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1.
Lipid peroxidation initiated by superoxide-dependent hydroxyl radicals using complexed iron and hydrogen peroxide 总被引:3,自引:0,他引:3
J M Gutteridge 《FEBS letters》1984,172(2):245-249
Iron salts stimulate lipid peroxidation by decomposing lipid peroxides to produce alkoxyl and peroxyl radicals which initiate further oxidation. In aqueous solution ferrous salts produce OH. radicals, a reactive species able to abstract hydrogen atoms from unsaturated fatty acids, and so can initiate lipid peroxidation. When iron salts are added to lipids, containing variable amounts of lipid peroxide, the former reaction is favoured and OH. radicals contribute little to the observed rate of peroxidation. When iron is complexed with EDTA, however, lipid peroxide decomposition is prevented, but the complex reacts with hydrogen peroxide to form OH. radicals which are seen to initiate lipid peroxidation. Superoxide radicals appear to play an important part in reducing the iron complex. 相似文献
2.
1. Thiol oxidation by a lipid peroxide or hydrogen peroxide was as efficient in denatured non-haem proteins as in small thiols. Both peroxides were relatively ineffective in oxidizing haemoprotein thiols, especially at low pH. Increased amounts of haematin decreased greatly the efficiency of GSH oxidation by peroxides especially at low pH. 2. Other than the haematin ring, the thiol group was found to be probably the group in proteins most sensitive to modification by peroxides. 3. At low concentrations, the fatty acid moiety of a lipid peroxide appeared to impede thiol oxidation in proteins, probably by hydrophobic bonding to the protein, rather than to stimulate thiol oxidation by denaturing the protein and thereby increasing the exposure and reactivity of the thiol group. 4. The relative rates of thiol oxidation by peroxides in the different thiols were: haemoprotein thiols>small thiols>other protein thiols. In all cases, thiol oxidation was much more rapid by the lipid peroxide than by hydrogen peroxide. 相似文献
3.
J M Gutteridge 《Chemico-biological interactions》1985,56(1):113-120
The copper-containing protein caeruloplasmin is an important biological extracellular protein. By catalysing the oxidation of ferrous ions to the ferric state (ferroxidase activity) it can inhibit lipid peroxidation and the Fenton reaction. This activity is readily destroyed by heat-denaturation. When a ferric-EDTA complex is added to hydrogen peroxide, OH X radicals are formed in a reaction inhibitable by superoxide dismutase (SOD). This reaction is also inhibited by caeruloplasmin both before and after heat-denaturation, suggesting a non-catalytic scavenging role for the protein. A combination of ferroxidase and radical scavenging activities in fluids containing iron complexes and hydrogen peroxide, but no SOD or catalase, would make caeruloplasmin an important extracellular antioxidant. 相似文献
4.
T Okuda Y Matsuda M Sugawara S Sagisaka 《Bioscience, biotechnology, and biochemistry》1992,56(12):1911-1915
We treated leaves of winter wheat (Triticum aestivum L.) with cold, paraquat, or 3-amino-1,2,4-triazole and compared the responses. We assayed the activities of glucose-6-phosphate dehydrogenase, catalase, dehydroascorbate reductase and ascorbate free radical reductase and levels of hydrogen peroxide, glucose-6-phosphate, fructose-6-phosphate, ascorbate, dehydroascorbate, reduced and oxidized glutathione. With any of the three treatments, contents of cellular peroxides and hexose phosphates were raised. The content of ascorbate was lowered markedly by paraquat treatment, which produces active oxygen species, whereas such a decrease did not occur in other two treatments. When the plants were treated with 3-amino-1,2,4-triazole, which is a specific inhibitor of catalase, the content of oxidized glutathione increased severalfold. The glucose-6-phosphate dehydrogenase activity increased with all three treatments, but it decreased after glyphosate treatment, which does not stimulate the formation of peroxides. The activities of catalase and dehydroascorbate reductase were increased by the treatment of cold and paraquat, while 3-amino-1,2,4-triazole did not affect the dehydroascorbate reductase activity. The activity of ascorbate free radical reductase increased after treatment by paraquat only. 相似文献
5.
