The Influence of Superoxide Dismutase and Glutathione Peroxidase Deficiencies on Noise-Induced Hearing Loss in Mice

One consequence of noise exposure is increased production of reactive oxygen species (ROS), such as superoxide, hydrogen peroxide, and hydroxyl radicals, in the cochlea. ROS can cause oxidative damage to diverse cellular components, including membranes, proteins, and DNA, if they are not "neutralised" by antioxidant defences. Two important enzymes of the cochlear antioxidant defense system are cytosolic copper/zinc superoxide dismutase (SOD1) and selenium-dependent glutathione peroxidase (GPx1). These metalloenzymes work together to regulate ROS production in virtually every cell in the body, and they may be important for limiting cochlear damage associated with aging and acoustic overexposure. In this chapter, we describe a series of experiments using mice with targeted deletions of Sod1 or Gpx1, the mouse genes that code for SOD1 and GPx1, respectively, to study the cellular mechanisms underlying noise-induced hearing loss (NIHL). The results from Sod1 and Gpx1 knockout mice provide insights into the link between endogenous levels of antioxidant enzymes and susceptibility to NIHL.     

In vitro antioxidant and free radical scavenging activity of Cyperus rotundus

Cyperus rotundus (Family Cyperaceae) is used both as a functional food and as a drug. In this study, the antioxidative potential of a hydroalcoholic extract of C. rotundus (CRE) was evaluated by various antioxidant assays, including antioxidant capacity by the phosphomolybdenum method, total antioxidant activity in linoleic acid emulsion systems, 1,1-diphenyl-2-picrylhydrazyl (DPPH), superoxide, hydroxyl radicals, and nitric oxide (NO) scavenging. We further evaluated the reducing potential of the extract as well as Fe(2+)/ascorbate-induced lipid peroxidation in rat liver homogenate. These various antioxidant activities were compared to standard antioxidants such as butylated hydroxytoluene, tocopherol, L-ascorbic acid, and catechin. Total phenolic and flavonoid content of CRE was also determined by a colorimetric method. The extract exhibited high reduction capability and powerful free radical scavenging, especially against DPPH and superoxide anions as well as a moderate effect on NO. CRE also showed inhibited lipid peroxidation in rat liver homogenate induced by Fe(2+)/ascorbate and prevented deoxyribose degradation in both non-site-specific and site-specific assays showing the hydroxyl radical scavenging and metal chelating activity of the hydroalcoholic extract. Moreover, the peroxidation inhibiting activity of CRE was demonstrated in the linoleic acid emulsion system. These results clearly established the antioxidative potency of C. rotundus, which may account for some of the medical claims attributed to this plant.     

Free radicals: important cause of pathologies refer to ageing

Free radical are highly reactive chemical species with an unpaired electron in an atomic or molecular orbital. In biological systems, the most important free radicals are superoxide anion and hydrogen peroxide; in the presence of transition metals such as iron, copper and manganese both these free radicals produce hydroxyl radicals. Free radicals attack proteins, nuclei acids and membranes containing large quantities of polyunsaturated fatty acids. Because of their toxicity, the organism has developed ways to deactivate them. The superoxide dismutase enzyme (SOD) catalyzes dismutation of the superoxide radical into hydrogen peroxide and oxygen hydrogen peroxide is in turn reduced to water and oxygen by peroxidase glutathione and catalase enzymes. The production of radicals in the brain is due to catecholamine metabolism such as dopamine and norepinephrine and is increased by the presence of transition metals and by a deficiency of antioxidant agents such as vitamin E. Two main groups of dementia exist in older age: the multi-infarctual dementias, caused by cerebrovascular disorders and the primary degenerative disorders such as Alzheimer, where no vascular disease is evident. Free radicals play an important role in Parkinson's disease, in Alzheimer's disease and in stroke. The value of SOD and CAT activity following the above mentioned degenerative diseases differ among the various studies carried out. In Alzheimer's disease, the value of SOD activity probably increases in the neuropathologically involved areas. In stroke, the SOD value does not vary either in the ischemic area or in the peri-infarctual one during the first 24 hrs after lesion, while the CAT value decreases.     

