• Title/Summary/Keyword: Superoxide assay

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The Effect of Metallothionein on the Activity of Enzymes Invelved in Remival of Reactive Oxygen Species

  • Go, Mun Ju;Kim, Hui Jeong
    • Bulletin of the Korean Chemical Society
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    • v.22 no.4
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    • pp.362-366
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    • 2001
  • To show the effects of metallothionein (MT) on the activity of enzymes involved in the removal of reactive oxygen species, MT has been added to the assay systems of superoxide dismutase (SOD), catalase and peroxidase. We have used assay systems of SOD based on NADPH oxidation and nitrite formation from hydroxylammonium chloride as an assay of superoxide breakdown rate. The two assay systems showed different results at the high concentration of MT. MT showed the scavenging of superoxide in the SOD assay system in the presence and absence of SOD. MT added to the SOD assay system behaved as an activator of SOD, but apo-MT behaved as an inhibitor. When MT was added to the assay system in the presence of a fixed amount of SOD, the breakdown rate of superoxide increased. The effects of MT on the decomposition of hydrogen peroxide and the activity of catalase and peroxidase decomposing hydrogen peroxide were evaluated. MT decreased the activities of catalase and peroxidase. We have concluded that the function of MT as an antioxidant might effect the level of superoxide scavenging and not the level of hydrogen peroxide.

Antioxidative Constituents from Lycopus lucidus

  • Woo, Eun-Rhan;Piao, Mei-Shan
    • Archives of Pharmacal Research
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    • v.27 no.2
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    • pp.173-176
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    • 2004
  • Three phenolic compounds, rosmarinic acid (1), methyl rosmarinate (2), ethyl rosmarinate (3), and two flavonoids, luteolin (4), luteolin-7-O-$\beta$-D-glucuronide methyl ester (5) were isolated from the aerial part of Lycopus lucidus (Labiatae). Their structures were determined by chemical and spectral analysis. Compounds 1-5 exhibited potent antioxidative activity on the NBT superoxide scavenging assay. The $IC_{50}$ values for compounds 1-5 were 2.59, 1.42, 0.78, 2.83, and 3.05 $\mu\textrm{g}$/mL respectively. In addition, five compounds were isolated from this plant for the first time.

An Improved method in Screening of Superoxide and Hydroxyl Radical Scavenging Activities of Plant Medicinal Extracts (생약 추출물에 의한 superoxide와 hydroxyl 라디칼 소거능 검색 방법의 개선)

  • Lee, Ho-Sub;Kang, Dae-Gill
    • Korean Journal of Pharmacognosy
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    • v.32 no.3 s.126
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    • pp.253-256
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    • 2001
  • The present study was designed for the improvement of routine measurement of superoxide and hydroxyl radical scavenging activities utilized by a microplate reader. Superoxide radical scavenging activity by the ascorbic acid, which is a well-known superoxide scavenger, was determined in a dose-dependent manner. Hydroxyl radical scavenging activity by the thiourea, which is a well-known hydroxyl radical scavenger, was also well detected in a dose-dependent manner. Our results suggest that the use of microplate reader to assay the superoxide and hydroxyl radical scavenging activities improves the accuracy of data and enables the use of much smaller amounts of samples and/or reagents, with much simpler experimental procedure. Therefore, These methods appear to be suitable for screening of superoxide and hydroxyl radical scavenging activities in both the plant medicinal extracts and the isolated compounds.

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Evaluation of in vitro antioxidant activities of the methanol extracts of Glinus oppositifolius and Trianthema decandra

  • Mazumder, Upal Kanti;Gupta, Malaya;Haldar, Pallab Kanti;Kandar, Chandi Charan
    • Advances in Traditional Medicine
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    • v.7 no.3
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    • pp.290-296
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    • 2007
  • Glinus oppositifolius and Trianthema decandra belonging to the Ficoidaceae family were commonly used by tribal peoples for the treatment of liver disorders and cancer. The preliminary phytochemical screening of those plants showed the presence of flavonoids, terpenoids, tannins and saponins. The aim of this study was to evaluate the in vitro antioxidant activities of the methanol extracts of Glinus oppositifolius (MEGO) and Trianthema decandra (METD). The antioxidative capacities of MEGO and METD were determined by the following four complementary assay; DPPH radical scavenging assay, superoxide anion generation by xanthine-xanthine Oxidase assay, hydroxyl radical scavenging assay and $Fe^{2+}$-ascorbate induced by lipid peroxidation assay. The $IC_{50}$ values of the both extracts were calculated from the inhibition curve. The $IC_{50}$ MEGO and METD in DPPH, superoxide anion, hydroxyl radical scavenging and lipid peroxidation assay are 1.85, 7.31, 13.95, 22.82 and 2.21, 9.78, 14.87, 19.76 ${\mu}g/ml$ respectively. Both the extracts exhibited a significant antioxidant effects.

