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1.
《Bioscience, biotechnology, and biochemistry》2013,77(6):1324-1331
Phosvitin, a phosphoprotein known as an iron-carrier in egg yolk, binds almost all the yolk iron. In this study, we investigated the effect of phosvitin on Fe(II)-catalyzed hydroxyl radical (?OH) formation from H2O2 in the Fenton reaction system. Using electron spin resonance (ESR) with 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) and deoxyribose degradation assays, we observed by both assays that phosvitin more effectively inhibited ?OH formation than iron-binding proteins such as ferritin and transferrin. The effectiveness of phosvitin was related to the iron concentration, indicating that phosvitin acts as an antioxidant by chelating iron ions. Phosvitin accelerates Fe(II) autoxidation and thus decreases the availability of Fe(II) for participation in the ?OH-generating Fenton reaction. Furthermore, using the plasmid DNA strand breakage assay, phosvitin protected DNA against oxidative damage induced by Fe(II) and H2O2. These results provide insight into the mechanism of protection of the developing embryo against iron-dependent oxidative damage in ovo. 相似文献
2.
比色法测定Fenton反应产生的羟自由基 总被引:107,自引:0,他引:107
Fenton反应产生的羟自由基能与水杨酸生成羟基化产物2,3-二羟基苯甲酸,用比色法测定其含量能间接测定羟自由基的生成量.通过对测定条件的研究,得到最佳的测定方案.可作为一种简便的筛选羟自由基清除剂的方法 相似文献
3.
比色法测定Fenton反应产生的羟自由基及其应用 总被引:46,自引:0,他引:46
Fenton反应产生的羟自由基与二甲亚砜反应,生成甲基亚磺酸,再与坚牢蓝BB盐反应生成偶氮砜,比色法测定其含量可间接测定OH·的生成量. 通过对测定条件的研究,得到最佳实验方案. 抗氧化剂药物硫脲和抗坏血酸与羟自由基清除率具有明显的量效关系. 测定了核桃、黑芝麻等几种天然食物的水提取物清除羟自由基的功能. 此法可用于羟自由基清除剂的筛选. 相似文献
4.
Eric Lancelot Marie-Louise Revaud Roger G Boulu Michel Plotkine Jacques Callebert 《Free radical biology & medicine》1997,23(7):1031-1034
Various in vitro experiments have indicated that oxygen-derived free radicals may contribute to excitotoxic neuronal death. In the present study we induced excitotoxicity in rat striatum by perfusing glutamate at a high concentration through a microdialysis probe. We observed an increased formation of hydroxyl radicals (˙OH) during the perfusion of the excitotoxin and an extensive striatal lesion 24 h after the insult. The spin trap, -phenyl-N-tert-butylnitrone (PBN), attenuated both hydroxyl radical levels and the volume of the lesion. This result suggests that the neuroprotection may be due to a free radical scavenging mechanism. It also implies that PBN may be used in pathological situations involving excitotoxicity such as stroke, brain trauma, and chronic neurologic diseases. 相似文献
5.
《Bioscience, biotechnology, and biochemistry》2013,77(9):1507-1509
Ansamycin antibiotics (1–4) were isolated from the cultured broth of Streptomyces sp. USF-319 strain as a result of our screening for free radical scavengers. They inhibited the bactericidal effect of the Fenton reagent toward Bacillus subtilis by their radical scavenging activity. Some of them also showed inhibitory activity against lipid peroxidation and lipoxygenases. 相似文献
6.
Jonathan S. B. Park Paul M. Wood Michael J. Davies Bruce C. Gilbert Adrian C. Whitwood 《Free radical research》1997,27(5):447-458
The reaction of FeII oxalate with hydrogen peroxide and dioxygen was studed for oxalate concentrations up to 20 mM and pH 2-5, under which conditions mono- and bis-oxalate comlexes (FeII(ox) and FeII(ox)22-) and uncomplexed Fe2+ must be considered. The reaction of FeII oxalate with hydrogen peroxide (Fe2+ + H2O2 → Fe3+ + *OH + OH-) was monitored in continuous flow by ESR with t-butanol as a radical trap. The reaction is much faster than for uncomplexed Fe2+ and a rate constant, k = 1 × 104 M-1 s-1 is deduced for FeII(ox). The reaction of FeII oxalate with dioxygen is strongly pH dependent in a manner which indicates that the reactive species is FeII(ox)22-, for which an apparent second order rate constant, k = 3.6 M-1 s-1, is deduced. Taken together, these results provide a mechanism for hydroxyl radical production in aqueous systems containing FeII complexed by oxalate. Further ESR studies with DMPO as spin trap reveal that reaction of FeII oxalate with hydrogen peroxide can also lead to formation of the carboxylate radical anion (CO2*-), an assignment confirmed by photolysis of FeIII oxalate in the presence of DMPO. 相似文献
7.