《Bioscience, biotechnology, and biochemistry》2013,77(12):1911-1915
We treated leaves of winter wheat (Triticum aestivum L.) with cold, paraquat, or 3-amino-1,2,4-triazole and compared the responses. We assayed the activities of glucose-6-phosphate dehydrogenase, catalase, dehydroascorbate reductase and ascorbate free radical reductase and levels of hydrogen peroxide, glucose-6-phosphate, fructose-6-phosphate, ascorbate, dehydroascorbate, reduced and oxidized glutathione. With any of the three treatments, contents of cellular peroxides and hexose phosphates were raised. The content of ascorbate was lowered markedly by paraquat treatment, which produces active oxygen species, whereas such a decrease did not occur in other two treatments. When the plants were treated with 3-amino-1,2,4-triazole, which is a specific inhibitor of catalase, the content of oxidized glutathione increased severalfold. The glucose-6-phosphate dehydrogenase activity increased with all three treatments, but it decreased after glyphosate treatment, which does not stimulate the formation of peroxides. The activities of catalase and dehydroascorbate reductase were increased by the treatment of cold and paraquat, while 3-amino-1,2,4-triazole did not affect the dehydroascorbate reductase activity. The activity of ascorbate free radical reductase increased after treatment by paraquat only. 相似文献
6.
Zapolska-Downar D Zapolska-Downar A Bukowska H Gałka H Naruszewicz M 《Life sciences》1999,65(22):2289-2303
Oxidative modification of LDL by vascular cells has been proposed as the mechanism by which LDL become atherogenic. The effect of ibuprofen on LDL modification by copper ions, monocytes and endothelial cells was studied by measuring lipid peroxidation products. Ibuprofen inhibited LDL oxidation in a dose-dependent manner over a concentration range of 0.1 to 2.0 mM. Ibuprofen (2 mM, 100 microg/ml LDL) reduced the amount of lipid peroxides formed during 2 and 6 h incubation in the presence of copper ions by 52 and 28%, respectively. Weak free radical scavenging activity of ibuprofen was observed in the DPPH test. The protective effect of ibuprofen was more marked when oxidation was induced by monocytes or endothelial cells. Ibuprofen (1 mM, 100 microg/ml LDL) reduced the amount of lipid peroxides generated in LDL during monocyte-mediated oxidation by 40%. HUVEC-mediated oxidation of LDL in the absence and presence of Cu2+ was reduced by 32 and 39%, respectively. More lipid peroxides appeared when endothelial cells were stimulated by IL-1beta or TNFalpha and the inhibitory effect of ibuprofen in this case was more pronounced. Ibuprofen (1 mM, 100 microg/ml LDL) reduced the amount of lipid peroxides formed during incubation of LDL with IL-1beta-stimulated HUVEC by 43%. The figures in the absence and presence of Cu2+ for HUVEC stimulated with TNFalpha were 56 and 59%, respectively. To assess the possibility that ibuprofen acts by lowering the production rate of reactive oxygen species, the intracellular concentration of H2O2 was measured. Ibuprofen (1 mM) reduced intracellular production of hydrogen peroxide in PMA-stimulated mononuclear cells by 69%. When HUVEC were stimulated by IL-1beta or TNFalpha the reduction was 62% and 66%, respectively. 相似文献
7.
DNA damage and lethal effects of hydrogen peroxide and menadione in Chinese hamster cells: distinct mechanisms are involved 总被引:2,自引:0,他引:2
Menadione (Md)-resistant variants of V79 Chinese hamster cells were derived by culturing cells in progressively higher concentrations of this drug. Along with Md resistance these cells acquired cross-resistance to H2O2, which was retained when cells were cultured back in the absence of Md for 18 cell passages. This indicates that some stable alteration is responsible for the modification and may suggest that the toxicity of Md is mediated by oxygen activation. In agreement with the latter a 2.8-fold increase in catalase activity and a 1.5-fold increase in glutathione content were observed in Md-resistant cells as compared to parental cells, whereas superoxide dismutase and glutathione peroxidase remained unaltered. The use of inhibitors of Fenton reaction, inhibitors of enzymatic and nonenzymatic lipid peroxidation and OH radical scavengers, indicated that both DNA damage and cytotoxic effects of H2O2 and Md are mediated by OH radical, without intervention of lipid peroxides. However whereas the catalysis by iron was required for toxicity of extracellular H2O2, it was not involved in Md-induced toxicity. Possible explanations for this difference have been considered, one of them assuming that an excess of the semiquinone form of Md in the cell might replace iron II as a H2O2 reductant, producing OH radical by an organic Fenton reaction. 相似文献
8.