The Detection of Superoxide Dismutase Activity and Isozyme Pattern of Panax ginseng C.A. Meyer Leaves (인삼엽에서 Superoxide Dismutase Activity 측정 및 Isozyme Pattern 검정)

  • Yang, Deok-Jo;Kim, Myeong-Sik;Lee, Seong-Jong
    • Journal of Ginseng Research
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    • v.11 no.1
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    • pp.24-31
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    • 1987
  • We studied a assay method on the measurement of superoxide dismutase (SOD Superoxide : superoxide oxidoreductase, EC. 1. 15. 1. 1) activity with photoreduced flavin and nitroblue tetrazolium (NBT) as superoxide (${O_2}^{-}$) source and detector, respectively. The $\Delta$E (1000 ng SOD$.$$min.)^{-1}$ of photoreduced flavin-NBT system was 0.08, whereas that of xanthine-xanthine-cytochrome system used broadly in experiments was 0.014. Therefore, the new method was regarded more simple and utilizable than xanthine-xanthine cytochrome system method. In the present paper, we also carried out to investigate the SOD activity and isozyme pattern for the parpose of study of leaf-burning disease in ginseng (Panax ginseng C.A. Meyer) leaves.

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Antioxidative Activity of Water Extract of Different Parts of Acanthopanax divaricatus var, albeofructus (흰털오가피 부위별 물추출물의 항산화활성)

  • Lyu, Su-Yun;Kim, Ji-Young;Noh, Bin-Na;Park, Won-Bong
    • YAKHAK HOEJI
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    • v.50 no.3
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    • pp.191-198
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    • 2006
  • Acanthopanax species have traditionally been used as a tonic, a sedative as well as in the treatment of rheumatism, hypertension and diabetes. In the present study, oxidative stress was induced in Vero cells by incubating the cells with glucose and the cell viability was measured by MTT assay. The concentration of glucose which 50% of cell viability was 125 mM $(IC_{50})$ and the cell viability was increased to $87.6{\pm}8.8%$ by treatment of the extracts of Acanthopanax divaricatus var. albeofructus. The antioxidative activity of water extract of different parts of the Acanthopanax plant was investigated by DPPH (1,1-diphenyl-2-picrylhydrazyl) assay, xylenol orange assay, TBARS (thiobarbituric acid reactive substances) assay and enzyme (superoxide anion and catalase) assay. Each extract (leaves, root, stem and fruits) of the plant showed free radical and $H_2O_2$ scavenging activity. The extract also inhibited lipid peroxidation and recovered enzyme (superoxide anion dismutase and catalase) activity in Vero cells treated with glucose.

Validation of Analytical Methods for Plasma Total Antioxidant Capacity by Comparing with Urinary 8-Isoprostane Level

  • Lee, Sang Gil;Wang, Taoran;Vance, Terrence M.;Hurbert, Patrice;Kim, Dae-Ok;Koo, Sung I.;Chun, Ock K.
    • Journal of Microbiology and Biotechnology
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    • v.27 no.2
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    • pp.388-394
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    • 2017
  • Although several analytical methods for measuring total antioxidant capacity (TAC) have been applied to biological samples, there were often dissimilar results due to the different principles of methods applied. Thus, this study aimed to validate four conventional analytical methods for measuring plasma TAC, including the ABTS assay, DPPH assay, FRAP assay, and ORAC assay, by comparing with urinary 8-isoprostane concentration. In addition, TAC results were compared with antioxidant enzyme activities including superoxide dismutase (SOD) and glutathione peroxidase in erythrocyte, and catalase in plasma. Plasma TAC measure by ABTS assay was strongly correlated with the result by FRAP assay. Plasma TAC by FRAP and ORAC assays were negatively correlated with erythrocyte SOD activity. The agreement among the four TAC assay methods and 8-isoprostane was determined using 95% prediction limits of linear regression, expressed as the mean of 8-isoprostane ${\pm}95%$ prediction limits. The ABTS method better agreed with 8-isoprostane than the other methods, demonstrating narrow prediction of limits. Furthermore, only plasma TAC determined by the ABTS assay was inversely correlated with urinary 8-isoprostane (r = -0.35, p < 0.05). In summary, the ABTS assay would be an appropriate method to measure overall plasma antioxidant capacity and predict the body's antioxidant status.