Anthony J. Nappi Emily Vass 《Biochimica et Biophysica Acta (BBA)/General Subjects》1998,1425(1):159-167
The differing effects of O-methylated catecholamines and their dihydroxyphenyl precursors on the production of ?OH were quantified using a previously established specific salicylate hydroxylation assay in conjunction with a sensitive electrochemical detection system. The production of ?OH by the Fenton reaction was diminished significantly by O-methylated catecholamines (O-methyldopa, O-methyldopamine, O-methyltyrosine, and N-acetyl-O-methyldopamine), whereas radical production was augmented by dihydroxyphenyls (DOPA, dopamine, and N-acetyldopamine), including those with methylated side chains (N-methyldopamine and α-methyldopa). Monohydroxyphenyls such as octopamine, tyramine, tyrosine, and α-methyltyrosine had little or no effect on radical production. These data show that a methyl group positioned on the side chain of a catecholamine does not alter its pro-oxidant behavior, while a methyl group positioned on the aromatic ring renders the catecholamine sterically or kinetically unfavorable for coordination with transition metals, thus preventing the promotion of Fenton chemistry. These results highlight the importance of O-methylation in forming catechols that are less reactive than their dihydroxyphenyl precursors. Thus, factors regulating the methylation of brain catecholamines may play a crucial role in mediating neuronal integrity during aging and in the pathogenesis of certain neurodegenerative disorders. Competitive side-chain methylation reactions may sustain or perpetuate some dihydroxyphenyls, creating an oxidatively less favorable environment for cells than would result from compounds formed by O-methylation. 相似文献
8.
《Bioscience, biotechnology, and biochemistry》2013,77(12):3014-3018
To clarify the effect of superoxide dismutase (SOD) on the formation of hydroxyl radical in a standard reaction mixture containing 15 μM of xanthone, 0.1 M of 5,5-dimethyl-1-pyrroline N-oxide (DMPO), and 45 mM of phosphate buffer (pH 7.4) under UVA irradiation, electron paramagnetic resonance (EPR) measurements were performed. SOD enhanced the formation of hydroxyl radicals. The formation of hydroxyl radicals was inhibited on the addition of catalase. The rate of hydroxyl radical formation also slowed down under a reduced oxygen concentration, whereas it was stimulated by disodium ethylenediaminetetraacetate (EDTA) and diethyleneaminepentaacetic acid (DETAPAC). Above findings suggest that O2, H2O2, and iron ions participate in the reaction. SOD possibly enhances the formation of the hydroxyl radical in reaction mixtures of photosensitizers that can produce O2 ?·. 相似文献
9.
Mark J. Burkitt 《Free radical research》1993,18(1):43-57
Several investigators have challenged the widely held view that the hydroxyl radical is the primary oxidant formed in the reaction between the ferrous ion and hydrogen peroxide. In recent studies, using the ESR spin trapping technique, Yamazaki and Piette found that the stoichiometry of oxidant formation in the reaction between Fe2+ and H2O2 often shows a marked deviation from the expected value of 1:1 (I. Yamazaki and L. H. Piette (1990) J. Am. Chem. Soc. 113, 7588-7593). In order to account for these observations, it was suggested that additional oxidizing species are formed, such as the ferryl ion (FeO2+), particularly when iron is present at high concentration and chelated to EDTA.
In this paper it is shown that secondary reactions, involving the redox cycling of iron and the oxidation of the hydroxyl radical adduct of the spin trap 5,5-dimethyl-1-pyrroline-N-oxide(DMPO) by iron, operate under the reaction conditions employed by Yamazaki and Piette. Consequently, the stoichiometry of oxidant formation can be rationalized without the need to envisage the formation of oxidizing species other than the hydroxyl radical. It is also demonstrated that the iron(III) complex of DETAPAC can react directly with DMPO to form the DMPO hydroxyl radical adduct (DMPO/OH) in the absence of hydrogen peroxide. Therefore, to avoid the formation of (DMPO/OH) as an artefact, it is suggested that DETAPAC should not be used as a reagent to inactivate containating adventitious iron in experiments using DMPO. 相似文献
In this paper it is shown that secondary reactions, involving the redox cycling of iron and the oxidation of the hydroxyl radical adduct of the spin trap 5,5-dimethyl-1-pyrroline-N-oxide(DMPO) by iron, operate under the reaction conditions employed by Yamazaki and Piette. Consequently, the stoichiometry of oxidant formation can be rationalized without the need to envisage the formation of oxidizing species other than the hydroxyl radical. It is also demonstrated that the iron(III) complex of DETAPAC can react directly with DMPO to form the DMPO hydroxyl radical adduct (DMPO/OH) in the absence of hydrogen peroxide. Therefore, to avoid the formation of (DMPO/OH) as an artefact, it is suggested that DETAPAC should not be used as a reagent to inactivate containating adventitious iron in experiments using DMPO. 相似文献
10.