Antioxidative effects of glycoprotein isolated from Solanum nigrum L

Glycoprotein from Solanum nigrum L. (SNL glycoprotein) was isolated and tested for antioxidative effects on oxygen free radicals using a 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay. The free radical scavenging activities of the SNL glycoprotein are optimal in acidic pH and up to 60 degrees C. However, it has minimal activities in the presence of EDTA, although such activities are not dependent on M(2+) ions (Ca(2+), Mn(2+), and Mg(2+)) in the presence of EDTA. Interestingly, when SNL glycoprotein was treated with deactivation agents (pronase E and NaIO(4)), the DPPH radical scavenging activity was decreased compared with the SNL glycoprotein treatment alone. The antioxidative effects of SNL glycoprotein on superoxide anion and hydroxyl radical under optimal conditions revealed that SNL glycoprotein has remarkable scavenging effects on both radicals, but exhibited slightly higher scavenging effects on superoxide anion generated by the enzymatic hypoxanthine/xanthine oxidase system than on hydroxyl radicals generated by the Fenton reaction. However, SNL glycoprotein was more effective against hydroxyl radials in cell cultures (NIH/3T3). Consequently, 20 microg/mL SNL glycoprotein has a scavenging ability against superoxide anion corresponding to that of ascorbic acid. On the other hand, its hydroxyl radical scavenging activity corresponds to 0.1 microg/mL catalase. From these results, we suggest that SNL glycoprotein has potent antioxidative potential.     

Evaluation of antioxidant properties of a new compound, pyrogallol-phloroglucinol-6,6'-bieckol isolated from brown algae, Ecklonia cava

In this study, antioxidant and free radical scavenging activities of the natural antioxidative compound, pyrogallol-phloroglucinol-6,6''-bieckol (PPB) isolated from brown algae, Ecklonia cava was assessed in vitro by measuring the radical scavenging activities (DPPH, alkyl, hydroxyl, and superoxide) using electron spin resonance (ESR) spectrometry, intracellular reactive oxygen species (ROS) scavenging activity, and DNA damage assay. According to the results of these experiments, the scavenging activity PPB against difference radicals was in the following order: DPPH, alkyl, hydroxyl, and superoxide radicals (IC₅₀; 0.90, 2.54, 62.93 and 109.05 µM). The antioxidant activities of PPB were higher than that of the commercial antioxidant, ascorbic acid. Furthermore, PPB effectively inhibited DNA damage induced by H₂O₂. These results suggest that the natural antioxidative compound, PPB, can be used by the natural food industry.     

芒果叶总黄酮含量及抗氧化作用测定

目的研究芒果叶总黄酮(MLF)的体外抗氧化作用。方法采用超声辅助乙醇浸提方法提取芒果叶中的黄酮类化合物,评价芒果叶总黄酮的体外清除自由基能力及抗脂质过氧化活性。结果 芒果叶提取物中总黄酮的含量为38.35%,对羟自由基和超氧阴离子自由基具有一定的清除能力;维生素C和芒果叶总黄酮对羟自由基的半数清除浓度(IC50)分别为0.546 9、2.973 9 mg/mL,对超氧阴离子自由基清除的IC50分别为0.253 0、3.861 2 mg/mL,清除能力与浓度有明显的量效关系;芒果叶总黄酮浓度(x)与抑制率(y)之间拟合曲线,得出回归方程为:y=5.263 9 ln(x) + 50.947,R²=0.982 1,表明芒果叶总黄酮在1~10 mg／mL剂量范围内能有效抑制肝组织自发性脂质过氧化。结论 芒果叶总黄酮具有清除自由基及抗脂质过氧化和红细胞溶血作用。     

Effects of active oxygen species on damage to and prostaglandin synthesis in cultured rat gastric cells.