Evidence presented in this report suggests that the hydroxyl radical (OH.), which is generated from liver microsomes is an initiator of NADPH-dependent lipid peroxidation. The conclusions are based on the following observations: 1) hydroxyl radical production in liver microsomes as measured by esr spin-trapping correlates with the extent of NADPH induced microsomal lipid peroxidation as measured by malondialdehyde formation; 2) peroxidative degradation of arachidonic acid in a model OH · generating system, namely, the Fenton reaction takes place readily and is inhibited by thiourea, a potent OH · scavenger, indicating that the hydroxyl radical is capable of initiating lipid peroxidation; 3) trapping of the hydroxyl radical by the spin trap, 5,5-dimethyl-1-pyrroline-1-oxide prevents lipid peroxidation in liver microsomes during NADPH oxidation, and in the model system in the presence of linolenic acid. The possibility that cytochrome P-450 reductase is involved in NADPH-dependent lipid peroxidation is discussed. The optimal pH for the production of the hydroxyl radical in liver microsomes is 7.2. The generation of the hydroxyl radical is correlated with the amount of microsomal protein, possibly NADPH cytochrome P-450 reductase. A critical concentration of EDTA (5 × 10?5m) is required for maximal production of the hydroxyl radical in microsomal lipid peroxidation during NADPH oxidation. High concentrations of Fe2+-EDTA complex equimolar in iron and chelator do not inhibit the production of the hydroxyl radical. The production of the hydroxyl radical in liver microsomes is also promoted by high salt concentrations. Evidence is also presented that OH radical production in microsomes during induced lipid peroxidation occurs primarily via the classic Fenton reaction. 相似文献
9.
V E Kagan O Monovich S R Ribarov 《Biulleten' eksperimental'no? biologii i meditsiny》1985,100(8):179-181
Using the chemiluminescence technique to assay the activity of cholesterol oxidase it has been shown that enzymic oxidation of cholesterol to cholest-4-en-3-one red cell membranes is accompanied by accumulation of lipid peroxidation products--malonyl dialdehyde (MDA). The amount of MDA formed was dependent on the amount of cholesterol oxidized. The free radical scavenger 4-methyl-2,6-ditretbutylphenol, the transition metal chelator EDTA and catalase inhibited lipid peroxidation in red blood cells. The participation of OH radicals in the initiation of lipid peroxidation in red cell membranes in the course of cholesterol oxidation is discussed. 相似文献
10.
Low density lipoprotein cytotoxicity induced by free radical peroxidation of lipid 总被引:40,自引:0,他引:40
Low density lipoprotein (LDL) has been reported to be injurious or toxic to cells in vitro. This injurious effect is, in some instances, due to oxidation of the lipid moiety of the lipoprotein. The objectives of this study were to determine if the oxidation rendering the lipoprotein toxic to human skin fibroblasts occurred by free radical mechanisms, and if so, which of the common free radical oxygen species were involved. The selective free radical blockers or scavengers employed included superoxide dismutase for superoxide, catalase for hydrogen peroxide, dimethylfuran for singlet molecular oxygen, and mannitol for hydroxyl radical. The presence during lipoprotein preparation of general free radical scavengers (vitamin E, butylated hydroxytoluene) or the divalent cation chelator ethylenediamine tetraacetic acid prevented the formation of cytotoxic low density lipoprotein, while the simultaneous presence of superoxide dismutase and catalase partially inhibited its formation. The results indicate that superoxide and/or hydrogen peroxide are involved in the formation of the toxic LDL lipid. The toxic action of oxidized LDL could not be prevented by inclusion of antioxidants in the culture medium, indicating that an oxidized lipid was responsible for cell injury rather than free radicals generated in culture by the action of oxidized LDL. Three separate assays for cell injury (enumeration of attached cells, cell loss of lactate dehydrogenase into the culture medium, and trypan blue uptake) indicated a sequence of events in which the fibroblasts are injured, die, and then detach. 相似文献
11.
