The involvement of iron in lipid peroxidation. Importance of ferric to ferrous ratios in initiation

Intense lipid peroxidation of brain synaptosomes initiated with Fenton's reagent (H2O2 + Fe2+) began instantly upon addition of Fe2+ and preceded detectable OH. formation. Although mannitol or Tris partially blocked peroxidation, concentrations required were 10(3)-fold in excess of OH. actually formed, and inhibition by Tris was pH dependent. Lipid peroxidation also was initiated by either Fe2+ or Fe3+ alone, although significant lag phases (minutes) and slowed reaction rates were observed. Lag phases were dramatically reduced or nearly eliminated, and reaction rates were increased by a combination of Fe3+ and Fe2+. In this instance, lipid peroxidation initiated by optimal concentrations of H2O2 and Fe2+ could be mimicked or even surpassed by providing optimal ratios of Fe3+ to Fe2+. Peroxidation observed with Fe3+ alone was dependent upon trace amounts of contaminating Fe2+ in Fe3+ preparations. Optimal ratios of Fe3+:Fe2+ for the rapid initiation of lipid peroxidation were on order of 1:1 to 7:1. No OH. formation could be detected with this system. Although low concentrations of H2O2 or ascorbate increased lipid peroxidation by Fe2+ or Fe3+, respectively, high concentrations of H2O2 or ascorbate (in excess of iron) inhibited lipid peroxidation due to oxidative or reductive maintenance of iron exclusively in Fe2+ or Fe3+ form. Stimulation of lipid peroxidation by low concentrations of H2O2 or ascorbate was due to the oxidative or reductive creation of Fe3+:Fe2+ ratios. The data suggest that the absolute ratio of Fe3+ to Fe2+ was the primary determining factor for the initiation of lipid peroxidation reactions.     

邻苯二甲酸二丁酯对翡翠贻贝抗氧化酶及脂质过氧化水平的影响

实验室条件下,研究了不同浓度邻苯二甲酸二丁酯(DBP)长期胁迫(15 d)对翡翠贻贝内脏团和外套膜抗氧化酶(超氧化物歧化酶SOD、过氧化氢酶CAT)及脂质过氧化(LPO)水平(以MDA含量表示)的影响,以及受胁迫翡翠贻贝在清洁海水中恢复阶段上述生化指标的变化特征.结果表明:胁迫阶段,0.5和2.5 mg.L-1DBP下翡翠贻贝内脏团SOD活性表现为先抑制后逐渐恢复,12.5和62.5 mg.L-1下则持续受到显著抑制;不同浓度组CAT活性均明显被抑制.LPO水平明显升高.外套膜中,2.5 mg.L-1下SOD活性受到持续诱导,其他浓度组则先被抑制,后随曝露时间延长逐渐被诱导;各浓度组CAT的变化波动较大,没有明显规律;而LPO水平明显升高.净化恢复阶段,12.5和62.5 mg.L-1DBP胁迫下的内脏团SOD和CAT活性恢复较慢,其LPO水平随时间延长逐渐恢复至对照组水平;外套膜中SOD活性呈持续升高趋势,CAT活性和LPO水平则随时间延长恢复到对照组水平.     

模拟酸雨及其酸化土壤对小麦幼苗膜脂过氧化水平的影响

以小麦为试材,采用盆栽的方法研究了模拟酸雨及其酸化土壤对小麦幼苗膜脂过氧化水平的影响。结果表明以黄棕壤为材料通过模拟酸雨的淋溶,引起了土壤酸化和盐基流失。当模拟酸雨的pH值由5.6下降到2.5时,土壤pH值由6.06下降到3.41,土壤中交换性盐基总量便从56.5mmol/kg下降到41.1mmol/kg。将小麦幼苗栽培在该酸化土壤上,并分别用5种不同pH值的模拟酸雨喷淋地上器官,导致小麦幼苗体内的膜脂过氧化水平增高和保护酶的活性变化。其中模拟酸雨喷淋小麦幼苗对叶片中的膜脂过氧化水平及保护酶活性的影响大于酸化土壤对其产生的影响。而酸化土壤对小麦幼苗根系中的膜脂过氧化水平和保护酶活性的影响大于模拟酸雨喷淋对其产生的影响。同时,pH≤3.0的高强度酸雨以及由其产生的酸化土壤(T4、T5土壤)对小麦幼苗的膜脂过氧化水平和保护酶的活性产生了严重的影响。     

