• 제목/요약/키워드: superoxide generation

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Inhibitory Effect of Panax notoginseng on Nitric Oxide Synthase, Cyclo-oxygenase-2 and Neutrophil Functions

  • Park, Soon-Gi;Joo, Shin-Tak;Ban, Chang-Gyu;Moon, Jin-Young;Park, Sun-Dong;Park, Won-Hwan
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.20 no.5
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    • pp.1295-1302
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    • 2006
  • Our preliminary aim is to elucidate the pharmacokinetic features of the PNS(Panax notoginseng Buck F.H. Chen. (Arialiaceae) root). First, we assessed the prevention of neurtrophil functions. A Panax notoginseng inhibited neutrophil functions, including degranulation, superoxide generation, and leukotriene B4 production, without any effect on 5-lipoxygenase activity. This Panax notoginseng reduced nitric oxide (NO) and prostaglandin E2 production in mouse peritoneal macrophages stimulated with lipopolysaccharide, whereas no influence on the activity of inducible NO synthase, cyclo-oxygenase-2 or cyclo-oxygenase-1 was observed. Panax notoginseng significantly reduced mouse paw oedema induced by carrageenan. The results indicate that Panax notoginseng exerts anti-inflammatory effects related to the inhibition of neutrophil functions and of NO and prostaglandin E2 production, which could be due to a decreased expression of inducible NO synthase and cyclo-oxygenase-2.

Anticancer Activity of Monoterpenes and the Changes of Enzymes Activities Responsible for the Conversion of Reactive Oxygen Species (Monoterpenes의 항암작용과 활성산소 전환 효소의 활성 변화)

  • 조용선;김수진;박시원
    • YAKHAK HOEJI
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    • v.47 no.1
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    • pp.37-45
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    • 2003
  • The present study was undertaken to investigate the anticancer activity of monoterepenes in the animal and the cancer cell line tests. Both of the noncyclic and cyclic monoterpenes showed significant life prolonging effects on ICR mouse with abdominal cancer induced by Sarcoma 180 cells up to 67.4% and 63.5% in case of linalool and geraniol, respectively. Linalool and geraniol also exhibited very excellent cytotoxicity against L1210 leukemic cells with $IC_{50}$/ value of 0.32 $\mu\textrm{g}$/mι in 5 days culture condition. In the presence of linalool and geraniol, the generation of $O_2$$^{[-10]}$ ion were found to be increased proportionally to the cytotoxicity arisen from these monoterpenes. Furthermore, the antioxidant enzymes activities such as superoxide dismutase (SOD) and glutathione peroxidase (GPx) responsible for the conversion of $O_2$$^{[-10]}$ ion to $H_2O$$_2$ and then to $H_2O$ augmented remarkably by linalool and geraniol. All data put together it can be postulated that monoterpenes may kill abdominal cancer cells of ICR mouse probably by activating anticancer system of the body, whereas the death of L1210 cells may be due to the detrimental attacks of reactive oxygen species (ROS) including $O_2$$^{[-10]}$ in spite of antioxidant enzymes activities to overcome the ROS attacks.

Protective effects of Erythronium japonicum and Corylopsis coreana Uyeki extracts against 1,3-dichloro-2-propanol-induced hepatotoxicity in rats