The antioxidant enzymes superoxide dismutase (SOD; EC 1.15.1.1), catalase (EC 1.11.1.6), selenium-dependent glutathione peroxidase (SeGPX; EC 1.11.1.9), glutathione reductase (EC 1.6.4.2) and DT-diaphorase (EC 1.6.99.2), plus total GPX activity (sum of SeGPX and Se-independent GPX activities), were studied in 13 500 g supernatants of embryos and 3-day and 11-day post-hatch larvae of turbot Scophthalmus maximus L. SOD activity decreased progressively during development from embryos to 11-day-old larvae, indicative of a decreased need to detoxify superoxide anion radical (O2 − ). In contrast, catalase, SeGPX and glutathione reductase activities increased progressively from embryos to 11-day-old larvae, indicative of an increased need to metabolize hydrogen peroxide (H2 O2 ) and organic peroxides. Consistent with the latter changes, levels of lipid peroxides (i.e. thiobarbituric acid reactive substances) increased 13-fold from embryos to 3-day-old larvae, whilst total peroxidizable lipid was indicated to decrease. Increases were seen for NADPH-dependent DT-diaphorase (after hatching) and total GPX (between 3 and 11 days post-hatch) activities, whilst no change was found in NADH-dependent DT-diaphorase activity. Overall, the results demonstrate a capacity for early life-stages of S. maximus to detoxify reactive oxygen species (O2 − and H2 O2 ) and other pro-oxidant compounds (organic peroxides, redox cycling chemicals). Furthermore, qualitative and quantitative antioxidant changes occur during hatching and development, possibly linked to such events as altered respiration rates (SOD changes) and tissue reorganization and development (catalase, SeGPX, lipid peroxidation). 相似文献
12.
Milczarek R Sokołowska E Hallmann A Kaletha K Klimek J 《The Journal of steroid biochemistry and molecular biology》2008,110(3-5):230-235
During pregnancy placenta is the most significant source of lipid hydroperoxides and other reactive oxygen species (ROS). The increased production of lipid peroxides and other ROS is often linked to pre-eclampsia. It is already proved that placental endoplasmic reticulum may be an important place of lipid peroxides and superoxide radical production. In the present study we revealed that NADPH- and iron-dependent lipid peroxidation in human placental microsomes (HPM) inhibit placental aromatase--a key enzyme of estrogen biosynthesis in human placenta. We showed that significant inhibition of this enzyme is caused by small lipid peroxidation (TBARS (thiobarbituric acid-reactive substances)<4nmol/mg microsomal protein (m.p.)). More intensive lipid peroxidation (TBARS>9nmol/mg microsomal protein) diminishes aromatase activity to value being less than 5% of initial value. NADPH- and iron-dependent lipid peroxidation also causes disappearance of cytochrome P450 parallel to observed aromatase activity inhibition. EDTA, alpha-tocopherol, MgCl(2) and superoxide dismutase (SOD) prevent aromatase activity inhibition and cytochrome P450(AROM) degradation. Mannitol and catalase have not effect on TBARS synthesis, aromatase activity and cytochrome P450 degradation. In view of the above we postulate that the inhibition of aromatase activity observed is mainly a consequence of cytochrome P450(AROM) degradation induced by lipid radicals. The role of hydroxyl radical in cytochrome P450 degradation is negligible in our experimental conditions. The results presented here also suggest that the inhibition of aromatase activity can also take place in placenta at in vivo conditions. 相似文献
13.