The effect of chronic alcohol feeding on lipid peroxidation in microsomes: lack of relationship to hydroxyl radical generation

Chronic alcohol feeding causes microsomal induction including increased generation of hydroxyl radicals. Ethanol induced liver injury may be mediated by lipid peroxidation for which hydroxyl radicals have been proposed as major mediators. Ethanol promotes lipid peroxidation when given acutely but also may serve as a hydroxyl radical scavenger. Therefore, we studied the acute and chronic effects of alcohol on microsomal lipid peroxidation and hydroxyl radical generation. Chronic alcohol feeding in rats increased microsomal generation of hydroxyl radicals but lipid peroxidation of endogenous lipid was inversely related to hydroxyl radical generation. Ethanol (50mM) had a slight inhibitory effect on hydroxyl radical production in peroxidizing microsomes, no effect on endogenous lipid peroxidation and enhanced the lysis of RBCs added as targets of peroxidation. Enhanced microsomal generation of hydroxyl radicals following chronic alcohol feeding is not an important mediator of lipid peroxidation.     

The requirement for iron (III) in the initiation of lipid peroxidation by iron (II) and hydrogen peroxide

The initiation of lipid peroxidation by Fe2+ and H2O2 (Fenton's reagent) is often proposed to be mediated by the highly reactive hydroxyl radical. Using Fe2+, H2O2, and phospholipid liposomes as a model system, we have found that lipid peroxidation, as assessed by malondialdehyde formation, is not initiated by the hydroxyl radical, but rather requires Fe3+ and Fe2+. EPR spin trapping with 5,5-dimethyl-1-pyrroline-N-oxide and the bleaching of para-nitrosodimethylaniline confirmed the generation of the hydroxyl radical in this system. Accordingly, catalase and the hydroxyl radical scavengers mannitol and benzoate efficiently inhibited the generation and the detection of hydroxyl radical. However, catalase, mannitol, and benzoate could either stimulate or inhibit lipid peroxidation. These unusual effects were found to be consistent with their ability to modulate the extent of Fe2+ oxidation by H2O2 and demonstrated that lipid peroxidation depends on the Fe3+:Fe2+ ratio, maximal initial rates occurring at 1:1. These studies suggest that the initiation of liposomal peroxidation by Fe2+ and H2O2 is mediated by an oxidant which requires both Fe3+ and Fe2+ and that the rate of the reaction is determined by the absolute Fe3+:Fe2+ ratio.     

Site-specific mechanisms of initiation by chelated iron and inhibition by alpha-tocopherol of lipid peroxide-dependent lipid peroxidation in charged micelles.

