Ozone-induced inactivation of antioxidant enzymes

Ozone is an air pollutant that damages a variety of biomolecules. We investigated ozone-induced inactivation of three major antioxidant enzymes. Cu/Zn superoxide dismutase was inactivated by ozone in a concentration-dependent manner. The concentration of ozone for 50% inactivation was approximately 45 microM when 10 microM Cu/Zn superoxide dismutase was incubated for 30 min in the presence of ozone. SDS-polyacrylamide gel electrophoresis (PAGE) showed that the enzyme was randomly fragmented. Both ascorbate and glutathione were very effective in protecting Cu/Zn superoxide dismutase from ozone-induced inactivation. The other two enzymes, catalase and glutathione peroxidase, were much more resistant to ozone than Cu/Zn superoxide dismutase. The ozone concentrations for 50% inactivation of 10 microM catalase and glutathione peroxidase were 500 and 240 microM, respectively. SDS-PAGE demonstrated that ozone caused formation of high molecular weight aggregates in catalase and dimerization in glutathione peroxidase. Glutathione protected catalase and glutathione peroxidase from ozone but the effective concentrations were much higher than that for Cu/Zn superoxide dismutase. Ascorbate was almost ineffective. The result suggests that, among the three antioxidant enzymes, Cu/Zn superoxide dismutase is a major target for ozone-induced inactivation and both glutathione and ascorbate are very effective in protecting the enzyme from ozone.     

Effects of Lead on Hepatic Antioxidant Status and Transcription of Superoxide Dismutase Gene in Pigs

Ninety-six castrated boars (Duroc x Landrace x Yorkshire) were randomly divided into four groups, each of which was replicated three times with eight pigs. The groups received the same basal diet supplemented with 0, 5, 10, and 20 mg/kg lead, respectively. The malondialdehyde and glutathione levels, antioxidant enzymes activities, and zinc/copper superoxide dismutase (Zn/Cu SOD) mRNA content in the liver were determined to evaluate the lead hepatic intoxication caused by the lead. Results showed the increased lipid peroxides level and the reduced glutathione content, along with a concomitant decrease in the activities of superoxide dismutase, catalase, and glutathione peroxidase. Moreover, the level of hepatic Zn/Cu SOD mRNA was also significantly reduced. We suggest potential mechanism for lead intoxication in liver as follows: lead causes parallel decrease in Zn/Cu SOD mRNA and activities of antioxidant enzymes, leading to the declined ability of scavenging free radicals with excessive production of lipid peroxides, which seriously damages the hepatic structure and function.     

Response of the antioxidant defense system to tert-butyl hydroperoxide and hydrogen peroxide in a human hepatoma cell line (HepG2)

The aim of this work was to investigate the response of the antioxidant defense system to two oxidative stressors, hydrogen peroxide and tert-butyl hydroperoxide, in HepG2 cells in culture. The parameters evaluated included enzyme activity and gene expression of superoxide dismutase, catalase, glutathione peroxidase, and activity of glutathione reductase. Besides, markers of the cell damage and oxidative stress evoked by the stressors such as cell viability, intracellular reactive oxygen species generation, malondialdehyde levels, and reduced glutathione concentration were evaluated. Both stressors, hydrogen peroxide and tert-butyl hydroperoxide, enhanced cell damage and reactive oxygen species generation at doses above 50 microM. The concentration of reduced glutathione decreased, and levels of malondialdehyde and activity of the antioxidant enzymes consistently increased only when HepG2 cells were treated with tert-butyl hydroperoxide but not when hydrogen peroxide was used. A slight increase in the gene expression of Cu/Zn superoxide dismutase and catalase with 500 microM tert-butyl hydroperoxide and of catalase with 200 microM hydrogen peroxide was observed. The response of the components of the antioxidant defense system evaluated in this study indicates that tert-butyl hydroperoxide evokes a consistent cellular stress in HepG2.     

Effects of fluoride on hepatic antioxidant system and transcription of Cu/Zn SOD gene in young pigs

Thirty-two barrows (Duroc x Landrace x Yorkshire) were randomly divided into four groups, each of which included eight pigs. The groups received the same basal diet supplemented with 0, 100, 250 and 400mg/kg fluoride, respectively. The malondialdehyde (MDA) and glutathione (GSH) levels, antioxidant enzymes activities and zinc/copper superoxide dismutase (Cu/Zn SOD) mRNA content in the liver were determined to evaluate the fluoride hepatic intoxication. Results showed the increased lipid peroxides (LPO) level and the reduced GSH content, along with a concomitant decrease in the activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px). Moreover, the level of hepatic Cu/Zn SOD mRNA was also significantly reduced. We suggest the mechanism of fluoride injuring the liver as follows: fluoride causes a decrease in Cu/Zn SOD mRNA and the reduced activities of antioxidant enzymes, leads to the declined ability of scavenging free radicals with excessive production of LPO, which seriously damages the hepatic structure and function.     

Ontogenic characteristics of lipid peroxidation and antioxidant system enzymes in the brain of geese

Peculiarities of antioxidant homeostasis of geese brain tissue during embryogenesis and early postnatal period have been studied. It has been shown that the cerebrum and hindbrain tissues are characterized by a higher level of lipid peroxidation compared to liver. Main antioxidative enzymes' activity (superoxide dismutase, catalase, glutathione peroxidase) in the brain already reaches its maximum in the middle period of embryogenesis. We have found that brain tissues are characterized by a lower activity of intracellular enzymes (superoxide dismutase, catalase) but increased glutathione peroxidase activity as compared to liver. The rate of Fe2+ initialized lipid peroxidation and coefficient of antioxidative activity were used as a criterion for evaluation of antioxidative system's status. According to the dynamics of these factors the highest tension of antioxidative system in the brain appears in the period of the contour (28 days) and juvenile (49 days) feather formation.     