Bukowska B 《Cellular & molecular biology letters》2004,9(2):261-270
The effects of the exposure of human erythrocytes to different concentrations of 2,3,7,8-tetrachlorodibenzo-p-dioxin were studied. Particular attention was paid to lipid peroxidation, haemoglobin oxidation, and changes in the activity of catalase and glutathione peroxidase. Human erythrocytes at a 5% haematocrit were incubated with 2,3,7,8-TCDD at concentrations of 0.2 ppm to 1.6 ppm (ng-microg/ml erythrocytes) for 1 hour. The results obtained show that 2,3,7,8-TCDD induces the generation of lipid peroxides and the oxidation of Hb, and decreases the activity of catalase and glutathione peroxidase. This supports the thesis that TCDD causes oxidative stress in erythrocytes. 相似文献
14.
Leoni V Albertini R Passi A Abuja PM Borroni P D'Eril GM De Luca G 《Free radical research》2002,36(5):521-529
Glucose at pathophysiological concentrations was able to accelerate copper-induced oxidation of isolated low-density lipoprotein (LDL) and whole serum. The efficiency of glucose was favored under the following circumstances: (a) when LDL oxidation was induced by low copper concentration, (b) when LDL was partly oxidized, i.e. enriched with lipid peroxides. The glucose derivative methyl-alpha-D-glucoside was ineffective on Cu2+-induced LDL oxidation, pointing out the essential role of the reactivity of the aldehydic carbon for the pro-oxidative effect. When LDL oxidation was induced by a peroxyl radical generator, as a model of transition metal independent oxidation, glucose was ineffective. Glucose was found to stimulate oxidation of LDL induced by ceruloplasmin, the major copper-containing protein of human plasma. Thus, glucose accelerated oxidation of LDL induced by both free and protein bound copper. Considering the requirement for catalytically active copper and for the aldehydic carbon, the pro-oxidative effect of glucose is likely to depend on the increased availability of Cu+; this is more efficient in decomposing lipid peroxide than Cu2+, accounting for acceleration of LDL oxidation. The possible biological relevance of our work is supported by the finding that glucose was able to accelerate oxidation of whole serum, which was assessed by monitoring low-level chemiluminescence associated with lipid peroxidation. 相似文献
15.
Rat liver microsomes were incubated in neutral aqueous solution of potassium peroxychromate, a system which generates singlet molecular oxygen. Such incubation resulted both in a rapid decline in NADPH-cytochrome c reductase activity, and in an increase in formation of lipid peroxides. These reactions were not inhibited by either superoxide dismutase (SOD) or mannitol, nor were they entirely duplicated by incubating microsomes with hydrogen peroxide. However, a high concentration of 1,4-diazabicyclo-[2,2,2]octane (DABCO), a known scavenger of singlet oxygen, prevented both decline in reductase activity and formation of lipid peroxides. These results suggest that the observed effects are, in fact, attributable to singlet oxygen, and not to hydrogen peroxide, superoxide radical, or hydroxyl radical. 相似文献
16.
Germinating seedlings of mung bean ( Phaseolus vulgaris L. cv. K-16) were treated with different concentrations of cadmium acetate (10, 50 and 100 μ M ). Cd2+ lowered the chlorophyll and heme levels. The level of lipid peroxides were higher on day 3 than on day 6. However, Cd2+ treatment significantly enhanced the level of lipid peroxides. Similarly, a dose-dependent induction of lipoxygenase (EC 1.13.11.12) activity was observed with Cd2+ treatment. Further, the activities of antioxidant enzymes such as superoxide dismutase (EC 1.15.1.1) and catalase (EC 1.11.1.6) were decreased. Our results suggest that lipoxygenase-mediated accumulation of lipid peroxides on the one hand and inhibition of free radical scavenging enzymes like superoxide dismutase and catalase on the other caused a pronounced reduction in the chlorophyll and heme levels of the seedlings. The experiments conducted on the effect of Cd2+ on dark-grown seedlings did not conform with the result of light-grown seedlings. Though chlorophyll and heme levels decreased in a dose-dependent manner, no accumulation of lipid peroxides was observed, suggesting that the inhibition of chlorophyll synthesis by Cd2+ is achieved both by reaction with constituent biosynthetic enzymes as well as peroxide-mediated degradation. 相似文献
17.