Active oxygen species cause gastric mucosal damage in vivo. However, it is not known if these species are directly cytotoxic toward gastric cells. Prostaglandins have important physiological roles in the gastric mucosa, including direct cell protection against damaging factors. So, to find if active oxygen species affect prostaglandin synthesis in gastric mucosal cells is important, but this also is not known. This study was done to investigate the effects of such species on damage to and prostaglandin synthesis in cultured mucus-producing cells from rat gastric mucosa. Active oxygen species were produced by the addition of xanthine and xanthine oxidase to the culture medium. Cytotoxicity was assayed by 51Cr release. Xanthine (1 mM) and xanthine oxidase (100 mU/ml) increased specific 51Cr release as the thiobarbituric acid reactants increased. This increase in 51Cr release was inhibited by catalase, a scavenger of hydrogen peroxide, or dimethyl sulfoxide, a scavenger of hydroxyl radicals, but not by superoxide dismutase, a scavenger of superoxide, nor deferoxamine, an inhibitor of hydroxyl radical generation. Catalase, dimethyl sulfoxide, and superoxide dismutase each had no effect on prostaglandin E2 synthesis when xanthine and xanthine oxidase were not added. In the presence of xanthine and xanthine oxidase, catalase and dimethyl sulfoxide stimulated the synthesis of prostaglandin E2 and superoxide dismutase inhibited it. Indomethacin, a prostaglandin synthetase inhibitor, did not affect the decrease in 51Cr release caused by catalase in the presence of xanthine and xanthine oxidase, but it abolished the decrease caused by dimethyl sulfoxide. These results suggest that hydrogen peroxide, but not superoxide nor hydroxyl radicals, is involved in damage to cultured rat gastric cells, and that superoxide stimulates prostaglandin E2 synthesis, but that hydrogen peroxide inhibits it. Protection of the cells by dimethyl sulfoxide may be related to stimulation of prostaglandin E2 synthesis in the cells, but not via scavenging hydroxyl radicals.     

Free radical reactions and the inhibitory and lethal actions of high-pressure gases

This study was designed to test whether free radicals are involved in the deleterious effects of compressed gases on cells. The actions of xenon, nitrous oxide, argon, nitrogen, helium, and oxygen and their effects on the toxicity of paraquat (methyl viologen) were studied using Escherichia coli. Growth of E. coli in trypticase-soy broth in an atmosphere of 1.36 MPa (13.6 atm) N2O resulted in an induction of superoxide dismutase (SOD). In addition, when SOD was induced by oxygen, the resulting cells had increased resistance to the killing action of N2O. The toxicity of paraquat was increased in the presence of N2O but not He, N2, or Ar. However, addition of any of the latter three gases to N2O resulted in increased toxicity of paraquat beyond that due to N2O alone. Oxygen is known to increase the reaction of paraquat radicals within cells and to reduce leakage of the radicals out through the cell membrane. N2O and Xe seem to have this same action, and He, N2, or Ar could enhance the actions of N2O, Xe, or O2. The data indicate that the inhibitory and lethal actions of these gases may be due to enhanced reactivity of radicals with cell components and reduced leakage of the radicals to the environment.     

Antioxidant activity of methanolic extract of emblica fruit (Phyllanthus emblica L.) from six regions in China

The phenolic contents of methanolic extracts of emblica (Phyllanthus emblica L.) fruit from six regions in China were measured in this work. The antioxidant activities of these extracts were also evaluated. Total phenolic content was ranged from 81.5 to 120.9 mg gallic acid equivalents (GAE)/g and the flavonoid content was varied from 20.3 to 38.7 mg quecetin equivalents (QE)/g, while proanthocyanidin content was ranged from 3.7 to 18.7 mg catechin equivalents (CE)/g. Among all the methanolic extracts analyzed, the Huizhou sample exhibited a significantly higher phenolic content than other samples (P<0.05). The antioxidant activities were evaluated by in vitro experiments using scavenging assays of 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals, hydroxyl radicals, and superoxide anion radicals, chelating ability of ferrous ion, reducing power, and inhibition capability of Fe (II)-induced lipid peroxidation, respectively. The Huizhou sample was found to have the strongest antioxidant activities in scavenging DPPH radicals, superoxide anion radicals, and had the highest reducing power, while the Chuxiong sample showed the best performance in chelating iron and inhibiting lipid peroxidation. Furthermore, the Chuxiong sample exhibited a stronger inhibition activity of the hydroxyl radicals compared with other samples. The high correlation coefficient was existed between the phenolic content and superoxide anion radical scavenging activity, but no significant correlation was found between the former and hydroxyl radical scavenging activity. Methanolic extracts of emblica fruit from some selected regions exhibited stronger antioxidant activities compared to those of the commercial compounds (quercetin and BHA). It might be considered as a potential plant source of antioxidants.     