To obtain information on the role of iron-catalyzed lipid peroxidation in the presence of the small amount of lipid peroxide in deterioration of biological membranes, we examined factors affecting peroxidation of fatty acids in charged micelles. Peroxidation of linoleic acid (LA) was catalyzed by Fe2+ via reductive cleavage of linoleic acid hydroperoxide (LOOH) in negatively charged sodium dodecyl sulfate micelles, but not in positively charged tetradecyltrimethylammonium bromide (TTAB) micelles. However, this Fe2(+)-induced, LOOH-dependent lipid peroxidation could be induced in TTAB micelles in the presence of a negatively charged iron chelator, nitrilotriacetic acid (NTA). The linoleic acid alkoxy radical (LO.) generated by the LOOH-dependent Fenton reaction was also trapped by N-t-butyl-alpha-phenylnitrone at the surface of TTAB micelles in the presence of NTA, but not in its absence. The degradation rates of two spin probes, N-oxyl-4,4'-dimethyloxazolidine derivatives of stearic acid (5-NS and 16-NS), were investigated to determine the site of production of radicals formed during LOOH-dependent lipid peroxidation. The rate of consumption of 16-NS during the LOOH-dependent Fenton-like reaction was higher in TTAB micelles containing LA than in those containing lauric acid (LauA), although the rates of formation of LO. in the two types of fatty acid micelles were similar. The rates of 5-NS consumption in LA and LauA micelles were almost the same and were as low as that of 16-NS consumption in LauA micelles. 16-NS was more inhibitory than 5-NS of LOOH-dependent lipid peroxidation, and this inhibition was associated with its higher consumption of 16-NS than of 5-NS. alpha-Tocopherol inhibited NTA-Fe2(+)-induced LOOH-dependent lipid peroxidation in TTAB micelles, and was oxidized during this inhibition process. The rate and amount of alpha-tocopherol oxidized by the LOOH-dependent Fenton reaction were higher in LA micelles than in LauA micelles. alpha-Tocopherol inhibited the consumption of 16-NS during NTA-Fe2(+)-induced LOOH-dependent lipid peroxidation more effectively than that of 5-NS. The distribution of the chromanol moiety of alpha-tocopherol was studied by the fluorescence quenching method. There was no difference between Stern-Volmer plots of the quenchings of alpha-tocopherol fluorescence by 5-NS and 16-NS. From these results, we discuss the mechanism of induction of LOOH-dependent peroxidation of LA and the mechanism of the antioxidant effects of alpha-tocopherol on it from the viewpoint of site-specific reaction.     

Inhibition of lipid peroxidation by paraquat: site of inhibition in the cytochrome P-450-dependent steroid hydroxylase system from bovine adrenal cortex mitochondria

We have found that NADPH-dependent lipid peroxidation in bovine adrenal cortex mitochondria is strongly inhibited by paraquat. The site of the inhibition of the lipid peroxidation by paraquat has been examined. Paraquat neither inhibits NADPH-2,6-dichlorophenolindophenol nor NADPH-cytochrome c reductase activities. However, paraquat is able to retard the rate of reduction of cytochrome P-450 by NADPH. The spectrophotometric measurements provide the first evidence that lipid peroxidation in adrenal cortex mitochondria involves cytochrome P-450 and that the inhibitory effect of paraquat on lipid peroxidation is due to reoxidation of reduced cytochrome P-450 by the reagent.     

镉对小麦幼苗脂质过氧化和保护酶活性的影响

小麦幼苗经镉胁迫后,随着镉浓度的增高,叶征和根系中的膜脂过氧化产物丙二醛（ＭＤＡ）的含量和过氧物酶（ＰＯＤ）活性明显升高,超氧物歧化酶（ＳＯＤ）活性也有所提高。叶片中ＭＤＡ积累量和ＳＯＤ活性都高于根,而ＰＯＤ活性则是根高于叶片。随幼苗生长时间延长,叶片和根中的ＭＤＡ积累量增加,而ＳＯＤ活性却降低。     

Antioxidant status and lipid peroxidation in hemodialysis patients undergoing erythropoietin and erythropoietin-vitamin E combined therapy

In this study, plasma and red blood cell (RBC) antioxidant status and plasma lipid peroxidation were investigated in 46 hemodialysis patients. In addition, the effect of erythropoietin (EPO) and EPO-vitamin E combination therapy on plasma and RBC antioxidant status, and plasma lipid peroxidation were examined.