18.
《Journal of Free Radicals in Biology & Medicine》1986,2(4):267-274
EPR and spin-trapping techniques were employed to study the oxidation of the dihydronicotinamide adenine dinucleotide (NADH) photosensitized by an anthrapyrazole-antitumor agent. The superoxide radical was detected as a DMPO adduct upon illumination of the system with visible light. Photoinduced generation of hydroxyl radicals is demonstrated by detection of DMPO adducts of OH scavengers, such as ethyl alcohol, sodium formate, and sodium azide. The dependence of the production of these spin adducts on the presence of catalase implies the involvement of hydrogen peroxide in that process. The production of hydrogen peroxide is demonstrated independently during oxygen consumption measurements with the Clark electrode technique. 相似文献
19.
Marar T 《Chemico-biological interactions》2011,193(2):149-153
Cells under aerobic condition are always threatened with the insult of reactive oxygen species, which are efficiently taken care of by the highly powerful antioxidant systems of the cell. The erythrocytes (RBCs) are constantly exposed to oxygen and oxidative stress but their metabolic activity is capable of reversing the injury under normal conditions. In vitro hemolysis of RBCs induced by 5, 10 and 20 mM glucose was used as a model to study the free radical induced damage of biological membranes in hyperglycemic conditions and the protection rendered by vitamin E on the same. RBCs are susceptible to oxidative damage, peroxidation of the membrane lipids, release of hemoglobin (hemolysis) and alteration in activity of antioxidant enzymes catalase and superoxide dismutase. The glucose induced oxidative stress and the protective effect of vitamin E on cellular membrane of human RBCs manifested as inhibition of membrane peroxidation and protein oxidation and restoration of activities of superoxide dismutase and catalase, was investigated.Thiobarbituric acid reactive substances are generated from decomposition of lipid peroxides and their determination gives a reliable estimate of the amount of lipid peroxides present in the membrane. Vitamin E at 18 μg/ml (normal serum level) strongly enhanced the RBC resistance to oxidative lysis leading to only 50–55% hemolysis in 24 h, whereas RBCs treated with 10 and 20 mM glucose without vitamin E leads to 70–80% hemolysis in 24 h. Levels of enzymic antioxidants catalase, superoxide dismutase and nonenzymic antioxidants glutathione showed restoration to normal levels in presence of vitamin E. The study shows that vitamin E can protect the erythrocyte membrane exposed to hyperglycemic conditions and so a superior antioxidant status of a diabetic patient may be helpful in retarding the progressive tissue damage seen in chronic diabetic patients. 相似文献
20.
《Free radical research》2013,47(4-5):227-235
It was observed that during the storage of human extracellular fluids at – 20°C the azide-inhibitable ferroxidase activity of caeruloplasmin declined, whilst a new azide-resistant ferroxidase activity (ARFA) developed. The literature suggested that storage-induced ARFA might be due to either a poorly defined enzymatic activity of a low density lipoprotein (LDL) or to lipid peroxides formed within the different lipoprotein fractions. To study this further, the major lipoprotein classes were separated from human serum by density gradient centrifugation. After storage of the lipoprotein fractions, it was found that the LDL fraction had the highest specific activity of ARFA and the highest content of lipid peroxidation products, as assessed by diene conjugates. The ARFA of LDL correlated with its content of diene conjugates and TBA reactive material, which initially suggested that the Fe(II) oxidising activity of peroxidised LDL arose from the reduction of peroxides by Fe(II) in the classical reaction between the metal ion and free radical reduction of lipid peroxides. However. steady state kinetic analysis indicated an enzymic role of LDL in Fe(II) oxidation, with lipid peroxides acting as a substrate for the enzyme. These results indicate that LDL may contain a peroxidase activity. catalysing the oxidation of Fe(II) by lipid peroxides, as well as a ferrous oxidase activity where O2 is the oxidising substrate. 相似文献