乳杆菌抑制黄曲霉孢子萌发的研究

为探讨乳杆菌抑制黄曲霉孢子萌发的可能机制 ,在乳杆菌培养液、乳杆菌培养液的上清液和乳杆菌的细胞悬浮液中接种一定量的黄曲霉孢子 ,每隔 4h分别取少量液体在PDA平板上进行霉菌计数 ,直到 30h。 2 8℃培养 72h后计数萌发的霉菌孢子。结果表明 :只有乳杆菌培养液组对黄曲霉孢子萌发有显著的抑制作用 ,说明乳杆菌抑制黄曲霉孢子萌发的可能机制是低pH值 ,乳杆菌的代谢产物与微生物间竞争多因素协同作用的结果。     

Production and consumption of reactive oxygen species by fullerenes

Reactive oxygen species (ROS) are one of the most important intermediates in chemical, photochemical, and biological processes. To understand the environmental exposure and toxicity of fullerenes better, the production and consumption of ROS (singlet oxygen, superoxide, hydrogen peroxide, and hydroxyl radicals) by Buckminster fullerene (C(60) ) and fullerenol were investigated in aqueous systems. Fullerenol exhibits higher photoproduction efficiency of singlet oxygen and superoxide than aqueous suspensions of C(60) aggregates (aqu/nC(60) ), and this higher efficiency results in higher steady-state concentrations of these two ROS. Transmission electron microscopy indicates that the C(60) molecules in aqu/nC(60) are much more closely packed than the C(60) cages in fullerenol. These observations provide additional evidence that the lower ROS production efficiency of aqu/nC(60) is attributable primarily to efficient self-quenching of C(60) triplet states. Production of singlet oxygen by aqu/nC(60) is accelerated by increasing oxygen concentration and in part is sensitized by fluorescent photoproducts that accumulate during irradiation. The fullerenes react slowly with singlet oxygen (second-order rate constant <4?×?10(5) M(-1) s(-1) ), but react rapidly with hydroxyl radicals (second-order rate constants of 5.4?×?10(9) and 4?×?10(8) M(-1) s(-1) for aqu/nC(60) and fullerenol, respectively). These results show that environmental conditions, including light exposure and oxygen concentration, have the potential to impact the generation of toxic ROS by fullerenes.     

Antioxidative role of selenium on some tissues of (Cd2+), Cr3+)-induced rainbow trout

Chemical toxic pollutants (especially heavy metals) are important sources of reactive oxygen species (ROS) in biological systems. Membrane phospholipids of aerobic organisms are continually subjected to oxidant challenges from endogenous and exogenous sources, while peroxidized membranes and lipid peroxidation products represent constant threats to aerobic cells. The primary antioxidant protection against free radical and ROS is provided by the enzymes glutathione peroxidase (GSH-Px), superoxide dismutase (SOD) and catalase (CAT), respectively. The trace element selenium has been implicated in chemo-prevention and drug-resistance through reduction of oxidative stress. Selenium could prevent damage to the unsaturated fatty acid of subcellular membranes by lipid peroxidation induced by free radicals. The results reported here show that sodium selenite has an important contribution to antioxidative defense for the spleen and heart of rainbow trout. The ability of sodium selenite to prevent the oxidative stress induced by heavy metals (Cd(2+), Cr(3+)) in fish was rationalized.     

车前子对高脂血症大鼠脂质过氧化的影响

目的 :　探讨车前子对高脂血症大鼠脂质过氧化的影响。方法 :　采用大鼠血清及多脏器组织 ,观察车前子对高脂血症大鼠脂质过氧化的影响。结果 :　高脂对照组血清和心肌超氧化物歧化酶 ( SOD)活性显著降低 ,而脂质过氧化物 ( LPO)含量明显升高 ,血清和肝脏的过氧化氢酶( CAT)、谷胱苷肽过氧化物酶 ( GSH-Px)活性明显降低。补充车前子后显著升高血清及心肌组织SOD活性 2 2 %和 2 8% ,以及肝脏 GSH-Px活性 2 2 %。而使血清及心肌的 LPO含量分别下降 3 2 %和 1 7%。结论 :　 1 5 g/kg车前子可提高高脂血症大鼠体内抗氧化能力并免受自由基损伤     

Antioxidant effect of beta-carotene on lipid peroxidation and synergism with tocopherol in an emulsified linoleic acid model system

