that in the normal group. However, the enzyme activity in the DM-0.5C, DM-1C, and DM-400E groups was lower than that in the DM and DM-0E groups. The cytochrome {TEX}$P_{450}${/TEX} and cytochrome {TEX}$b_{5}${/TEX} content and NADPH-cytochrome {TEX}$P_{450}${/TEX} reductase activity were about 50~110% higher in the DM and DM-0E groups than in the normal group, yet significantly reduced by either catechin or vitamin E supplementation. The superoxide dismutase (SOD) content in the liver did not differ significantly in any of the groups. However, the glutathione peroxidase (GSHpx) activity was generally lower in the diabetic groups, compared with the normal group, whereas that of the DM-0.5C, DM-1C, and DM-400E groups was significantly higher compared with that of the DM and DM-0E groups. The levels of thiobarbituric acid reactive substances (TBARS) in the liver tissue were 148% and 201% higher in the DM and DM-0E groups, respectively, compared with the normal group, however, these levels were reduced by either catechin or vitamin E supplementation (DM-0.5, DM-1C and DM-400E). Accordingly, the present results indicate that STZ-induced diabetic rats exhibited an imbalance between free radical generation and scavenger systems in the liver which led to the acceleration of lipid peroxidation. However, these abnormalities were reduced and the antioxidative defense system was restored by either dietary catechin or vitamin E supplementation. In conclusion, the effects of dietary catechin or vitamin E in streptozotocin-induced diabetic rats would appear to inhibit lipid peroxidation as an anti-oxidant by regulating the activity of {TEX}$PLA_{2}${/TEX}.

• BMB Reports
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• 제31권2호
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• pp.161-169
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• 1998

TThe reactive oxygen species oxidatively modify the biological macromolecules, including proteins, lipids, and nucleic acids. Iron- and heme-mediated Fenton-like reactions produce different pro-oxidants. However, these reactive products have not been clearly characterized. We examined the nature of the oxidizing species from the different iron sources by measuring oxidative protein modification and spectroscopic study. Hemoglobin (Hb) and methemoglobin (metHb) were oxidatively modified in $O{\array-\\\dot{2}}$ and $H_{2}O_{2}$ generating systems. Globin and bovine serum albumin (BSA) were also modified by iron, iron-EDTA, hematin, and Hb in an $O{\array-\\\dot{2}}$ generating system. In a $H_{2}O_{2}$ generating system, the iron- and iron-EDTA-mediated protein modifications were markedly reduced while the Hb-and hematin-mediated modifications were slightly increased. In the $O{\array-\\\dot{2}}$ generating system, the iron- and iron-EDTA-mediated protein modifications were strongly inhibited by superoxide dismutase (SOD) or catalase, but heme- and Hb-mediated protein modifications were inhibited only by catalase and slightly increased by SOD. Mannitol, 5,5-dimethyl-l-pyrroline-N-oxide (DMPO), deoxyribose, and thiourea inhibited the iron-EDTA-mediated protein modification. Mannitol and DMPO, however, did not exhibit significant inhibition in the hematin-mediated modification. Desferrioxamine (DFO) inhibited protein modification mediated by iron, but cyanide and azide did not, while the hematin-mediated protein modification was inhibited by cyanide and azide, but not significantly by DFO. The protein-modified products by iron and heme were different. ESR and UV-visible spectroscopy detected the DMPO spin adduct of the hydroxyl radical and ferryl ion generated from iron-EDTA and metHb, respectively. These results led us to conclude that the main oxidizing species are hydroxyl radical in the iron-EDTA type and the ferry I ion in the hematin type, the latter being more effective for protein modification.

• 한국식품영양과학회지
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• 제34권9호
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• pp.1330-1335
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• 2005