  • Seunghyun Kim;Hee-Ock Boo;Taeho Ahn;Chun-Sik Bae
    • Applied Microscopy
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    • v.50
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    • pp.29.1-29.6
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    • 2020
  • Erythronium japonicum (E. japonicum) and Corylopsis coreana Uyeki (C. coreana Uyeki, Korean winter hazel) have been shown to significantly decrease 1,3-dichloro-2-propanol (1,3-DCP)-induced generation of reactive oxygen species and CYP2E1 activity in HuH7, human hepatocytes. In this study, we expanded upon the previous study and investigated the effects of E. japonicum and C. coreana Uyeki extracts on 1,3-DCP-induced liver damage in rats. The pre-treatment of rats with these extracts alleviated a decrease in body weight and reduced 1,3-DCP-induced increase in catalytic activities of hepatic enzymes, such as aspartate aminotransferase and alanine aminotransferase, in the serum. Moreover, treatment with the extracts restored the 1,3-DCP-induced decreases in anti-oxidant enzyme activities, such as the activities of superoxide dismutase and catalase, in the rat liver. Histopathological studies also strongly supported the results of enzyme activities. These results suggest a possibility that the extracts of E. japonicum and C. coreana Uyeki can be a remedy for alleviating 1,3-DCP-induced liver damage in animals.

Effect of Dietary Fish Oil on Lipid Peroxidation in Rats Liver and Brain During Postnatal Development (어유섭취가 출생후 발달과정의 흰 쥐의 간과 뇌조직의 지질과산화와 그 관련기능에 미치는 영향)

  • 박명희
    • Journal of Nutrition and Health
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    • v.20 no.2
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    • pp.111-121
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    • 1987
  • Lipie peroxide formation, antiperoxidative s system and body adaptability for handling lipid p peroxide were examined in the first and second g generations of rats fed fish oil. Mackerel oil(MO) was used and four other dietary oils and fat, i.e. soybean oil(SO), perilla oil(PO), rapeseed oil(RO) and beef tallow(BT) were also employed to compare the effect of fish oil. Synthetic diets containing these five dietary fats at the level of 1O%(w/w), were given to the correspond­m ing groups of male and female rats weighing about 70 grams. After 34 days of feeding, male a and female rats were mated and their offsprings were raised throughout suckling (17, 26 days) and weanling (39 days) periods. Liver lipid pero­x xide level was highest in MO group of both first (mother rats after lactation) and second genera­t tions of 17 and 26 days old, but not of 39 days old. During suckling period, liver lipid peroxide level was well matched to total unsaturation of dietary fat. Brain lipid peroxide levels were not different among five groups. Liver $alpha$-tocopherol a and reduced glutathione (GSH) levels were lowest in MO fed first generation. In second generation, $alpha$-tocopherol level was also low in MO group, although the effect was less pronoun­c ced, but GSH level was not different from other groups. Oxidized glutathione (GSSG) level did not consistently vary by change in dietary fat. Glutathione peroxidase activity increased as young rats grew up to 39 days. Superoxide d dismutase activity change was insignificant by a age, but was shown as lowest in MO group. At the age of 26 and 39 days, liver glutatione peroxidase activity was increased as was level of lipid peroxide, suggesting that this is the one of the mechanisms responsible for body adapta­b bility for protection against the accumulation of lipid peroxide.

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Formation of Superoxide Anion in the Autoxidation of L-Ascorbic Acid in the Presence of Heavy Metal Ions (중금속 이온 존재하에서의 아스코르빈산 자동산화 과정에서 $O_2\bar{{\bullet}}$ 생성)

  • Kim, Mi-Ok
    • Korean Journal of Food Science and Technology
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    • v.33 no.3
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    • pp.378-383
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    • 2001
  • Formation of superoxide anion $O_2\bar{{\bullet}}$ in the autoxidation of L-ascorbic acid (AsA) in the presence of heavy metal ions were determined. The generation of $O_2\bar{{\bullet}}$ was studied by using superoxide dismutase (SOD) in aqueous and buffer solution, and using nitro bule tetrazolium (NBT) in methanol solution. The remaining amount of AsA was significantly higher in the presence of SOD than in its absence. It suggested that SOD stabilizes AsA in aqueous and buffer solution because of scavenging $O_2\bar{{\bullet}}$ formed during the autoxidation reaction of AsA in the presence of heavy metal ions. NBT has an absorption maximum at about 560 nm in methanol solution. The absorbance at 560 nm increased during the oxidation of AsA, suggested the formation of $O_2\bar{{\bullet}}$in methanol solution. Thus, the formation of $O_2\bar{{\bullet}}$ was confirmed during the autoxidation of AsA not only in aqueous solution but also in methanol solution in the presence heavy metal ions.