The antioxidant effect of beta-carotene was investigated in an emulsified linoleic acid model system with or without exposure to light. The effect of tocopherol on the antioxidant action of beta-carotene was also investigated. The antioxidant effect of beta-carotene was significant, and synergistically increased by the addition of rac alpha-tocopherol, in the dark but not in the light. It was#10; confirmed that superoxide anions (O(2)(-)) and hydrogen peroxide (H(2)O(2)) were formed during the light exposure, and that their formation could be significantly inhibited by the addition of tryptophan or mannitol, scavenger of hydroxyl radicals. These results strongly suggest direct participation of hydroxyl radicals, formed via the Haber-Weiss reaction, in the emulsified linoleic acid model system, with beta-carotene inhibition in the dark but an increase in the light. Regarding the mechanism of the antioxidant effect of beta-carotene, when beta-carotene is oxidized, the hydroperoxide in the acts as an electron acceptor and the hydroperoxide itself then decomposes.     

嗜酸乳杆菌与白色假丝酵母菌交互作用

目的对嗜酸乳杆菌与白色假丝酵母菌间的交互作用及pH、乳酸、乙酸、H2O2等对白色假丝酵母菌生长影响进行评价,为细菌性阴道病治疗后易转为外阴阴道假丝酵母菌病的原因提供依据。方法不同浓度嗜酸乳杆菌、白色假丝酵母菌混合培养;白色假丝酵母菌经不同pH值培养基培养、1%乳酸、1%乙酸、3%H2O2作用,检测其浓度变化,形态学观察白色假丝酵母菌形态变化。结果嗜酸乳杆菌浓度依赖性抑制白色假丝酵母菌生长,白色假丝酵母菌可促进嗜酸乳杆菌生长;pH值为3.5~5.5时,嗜酸乳杆菌具有较强的抑菌能力,当pH值为4.0时,嗜酸乳杆菌抑菌能力最强;乳酸、乙酸均对白色假丝酵母菌生长有明显抑制作用且具有浓度和时间依赖性,H2O2对白色假丝酵母菌生长并无直接抑制作用。结论嗜酸乳杆菌主要通过其代谢产物乳酸而不是H2O2直接产生抑菌作用,细菌性阴道病治疗后转为外阴阴道假丝酵母菌病的发生,主要与乳杆菌因某些因素生长受抑,导致数量降低及其产酸能力下降有关。     

Antioxidant and pro-oxidant effects of green tea extracts in oxygen radical absorbance capacity assay

Green tea extracts (GTEs) [water (GTE-W) and 75% ethanol (GTE-E)] were investigated to characterize their propensities to act as antioxidants or as pro-oxidants by analyzing oxygen radical absorbance capacity (ORAC) and scavenging capacity for hydroxyl radical. When 2,2'-azobis(2-amidinopropane) dihydrochloride was used for the generation of peroxyl radicals, both GTE-W and GTE-E exhibited strong concentration-dependent scavenging activity through donating protons, which could be explained by their reducing property. When hydroxyl radicals were generated through the addition of Cu(2+) and H(2)O(2), GTE-W and GTE-E exhibited antioxidant activity or pro-oxidant activity, depending on their concentrations, which might be attributed to the metal chelating activity, the scavenging activity on hydroxyl radical, and/or the pro-oxidant activity to generate some reactive oxygen species. When Cu(2+) without H(2)O(2) was used as an oxidant in the assay, the copper-initiated pro-oxidant activities of GTE-W and GTE-W was consistent with the availability of (-)-epigallocatechin and (-)-epicatechin to generate hydrogen peroxide and/or hydroxyl radical. The pro-oxidant activity of GTE-W and GTE-E was demonstrated by the deoxyribose assay. These results indicate that both GTE-W and GTE-E can have pro-oxidant activity at lower concentrations and antioxidant activity at higher concentrations in the ORAC and deoxyribose assays using generated hydroxyl radicals.     