붉은 갈색계통 염색제 성분 중의 하나인 p-phenylenediamine(PPD)는 일반적으로 여성들이 사용하는 염색제의 주요 성분으로 시야 흐림 및 구토와 같은 전신적 아나필락시스, 피부염 또는 방광암 등이 유발된다고 한다. 그러나 PPD의 피부 독성과 유해산소와 관련된 연구는 아직까지 미흡한 실정이다. 본 실험에서는 체중 $230\pm20g$의 Sprague-Dawley 종의 흰쥐의 피부에 PPD을 도포하였을 때 조직의 손상정도를 상호비교하고 조직의 손상 원인을 구명하고자 실험동물에 PPD($2.5\%$ PPD in $2\%\;NH_{4}OH$)을 표면적 $25 mg/16.5\;cm^2$씩 2일 간격으로 3회 및 5회 도포하였다. PPD 3회보다 5회 도포군에서 피부조직 의 표피 및 케라틴 층의 비후, glucose 5-phosphatase 활성의 감소 및 acid phosphatase 활성의 증가 정도가 높게 나타났다. 또한 유해산소의 생성계 및 해독계와 관련하여 피부조직의 손상정도를 확인한 결과, 유해산소 생성계인 xanthine oxidase의 활성은 PPD 3회 보다5회 도포군에서 유의하게 증가하였다. 그러나 유해산소 해독계인 superoxide dismutase, catalase, glutathione S-transferase 활성 및 reduced glutathione 함량은 대조군보다 PPD 도포군에서 감소하였다. 한편, xanthine oxidase의 활성이 3회 PPD 도포군에서는 별다른 변동이 없음에도 불구하고 oxygen free radical system의 감소와 피부조직의 손상이 나타난 것은 흰쥐의 피부조직 에서 PPD를 대사하는 동안 생성된 대사산물에 의한 것으로 생각된다. 이상의 실험 결과를 종합해 볼 때 PPD 도포에 의한 피부조직의 손상은 PPD 도포 횟수에 비례하여 유해산소 생성율이 증가하여 나타난 결과로 생각된다.

• 한국식품과학회지
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• 제36권6호
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• pp.968-973
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• 2004

나물이나 전분원료로 사용되어 온 얼레지(Erythronium japonicum)의 항암작용을 검색하였다. 얼레지 추출물을 Sarcoma 180으로 복수암을 유발시킨 ICR 마우스에 대하여 경구투여 한 결과 143.7%에 이르는 생명연장효과를 얻었으며, 백혈병계 암세포인 L1210 세포에 대해서는 최고 98.6%의 세포수 감소효과를 얻었다. 아울러 얼레지 추출물의 독성유무를 검색하고자 normal lymphocyte를 분리하여 이 정상세포에 얼레지 추출물을 첨가했을 때 세포수 감소는 거의 일어나지 않았으며, 고농도의 3일간의 긴 배양기간에 의해서도 L1210 세포의 경우 83.2%에 비해 5.8% 정도로 매우 적었다. 이와 같은 결과로부터 얼레지 추출물은 무독성 또는 저독성 항암제로서의 개발 가치가 있을 것으로 사료되었다. 한편 얼레지 추출물의 암세포에 대한 작용 기작으로 일환으로 활성산소인 ${O_2}^-$의 생성량을 측정한 결과 control 값의 최대 약 3.6배까지 크게 증가하였으며, 동시에 ${O_2}^-$ 전환효소인 SOD와 SOD의 종산물인 $H_2O_2$, 분해효소인 GPx도 각각 5배와 3배까지 크게 증가하였다. 따라서 얼레지 추출물은 암세포에서 ${O_2}^-$를 비롯한 활성산소를 유발시키고 이 증가된 활성산소가 암세포의 apoptosis를 야기하는 것으로 간주되었다.

• 동의생리병리학회지
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• 제19권6호
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• pp.1557-1562
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• 2005

A mechanism of hair cell damage caused by noise and ototoxic agents is mediated through generation of free radicals and reactive oxygen species (ROS). It is known that most of animals have defense systems to protect against ROS, and the cochlea of inner ear in animals also has ROS defense systems including several antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX), glutathione reductase (GR), and glutathione (GSH), which efficiently detoxifying ROS generated under normal condition. Steamed roots of Rehmannia glutinosa have been traditionally used in Oriental medicine for the treatment of auditory disease such as tinnitus, vertigo, and hearing loss as well as inflammatory diseases, hectic fever, night sweat, and headache. In the present study, we showed that the ethanol extract from steamed roots of R. giutinosa (ESRG) increased the antioxidant enzymes such as SOD, CAT, GPX, and GR activities and GSH level in HEI-OC1 auditory cells. This extract itself did not show any significant cytotoxicity up to $50{\mu}g/ml$. Our results further support the view that ESRG is promising sources of potential antioxidants. Future studies will be aimed at investigating the effects of ESRG on the regulation of cellular mechanisms and isolating and identifying the substances responsible for the regulation of antioxidant enzyme system from the plant extracts.