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Anti-oxidative and Anti-inflammatory Effects of Draconis Resina in Mouse Macrophage Cells (마우스 대식세포에서 혈갈(血竭)(Draconis Resina)의 항산화 및 항염증 효과)

  • Yi, Hyo-Seung;Heo, Sook-Kyoung;Yun, Hyun-Jeong;Choi, Jae-Woo;Jung, Jae-Ha;Park, Sun-Dong
    • The Korea Journal of Herbology
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    • v.23 no.2
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    • pp.179-192
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    • 2008
  • Objectives : Draconis Resina (DR) has been used as a traditional Korean herbal medicine since ancient times, and today it is used as a medication for wounds, tumors, diarrhea, rheumatism, in the itching of insect bites and with other conditions in the folk medicine. The aim of this study was to determine whether fractionated extracts of DR inhibit free radical generation, intracellular oxidation, production of nitrite, an index of NO, PGE2, iNOS, COX-2 and proinflammatory cytokines in lipopolysaccharide (LPS)-treated RAW 264.7 macrophages, Methods : DR extract prepared with methanol, and then fractionated with hexane, dichloromethane, ethylacetate, n-butanol and water. Inhibitory effect of DR onto free radical generation was determined by measuring DPPH, superoxide anions and nitric oxide scavenging activities in vitro. Cytotoxic activity of extracts on RAW 264.7 cells was measured using 5-(3-caroboxymeth-oxyphenyl)-2H-tetra-zolium inner salt (MTS) assay. Intracelluar oxidation was analysed by DCF-DA assay. The nitric oxide (NO) production was measured by Griess reagent system. The levels of iNOS and COX-2 expression were confirmed by western blot. And proinflammatory cytokines were measured by ELISA kit. Results : Our results indicated that fractionated extracts, especially dichloromethane and ethyl acetate extracts, significantly inhibited free radical generation, the LPS-induced H202, NO, PGE2 production and iNOS, COX-2 expression accompanied by an attenuation of TNF-${\alpha}$, IL-$1{\beta}$ and IL-6 formation in macrophages. Conclusions : Our results indicate that dichloromethane and ethyl acetate extracts of DR have potential as an agent of chronic inflammatory diseases.

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Phototoxicity Evaluation of Pharmaceutical Substances with a Reactive Oxygen Species Assay Using Ultraviolet A

  • Lee, Yong Sun;Yi, Jung-Sun;Lim, Hye Rim;Kim, Tae Sung;Ahn, Il Young;Ko, Kyungyuk;Kim, JooHwan;Park, Hye-Kyung;Sohn, Soo Jung;Lee, Jong Kwon
    • Toxicological Research
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    • v.33 no.1
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    • pp.43-48
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    • 2017
  • With ultraviolet and visible light exposure, some pharmaceutical substances applied systemically or topically may cause phototoxic skin irritation. The major factor in phototoxicity is the generation of reactive oxygen species (ROS) such as singlet oxygen and superoxide anion that cause oxidative damage to DNA, lipids and proteins. Thus, measuring the generation of ROS can predict the phototoxic potential of a given substance indirectly. For this reason, a standard ROS assay (ROS assay) was developed and validated and provides an alternative method for phototoxicity evaluation. However, negative substances are over-predicted by the assay. Except for ultraviolet A (UVA), other UV ranges are not a major factor in causing phototoxicity and may lead to incorrect labeling of some non-phototoxic substances as being phototoxic in the ROS assay when using a solar simulator. A UVA stimulator is also widely used to evaluate phototoxicity in various test substances. Consequently, we identified the applicability of a UVA simulator to the ROS assay for photoreactivity. In this study, we tested 60 pharmaceutical substances including 50 phototoxins and 10 non-phototoxins to predict their phototoxic potential via the ROS assay with a UVA simulator. Following the ROS protocol, all test substances were dissolved in dimethyl sulfoxide or sodium phosphate buffer. The final concentration of the test solutions in the reaction mixture was 20 to $200{\mu}M$. The exposure was with $2.0{\sim}2.2mW/cm^2$ irradiance and optimization for a relevant dose of UVA was performed. The generation of ROS was compared before and after UVA exposure and was measured by a microplate spectrophotometer. Sensitivity and specificity values were 85.7% and 100.0% respectively, and the accuracy was 88.1%. From this analysis, the ROS assay with a UVA simulator is suitable for testing the photoreactivity and estimating the phototoxic potential of various test pharmaceutical substances.