The technique of E.S.R. spectroscopy, when employed in conjunction with a continuous flow system, provides direct evidence for the nature of free radicals formed from organic substrates in the presence of FeII and H2O2 in aqueous solution. It is shown, both via the identification of hydroxyl-radical adducts to alkenes and via the observed site-selectivity of radical attack, that the hydroxyl radical is formed as the reactive intermediate in the presence of various chelators (e.g. EDTA, DTPA). This approach also allows the rate constants for the FeII-H2O2 reaction in the presence of the different chelates to be determined; values obtained are in reasonable agreement with most of those measured by other methods. Examples of radical oxidation (by FeIII) and reduction (by FeII) are revealed. 相似文献
11.
The polymorphonuclear leukocyte secretes both O2-and H2O2 when stimulated by various soluble or particulate stimuli. Since a rcaction involving iron, O2-, and H2O2 could generate the hydroxyl radical (HO.) there has been speculation that the HO-may participate in the bactericidal activity of the neutroph-il. A variety of water-soluble HO. scavengers have been used to test for the participation of HO. and the results imply that HO. might participate. However, other workers have not been able to detect the formation of significant amounts of HO-by the activated neutrophil. We have examined the effect of several commonly used HO. radical scavengers on the ability of the neutrophil to secrete O2-and H2O2. Several of these compounds actively inhibit secretion without affecting the viability of the neutrophil. After considering the various complications inherent in using water soluble radical scavengers, we suggest that they only be used with well defined experimental systems. 相似文献
12.
13.
Using ESR with 5,5-dimethyl-l-pyrroline N-oxide (DMPO) as a spin-trapping reagent, we measured the levels of free radical species generated from living cells of Chlorella vulgaris var. vulgails (IAM C-534). To investigate the production of free radicals in the living Chlorella vulgaris cells, the influence of DMPO toward the intact cells of the Chlorella vulgaris using the O2 evolution rate was first studied as a guide. Since the 02 evolution rate was not changed by DMPO, it was judged that DMPO has no toxicity toward the intact cells of Chlorella vulgaris.
Only hydroxyl radicals (-OH) were detected as the DMPO-OH adduct in the suspension of intact cells of Chlorella vulgaris irradiated with visible light. Moreover, since production of -OH was inhibited by some hydroxyl radical scavengers such as KI and ethanol, production of -OH was proved to be due to hydroxyl radicals. It was also clear that the intensity of OH increased with increasing irradiation intensity of visible light. Therefore, it was suggested that -OH might be one of the photoinhibition factors of the intact Chlorella vulgaris cells in severe light conditions. 相似文献
Only hydroxyl radicals (-OH) were detected as the DMPO-OH adduct in the suspension of intact cells of Chlorella vulgaris irradiated with visible light. Moreover, since production of -OH was inhibited by some hydroxyl radical scavengers such as KI and ethanol, production of -OH was proved to be due to hydroxyl radicals. It was also clear that the intensity of OH increased with increasing irradiation intensity of visible light. Therefore, it was suggested that -OH might be one of the photoinhibition factors of the intact Chlorella vulgaris cells in severe light conditions. 相似文献
14.
壳聚糖季铵盐合成及其抗氧化性能研究 总被引:9,自引:1,他引:9
通过Eschweiler Clarke反应得到N,N-二甲基壳聚糖中间体。然后与碘甲烷反应生成水溶性的壳聚糖季铵盐:N,N,N-三甲基壳聚糖碘化铵(TMCI)。检测三种不同取代度的季铵盐产品对羟基自由基(.OH)、超氧自由基(O2-.)的清除率。结果发现:取代度为33.9%时,对.OH的最大清除率为40.2%,对O2-.的最大抑制率为65.6%。并且季铵盐产品对两种自由基的清除率都随着取代度的增大而减小,取代度为80.3%的产品对这两种自由基几乎无清除作用。 相似文献
15.