There were 10 healthy subjects in the control group and 10 hemodialysis patients in the untreated group. The third group included 36 hemodialysis patients that were given EPO (100 U/kg) for 3 months, 3 times per week. The fourth group included 36 hemodialysis-patients from the EPO group that were given EPO at a 50% decreased dose + vitamin E (300 mg/day) for 3 months.

MDA levels in the untreated group, the EPO group and the EPO + vitamin E groups were found to be higher than the control group (p<0.001, in both). Furthermore, MDA levels in both of the treatment groups were lower when compared to the untreated group (p<0.001, in both). Plasma vitamin E levels in the untreated, the EPO group and EPO + vitamin E groups were lower than the control group (p<0.001). In contrast, plasma vitamin E levels in the treatment groups were higher in comparison with the control group (p<0.05). SOD activities in the untreated, the EPO group and the EPO + vitamin E groups were found to be lower than the control group (p<0.001). SOD activities in the treatment groups were higher than the control group (p<0.001). The SOD activities in the EPO + vitamin E group increased when compared to the EPO group (p<0.001). CAT activities in the untreated, the EPO group and the EPO + vitamin E groups were found to be lower than the control group (p<0.001 in untreated and EPO groups, p<0.01 in EPO + vitamin E group). CAT activities in EPO and EPO + vitamin E groups were increased when compared to the untreated group (p<0.01).

In conclusion, our findings have shown that antioxidant status decreased and lipid peroxidation increased in hemodialysis patients. EPO has an antioxidant effect on the RBC and plasma antioxidant status, and plasma lipid peroxidation. These effects were moderately increased by the combination of vitamin E and EPO.     

大鼠多囊卵巢颗粒细胞凋亡时MDA含量及SOD活性的变化

目的和方法以大鼠多囊卵巢（PCO）为动物模型,观察了PCO大鼠卵泡颗粒细胞凋亡的发生率,并检测了颗粒细胞凋亡时丙二醛（MDA）的含量及超氧化物歧化酶（SOD）活性的变化。结果①PCO颗粒细胞凋亡发生率明显高于正常（P<0．001）;②PCO颗粒细胞发生凋亡时,细胞内MDA含量增加而SOD活性降低。结论大鼠PCO颗粒细胞的凋亡可能与细胞内MDA含量增加和SOD活性降低有关。     

Influence of acute heat stress on the development of GABAergic neurons in HPA-axies of mouse embryos

The damaging effects of acute heat stress in pregnant mice is well known, however, very little is known about the impact of heat on embryonic neurodevelopment and its dependence on the dam's physiological response to the stress. To study the changes in GABAergic neurons expression on the hypothalamo-pituitary-adrenocortical axis (HPA-axis), superoxide dismutase (SOD) activities and malondialdehyde (MDA) levels were measured in mouse embryos on E13-17 following acute, maternal heat stress. Blood samples and amniotic fluid from pregnant mice, and homogenates of whole embryos and embryo brains were collected for SOD and MDA analyses. SOD and MDA activities were measured in embryo and the sections by staining with anti-γ-aminobutyric acid-A- α1 receptor (GABAA receptor- α1), anti-γ-aminobutyric acid-B-1 receptor (GABAB receptor-R1) and anti-glutamate decarboxylase-65(GAD-65) antibodies. After the pregnant mice underwent acute heat stress on E13, the embryonic GABAergic neurons and GABA receptor expression were triggered immediately as a result of induction of the HPA-axis. This expression recovered to normal levels consistent with the control groups. However, the expression of the GABAA receptor in embryonic adrenal gland decreased continually. The SOD activity decreased in the embryonic brain and increased in the amniotic fluid after the heat stress, whereas the MDA levels increased in the maternal plasma only. Therefore, the GABAergic neurons in the developing HPA-axis of the embryos are susceptible to heat stress and the enhancement of SOD activities in the amniotic fluid might be a protective mechanism.     