Protective Effects of Portulaca oleracea L. Extract against Matrix Metalloproteinase Production and Reactive Oxygen Species Generation Induced by Ultraviolet B Radiation in Human Keratinocytes (쇠비름 추출물의 UVB 자외선 조사에 의한 인간각질형성세포 손상에 대한 보호 효과)

  • Oh, Jung Hwan;Karadeniz, Fatih;Lee, Jung Im;Park, So Young;Seo, Youngwan;Kong, Chang-Suk
    • Journal of Life Science
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    • v.28 no.8
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    • pp.892-899
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    • 2018
  • Portulaca oleracea L. is an edible plant widely consumed in daily diet throughout Europe, Asia and America. In this study, protective effects of P. oleracea L. extracts against oxidative stress and matrix metalloproteinase (MMP) activity induced by ultraviolet B (UVB) radiation were investigated using HaCaT immortal human keratinocytes. In this context, the mRNA and protein productions of MMPs (MMP-1, -2, and -9) and type I procollagen, which are major markers of photoaging induced by UVB radiation in HaCaT keratinocytes, were evaluated. Furthermore, UVB-induced reactive oxygen species (ROS) generation and mRNA and protein expression levels of superoxide dismutase-1 (SOD-1), oxygenase-1 (OH-1), and nuclear factor-erythroid 2-related factor-2 (Nrf-2), all of which are associated with the antioxidant balance, were investigated. As shown by the results, UVB radiation induced ROS formation and led to increased production of MMPs and decreased collagen production in human keratinocytes, which resulted in skin photoaging or photodamage. The treatment with P. oleracea L. extracts downregulated MMP (MMP-1, -2, and -9) production and upregulated type I procollagen expression in UVB-induced HaCaT cells. Furthermore, treatment with the extracts decreased UVB-induced ROS generation and increased the expression of antioxidant enzymes, such as SOD-1 and OH-1, through the Nrf-2 pathway. Taken together, these results suggest that P. oleracea L. extracts could be a potential cosmeceutical agent for the prevention of skin photoaging or photodamage.

Effects of Nitrofurantoin on Lipid Peroxidation and Reactive Oxygen Radical Generation in Porcine Lung Microsome (Nitrofurantion이 폐장 미크로솜 지질과산화와 반응성 산소 라디칼 생성에 미치는 영향)