Scavenging effects of methanolic extracts of broad beans on free-radical species

This report describes the antioxidant characteristics of methanolic extracts from broad beans (Vicia fava). The methanolic extracts of broad beans (MEBB) exhibited a marked scavenging effect on superoxide. MEBB also exerted scavenging activities on hydrogen peroxide and 1, 1-diphenyl-2-picrylhydrazyl radical. The radical scavenging activity of MEBB was highest when the scavenging effect of MEBB on Superoxide (IC₅₀ = 0.15 mg/ml) was examined. These results suggest that MEBB have effective activities both as a radical scavenger and as a hydrogen donor. The chelating activity of MEBB (0.70 mg/ml) on Fe²⁺ and Cu²⁺ was 31.2% and 28.5%, respectively. The antioxidant effect of MEBB on lipid peroxidation might be attributed to their properties of scavenging free-radical species and their chelating activity on metal ions. The antioxidant activity of MEBB against tert-butyl hydroperoxide (BHP)-induced oxidative stress in WI-38 cells was assessed. The activities of antioxidant enzymes such as superoxide dismutase (SOD), catalase, and glutathione peroxidase (GSH-Px) were measured as indices of oxidative stress. WI-38 cells incubated with 0.1 mM BHP for 2 hr exhibited the increase of SOD, catalase and GSH-Px activities over the control. When the cells incubated in MEBB (45–450 μg/ml) for 18 hr were subjected to a BHP challenge test, SOD activity returned to its control value or lower at all levels tested. When catalase activity was determined, a similar trend occurred except in the cells incubated in 112.5 μ g/ml MEBB. These results imply that MEBB inhibit oxidative stress in WI-38 cells.     

Antioxidative activity of hen egg ovalbumin hydrolysates

To evaluate the antioxidative activity of the hydrolysates of ovalbumin, the antioxidative activities of the enzymatic extracts were evaluated using three different methodologies scavenging assays such as superoxide anion, hydroxyl radical, and inhibitory oxidation of linoleic acid in vitro, and the activities of SOD, GSH-PX, CAT and the level of MDA were determined in serum and liver of aged mice induced by G-gal. The results showed that the hydrolysates had a distinctly inhibitory action to superoxide anion made by alkaline pyrogallic acid, HO. produced by Fenton reaction, the oxidation of linoleic acid in linoleic acid autoxidation system, and presented a positive correlation. The inhibition capacity of hydrolysates against superoxide anion and HO. were more than 45% and 56% respectively at the concentration 5 mg/mL. And the hydrolysates could significantly (p< 0.01) prevented the activities of SOD, GSH-Px, and CAT against reducing and all three concentrations could significantly (p< 0.01) decrease the MDA contents in the serum and liver of aged mice induced by G-gal. The antioxidative activity of high concentration was similar to that of control group.     

番石榴叶提取物对小鼠肝脏抗氧化作用研究

目的研究番石榴叶提取物抗氧化活性。方法干燥的番石榴叶分别用蒸馏水、65%和95%乙醇浸提,浸提液过滤、浓缩、干燥得到3种提取物。测定各提取物对羟自由基清除率和脂质过氧化抑制率。紫外分光光度法分析提取物中总黄酮含量,高效液相色谱(HPLC)和紫外可见吸收光谱法(UV)对黄酮类化合物初步鉴定。结果水、65%和95%乙醇提取物均具有清除羟自由基和抑制脂质过氧化作用,且呈剂量-效应关系,对羟自由基清除作用的半数有效浓度(EC50)分别为0.63、0.47和0.58g/L,对脂质过氧化抑制作用的半数有效浓度(EC50)分别为0.20、0.035和0.18g/L,总黄酮含量分别为3.28、30.71和55.98g/kg。结论番石榴叶水和乙醇提取物具有较强的抗氧化作用,其中黄酮类化合物可能是其功效成分之一。     

Effect of heavy metal stress on proline, malondialdehyde, and superoxide dismutase activity in the cyanobacterium Spirulina platensis-S5

This study investigated toxic impacts of lead, copper, and zinc over a concentration gradient of 0.05-0.20 mg/L on proline, malondialdehyde (MDA), and superoxide dismutase (SOD) in the cyanobacterium Spirulina platensis-S5. Despite a reduction in growth of the test microorganism, its MDA, SOD, and proline contents increased under the heavy metal stress, corresponding to the concentration of the metal ion in the culture medium. Increased amount of MDA was indicative of formation of free radicals in the test microorganism under heavy metals stress(es), while increased levels of SOD and proline pointed to the occurrence of a scavenging mechanism.