Antioxidant Activities and Whitening Effect from Lindera obtusiloba BL. Extract (생강나무 추출물의 항산화 활성과 미백효과)

  • Bang, Chae-Young;Won, Eun-Kyung;Park, Kuen-Woo;Lee, Gwang-Won;Choung, Se-Young
    • YAKHAK HOEJI
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    • v.52 no.5
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    • pp.355-360
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    • 2008
  • In this study we investigated antioxidant activity of against several free radicals and skin whitening effect of 70% ethanol extract (leaf extracts and branch/stem mixed) of Lindera obtusiloba BL. Antioxidant activity was assessed by DPPH, superoxide radical and hydroxyl radical assays. The Lindera obtusiloba BL. extract had antioxidant activity dose dependently with an ${IC}_{50}$ value of 243.14 and 181.10 ${\mu}g$/ml for DPPH, 165.77 and >1500 ${\mu}g$/ml for non-enzymatic system of superoxide radical assay, 35.47 and >100 ${\mu}g$/ml for enzymatic system of superoxide radical assay, 1.21 mg/ml for hydroxyl radical assay. In addition we tested tyrosinase inhibition activity and melanin contents on B16 melanoma F10. B16 melanoma cell was treated by such sample as 1, 5, 10 and 50 ${\mu}g$/ml for 72 hr and tyrosinase inhibition was tested. Melanogenesis was inhibited to 22% at the dose of 50 ${\mu}g$/ml and tyrosinase was inhibited to 45.2% at the same dose. In conclusion Lindera obtusiloba BL had potent antioxidant activity and inhibitory activity of tyrosinase and melanin formation. It could be developed as the health functional food and functional cosmetic resources.

Antioxidant Property of the Gagam-Hyungbang-Gihwang-tang Using Biochemical Markers of Carcinogenesis (가감형방지황탕 열수 추출물이 항산화 작용에 미치는 영향)

  • Han Jin-Soo;Park Seong-Sik
    • The Journal of Korean Medicine
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    • v.26 no.3 s.63
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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.

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Antioxidant Effects of Solvent Fraction from Sanguisorbae officinalis L. with Acetone (오이풀 아세톤 추출물을 이용한 용매 분획물의 항산화 효과)

  • Kim, Hui-Yeong;Yeo, Shin-Il;Lee, Jin-Tae
    • Journal of Applied Biological Chemistry
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    • v.54 no.2
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    • pp.89-93
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    • 2011
  • The solvent extracts of Sanguisorbae officinalis L. were investigated for the activities of antioxidants as a functional ingredient for cosmetic products. The electron donating effect of ethyl acetate layer and n-butyl alcohol layer was appeared similar activity with positive control butylated hydroxy anisole (BHA) at all concentrations. In addition, in 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS) radical scavenging assay, ethyl acetate layer, n-butyl alcohol layer and water layer were over 99% effect at all concentrations and higher than that of BHA. Also in hydrogen peroxide scavenging assay, ethyl acetate layer and n-butyl alcohol layer were higher than that of positive control ascorbic acid. The measured superoxide dismutase (SOD)-like activity of n-butyl alcohol was more than 50% at concentration of 1,000 ${\mu}g/mL$ and superoxide anion radical scavenging ability showed more than 45% at 1,000 ${\mu}g/mL$ of n-butyl alcohol layer. All these findings suggested that ethyl acetate layer and n-butyl alcohol layer have a great potential as a cosmeceutical ingredient with an antioxidant effect.