  • Paick, Jae-Seung;Kim, Si-Whang;Kim, Hae-Won;Chung, Myung-Hee;Kim, Myung-Suk
    • The Korean Journal of Pharmacology
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    • v.21 no.1
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    • pp.34-48
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    • 1985
  • In vitro effects of nitrofurantoin, an antimicrobial agent for acute and chronic urinary tract infection, on the lung microsomal lipid peroxidation and the generation of reactive oxygen radicals were investigated to elucidate the biochemical mechanisms of its in vivopulmonary toxicity. The interaction of nitrofurantoin with porcine lung microsome resulted in significant lipid peroxidation. In addition, nitrofurantoin stimulated the generation of reactive oxygen radicals, $O^{-}_{2}{\cdot},\;H_2O_2$ as well as a highly reactive secondary oxygen species, $OH{\cdot}$. The stimulation of lipid peroxidation was inhibited not only by superoxide dismutase and catalase, but also by hydroxyl radical scavengers, mannitol and thiourea. Neither singlet oxygen $({^1}O_{2})$ was detected during the incubation of microsome with nitrofurantoin, nor lipid peroxidation was inhibited by singlet oxygen scavengers. When incubated anaerobically under the nitrogen atmosphere, the ability of nitrofurantoin to stimulatle lipid peroxidation was abolished. It appears that NADPH-dependent metaboliam of nitrofurantoin in pulmonary microsome under aerobic condition is accompanied by the stimulation of lipid peroxidation through the mediation of reactive oxygen radicals, particularly hydroxyl radical. It is strongly suggested from these results that the stimulation of pulmonary microsomal lipid peroxidation by the reactive oxygen radical may be a in vivo mechanism of pulmonary toxicity caused by nitrofurantoin.

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Effects of Sotosaja-hwan on the Generation of ROS, RNS, and on the Expression of NF-${\kappa}B$-dependent Proteins in ob/ob Mouse (소도사자환이 ob/ob mouse에서 ROS/ RNS 생성 억제 및 NF-${\kappa}B$ 의존성 단백질에 미치는 영향)

  • Bang, Yong-Suk;Jeong, Ji-Cheon
    • The Journal of Korean Medicine
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    • v.30 no.1
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    • pp.51-63
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    • 2009
  • Objectives: Peroxynitrite ($ONOO^-$), superoxide anion radical (${\cdot}{O_2}^-$ and nitric oxide (NO) are cytotoxic because they can oxidize several cellular components such as proteins, lipids and DNA. They have been implicated in the aging processes, and age-related diseases such as Alzheimer's disease, rheumatoid arthritis, cancer, diabetes, obesity and atherosclerosis. The aim of this study was to investigate the $ONOO^-$, NO, ${\cdot}{O_2}^-$ scavenging and NF-${\kappa}B$ related anti-inflammatory activities of Sotosaja-hwan in ob/ob mice. Methods: Mice were grouped and treated for 5 weeks as follows. Both the normal lean (C57/BL6J black mice) and control obese (ob/ob mice) groups have 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. For this study, the fluorescent probes, namely 2',7'-dichlorodihydrofluorescein diacetate (DCFDA), 4,5-diaminofluorescein (DAF-2) and dihydrorhodamine 123 (DHR 123) were used. Western blotting was performed using anti-phospho-$I{\kappa}B$-${\alpha}$, anti-IKK-${\alpha}$, anti-NF-${\kappa}B$ (p50, p65), anti-COX-2, anti-iNOS, anti-YCAM-1 and anti-MMP-9 antibodies, respectively. Results: Sotosaja-hwan inhibited the generation of $ONOO^-$, NO and ${\cdot}{O_2}^-$ in the lipopolysaccharide (LPS)-treated mouse kidney postmitochondrial fraction in vitro. The generation of $ONOO^-$, NO, ${\cdot}{O_2}^-$ and PGE2 were inhibited in the Sotosaja-hwan-administered ob/ob mice groups. The GSH/GSSG ratio was decreased in the ob/ob mice, whereas the ratio was improved in the Sotosaja-hwan-administered groups. Sotosaja-hwan inhibited the protein expression levels of phospho-$I{\kappa}B$-${\alpha}$, IKK-${\alpha}$, NF-${\kappa}B$ (p50, p65), COX-2, iNOS, YCAM-1 and MMP-9 genes. Conclusions: These results suggest that Sotosaja-hwan is an effective $ONOO^-$, ${\cdot}{O_2}^-$ and NO scavenger and has NF-kB related anti-inflammatory activity in ob/ob mice. Therefore, Sotosaja-hwan might be a potential therapeutic drug against the inflammation process and inflammation-related diseases.

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