Hydroxyl Free Radical Adduct of Deoxyguanosine: Sensitive Detection and Mechanisms of Formation 总被引:38,自引:0,他引:38
Robert A. Floyd Julia J. Watson Peter K. Wong Dale H. Altmiller Robert C. Rickard 《Free radical research》1986,1(3):163-172
DNA or 2-deoxyguanosine reacts with hydroxyl free radical to form 8-hydroxy-deoxyguanosine (8-OH-dG). We found that 8-OH-dG can be effectively separated from deoxyguanosine by high pressure liquid chromatography and very sensitively detected using electrochemical detection. The sensitivity by electrochemical detection is about one-thousand fold enhanced over optical detection. Utilizing deoxyguanosine in bicarbonate buffer it was found that ferrous ion, but not ferric ion, was effective in forming 8-OH-dG. The hydroxyl free radical scavenging agents, thiourea and ethanol, were very effective in quenching Fe(11) mediated 8-OH-dG formation, but superoxide dismutase had very little effect. 相似文献
16.
《Free radical research》2013,47(3):163-172
DNA or 2-deoxyguanosine reacts with hydroxyl free radical to form 8-hydroxy-deoxyguanosine (8-OH-dG). We found that 8-OH-dG can be effectively separated from deoxyguanosine by high pressure liquid chromatography and very sensitively detected using electrochemical detection. The sensitivity by electrochemical detection is about one-thousand fold enhanced over optical detection. Utilizing deoxyguanosine in bicarbonate buffer it was found that ferrous ion, but not ferric ion, was effective in forming 8-OH-dG. The hydroxyl free radical scavenging agents, thiourea and ethanol, were very effective in quenching Fe(11) mediated 8-OH-dG formation, but superoxide dismutase had very little effect. 相似文献
17.
鸟嘌呤碱基与羟基自由基反应的密度泛函理论 总被引:3,自引:0,他引:3
羟基自由基 (·OH)进攻嘌呤碱基是破坏核酸造成DNA断链损伤的重要原因之一 .采用密度泛函 (DFT)理论中B3LYP方法在 6— 31G基组水平上对鸟嘌呤 (G)受羟基自由基进攻形成的各种可能产物自由基进行几何全优化 .根据总能量、键长和自旋密度的计算结果 ,从理论上确认了C 5和C 8位加成机制 .得产物自由基G5OH·、G8OH· ,且G5OH·易与N 11位H脱水得一个更稳定的产物自由基 ,而G8OH·不易发生开环反应 ,得到与实验一致的结论 .这些稳定自由基的形成造成DNA断链损伤 相似文献
18.
Thomas D. Tullius 《Free radical research》1991,13(1):521-529
The Fenton reaction of iron(II) EDTA with hydrgen peroxide, performed in the presence of ascorbateion. has proven to be useful as a probe of structure in DNA systems. Two aspects of this chemistry are discussed: the identity of the active DNA cleaving agent produced by this reagent, and the application of the Fenton reaction to the determination of the structure of the Holliday junction, the four-stranded DNA molecule that is a key intermediate in recombination. The cleavage pattern of the Holliday junction has pseudo-twofold symmetry, putting important constraints on possible structures. 相似文献
19.
《Free radical research》2013,47(1):521-529
The Fenton reaction of iron(II) EDTA with hydrgen peroxide, performed in the presence of ascorbateion. has proven to be useful as a probe of structure in DNA systems. Two aspects of this chemistry are discussed: the identity of the active DNA cleaving agent produced by this reagent, and the application of the Fenton reaction to the determination of the structure of the Holliday junction, the four-stranded DNA molecule that is a key intermediate in recombination. The cleavage pattern of the Holliday junction has pseudo-twofold symmetry, putting important constraints on possible structures. 相似文献
20.
对锰离子参与类Fenton反应机理的研究 总被引:1,自引:0,他引:1
锰主要以Mn2 形式存在,有人发现其具有与其他过渡性金属离子截然相反的抗氧化活性,采用自旋捕捉-ESR技术、芳环羟基化反应-高效液相色谱(HPLC)法和琼脂糖电泳法三种方法研究Mn2 参与类Fenton反应的情况时,均检测到Mn2 与H2O2反应产生.OH,Mn2 与H2O2反应可以发生类Fenton反应,产生.OH。这一现象的产生可能是Mn2 引起生物体内氧化损伤之故。同时显示,Mn2 的类Fenton反应是否产生.OH与反应过程Mn2 以及其他成分浓度有关(如高浓度抑制,低浓度促进),为诸多文献中Mn2 作为促氧化剂还是抗氧化剂的争论提供了可能解释。同时Mn2 能引起.OH持续低量的产生为一些慢性疾病的发生提供了合理的解释。 相似文献