The role of iron in the initiation of lipid peroxidation

Iron is required for the initiation of lipid peroxidation. Evidence is presented that lipid peroxidation requires both Fe3+ and Fe2+, perhaps with oxygen to form a Fe3+-dioxygen-Fe2+ complex. Other mechanisms of initiation, mostly involving the iron-catalyzed formation of hydroxyl radical, are described and discussed from both theoretical and experimental view points.     

Effect of drought on biomass, protein content, lipid peroxidation and antioxidant enzymes in two sesame cultivars

The effects of drought on growth, protein content, lipid peroxidation, superoxide dismutase (SOD), peroxidase (POX), catalase (CAT) and polyphenol oxidase (PPO) were studied in leaves and roots of Sesamum indicum L. cvs. Darab 14 and Yekta. Four weeks after sowing, plants were grown under soil moisture corresponding to 100, 75, 50 and 25 % field capacity for next four weeks. Fresh and dry masses, and total protein content in leaves and roots decreased obviously under drought. However, several new proteins appeared and content of some proteins was affected. Measurement of malondialdehyde content in leaves and roots showed that lipid peroxidation was lower in Yekta than in Darab 14. Severe stress increased SOD, POX, CAT and PPO activities in leaves and roots, especially in Yekta. According to the present study Yekta is more resistant to drought than Darab 14.     

Induction of lipid peroxidation by oxalate in experimental rat urolithiasis

The function of lipid peroxidation and the antiperoxidative enzymes of rat liver and kidney were studied in stone formation induced by intraperitoneal administration of sodium oxalate (7 mg/100 g body weight). The animals sacrificed 3 and 12 h after administration of sodium oxalate had higher level of malondialdehyde in liver and kidney than control animals. A significantly pronounced release of malondialdehyde was observed in treated liver and kidney homogenates when incubated with either ferrous sulphate or hydrogen peroxide compared to control liver and kidney. Superoxide dismutase activity was increased only in liver and not in kidney in treated animals compared to the control. A highly significant decrease in catalase activity was observed in both liver and kidney of treated animals.     

Influence of salt stress on growth,lipid peroxidation and antioxidative enzyme activity in borage (Borago officinalis L.)

Abstract

The effects of increasing salt concentrations on the growth, electrolyte leakage, lipid peroxidation, and major antioxidant enzyme activities (superoxide dismutase, catalase, ascorbate peroxidase and glutathione reductase) of borage plants were investigated. Plants were grown in half strength of Hoagland nutrient solution added with 0, 25, 50, and 75 mM of NaCl. Most measured parameters were affected by salinity. Increasing salt levels caused a significant reduction in leaf area, stem length, stem diameter, flower number, and dry masses of different organs. Growth of borage plants, in terms of dry weight, was affected. As a consequence of salinity stress, lipid peroxidation and membrane permeability was increased. Antioxidant activity showed an increase in the activity of superoxide dismutase, a non-induced activity of catalase and ascorbate peroxidase, and a slight increase in glutathione reductase activity. The results indicate that borage plants appear to be sensitive to salt stress, since enzymes related to antioxidant enzymatic defense system in treated leaves should be highly active.     

Chromosome aberration and lipid peroxidation in chromium-exposed workers

Chromosome aberration frequency and lipid peroxidation levels were analysed to investigate their efficacy as biological markers for monitoring the genotoxicity and oxidative damage in Korean chromium (Cr)-exposed workers. Fifty-one Cr-exposed workers and 31 age-matched controls in ten chrome-plating plants were sampled. The Cr level was measured in the workers' blood and urine, and in the ambient air at the workplaces. The conventional Giemsa staining method and fluorescence in situ hybridization (FISH) technique were used for chromosome aberration analysis. Spectrum green whole chromosome paint specific for chromosome 4 was used in the FISH procedure. As for lipid peroxidation, malondialdehyde (MDA) was measured in the blood plasma as thiobarbituric acid-reactive substances (TBARS). The blood Cr concentration was statistically correlated with both the frequency of chromatid exchange and the total frequency of chromosome/chromatid breaks and exchanges, as detected by the Giemsa staining. Meanwhile, the frequency of translocation, as detected by the FISH technique, was significantly higher in the Cr-exposed workers than in the controls and it correlated with the blood Cr concentration. Although the concentration of MDA, the metabolite of lipid peroxidation, in the exposed workers was higher than that in the controls, no statistically significant correlation between the MDA level and the blood or urine Cr levels was observed. Accordingly, the genotoxicity and oxidative damage (plasma lipid peroxidation) in the Korean Cr-exposed workers were consequential at quite low exposure levels, plus chromosome rearrangement, especially translocation, was clearly evident as a biological response marker for Cr exposure based on a significant positive correlation between the translocations detected by FISH and the Cr in the blood.     

双丹胶囊对大鼠脑缺血再灌注损伤保护作用研究

目的：观察双丹胶囊对大鼠局灶性脑缺血再灌注损伤的脑梗死体积、自由基变化的影响,探讨双丹胶囊对脑缺血损伤的保护作用。方法：复制大鼠中动脉缺血再灌注模型,分别给药干预,在给药后观察行为学、脑梗死率、脑指数、脑含水量、SOD、MAD等指标。结果：双丹胶囊可改善动物的神经行为学评分,明显降低动物的脑梗死率、脑指数、脑含水量、提高脑组织SOD活性、降低MDA含量,并成剂量依赖。结论：双丹胶囊对脑缺血再灌注损伤具有保护作用。     

A chromium-tolerant plant growing in Cr-contaminated land

In this paper, a kind of Typhaceae plant species, Typha angustifolia L, with a high tolerance to Cr is described. Experiments were carried out to examine its ability to tolerant Cr and its physiological response. The results showed that there was no difference in growth, plant height, and biomass response to external Cr (VI) between the plants exposed to 100 microM Cr (VI) and control (0 microM), while increasing Cr levels to 200-800 microM induced a significant decrease in plant height and biomass, but no significant injury was detected, even for the plants exposed to 800 microM Cr. Chromium induced significant increases in superoxide dismutase (SOD) and peroxidase (POD) activities. Meanwhile, a significantly positive correlation was found between Cr and Mn or Cu in leaves and roots, respectively. The Cr tolerance of the plant appeared to be associated with the enhancement of SOD and POD activities and the improvement in uptake and translocation of the essential microelements.     

Fasciola hepatica: effects on the antioxidative properties and lipid peroxidation of rat serum

Fasciola hepatica infection is accompanied by increased formation of reactive oxygen species. The aim of this study was to analyze antioxidative properties of rat serum in the course of fasciolosis. Wistar rats were infected per os with 30 metacercariae of F. hepatica. Activities of antioxidant enzymes and concentrations of non-enzymatic antioxidants in serum were determined at 4, 7, and 10 weeks post-infection (wpi). Activity of superoxide dismutase (Cu, Zn-SOD) significantly decreased (by 35% during the migratory phase, by 40 and 23% at 7 and 10 wpi, respectively), while glutathione reductase activity significantly increased (by 62, 65, and 41%, at 4, 7, and 10 wpi, respectively). No significant changes were found in the activity of glutathione peroxidase. Significant decreases in concentrations of reduced glutathione, vitamins C, E, and A were observed, particularly during the migratory phase of fasciolosis (at 4 wpi). These changes were accompanied by enhancement of lipid peroxidation processes as evidenced by increased levels of malondialdehyde (MDA) and 4-hydroxynonenal (4-HNE). Concentrations of MDA and 4-HNE at 4 wpi increased by 38% and by 59%. MDA increased by 51% at 7 wpi and by 79% at 10 wpi, while 4-HNE increased by 87 and 118%, respectively. The results indicate that fasciolosis is associated with enhanced oxidative reactions and reduced antioxidant defense capability of rat serum.