Antioxidative signalling pathways regulate the level of reactive oxygen species at the endometrial-extraembryonic membranes interface during early pregnancy

Conceptus (embryo and associated extraembryonic membranes) implantation and development require a reciprocal biochemical and physical interactions between the extraembryonic membranes and the endometrium. However, the enzymatic antioxidative pathways controlling reactive oxygen species production at the endometrial-extraembryonic membrane interface early in pregnancy are not known. We aimed therefore to determine the content of malondialdehyde, as biomarkers of lipid peroxidation, and the activities of the major antioxidant enzymes, copper-zinc containing and manganese containing superoxide dismutases, catalase and glutathione peroxidase, in sheep extraembryonic membranes, caruncular and intercaruncular endometrium zones sampled at specific stages of pregnancy corresponding to the conceptus implantation (day 16) and the early post-implantation period (day 21). Malondialdehyde content in caruncular, intercaruncular and extraembryonic tissues was not different between stages of the pregnancy. Extraembryonic membranes demonstrated increased manganese containing superoxide dismutase and glutathione peroxidase activities, whereas catalase activity in these tissues decreased from day 16 to day 21. Caruncular tissues demonstrated increased manganese containing superoxide dismutase activity from day 16 to day 21. Intercaruncular tissues demonstrated increased copper-zinc containing superoxide dismutase, manganese containing superoxide dismutase and catalase activities from day 16 to day 21. The ovine extraembryonic membranes exhibit dynamic changes in enzymatic antioxidative pathways different from those of endometrial tissues during the transition from implantation to post-implantation period. This biochemical data provides novel insights into the developmental changes in antioxidative pathways of extraembryonic membranes and endometrium during early conceptus development.     

Zinc-alleviating effects on iron-induced phytotoxicity in roots of <Emphasis Type="Italic">Triticum aestivum</Emphasis>

The mechanisms of growth inhibition and antioxidative response were investigated in wheat roots exposed to 300 μM iron together with different zinc concentrations (0, 50, and 250 μM). All Zn concentrations decreased Fe content but increased Zn content in the roots and leaves of Fe-treated seedlings. Compared with Fe stress alone, 50 or 250 μM Zn + Fe treatment stimulated root growth, and increased cell viability but decreased malondialdehyde content, which were correlated with the decreases of total and apoplastic hydrogen peroxide and superoxide anion radical (O₂ ^·?) content along with apoplastic hydroxyl radical content. Generation of O₂ ^·? in response to 10 μM diphenylene iodonium suggested that NADPH oxidase activity was lower in Zn + Fe-treated roots than in other roots. In addition, cell wallbound peroxidase, diamine oxidase, and polyamine oxidase in Fe-treated roots were insensitive to Zn addition. Further study showed the stimulation of total superoxide dismutase and glutathione reductase (GR) activities as well as apoplastic catalase, ascorbate peroxidase, and GR in Zn + Fe-stressed roots in comparison with Fe-alone-treated ones. Taken together, Zn could alleviate iron-inhibitory effect on root growth, which might be associated with the decrease of lipid peroxidation, the increase of cell viability and the reductions of reactive oxygen species generation.     

Antioxidant enzymatic systems in neuronal and glial cell-enriched fractions of rat brain during aging

The activities of Cu,Zn superoxide dismutase, glutathione peroxidase, catalase and glutathione reductase in neuronal and glial cell-enriched fractions obtained from the cerebral cortex of rat brain during aging (15, 30, 90, 350, 750 days of age) were assayed. Our results showed that glutathione peroxidase, catalase and glutathione reductase activities varied little during the examined periods. Only the Cu,Zn superoxide dismutase activity decreased notably from 15th to 750th day of age in both neuronal and glial cells, moreover the activities of all enzymes studied were always detected at lower levels in neuronal cells with respect to glial cells. In agreement with diminished SOD activity, the lipid peroxidation showed an elevated increase with aging; this fact is more evident in neuronal than in glial cells. In conclusion our data show that Cu,Zn superoxide dismutase is the most affected antioxidant enzymatic system of brain aging and it could be responsible for the increased lipid peroxidation in both cell types examined.A preliminary report of these results was presented at the 19th Meeting F.E.B.S. Rome July 2–7, 1989.     

Cadmium and copper induction of oxidative stress and antioxidative response in tomato (<Emphasis Type="Italic">Solanum lycopersicon</Emphasis>) leaves

We compare cadmium and copper induced oxidative stress in tomato leaves and the antioxidative enzyme response during a time course of 96 h. Plants were subjected to 25 μM of CdCl₂ or CuSO₄ and malondialdehyde (MDA) level and activity of guaiacol peroxidase, superoxide dismutase, catalase, ascorbate peroxidase and glutathione reductase were determined. The results showed that there was an early increase in the MDA level and in the guaiacol peroxidase activity more pronounced with copper exposure during almost all the time course of the experiment. The activity of superoxide dismutase and catalase was induced very early after cadmium and copper treatment, reached a maximal value after 12 h and then declined but it remained always slightly higher than the control at the end of the experiment. Ascorbate peroxidase activity pathway was similar to superoxide dismutase or catalase with a maximal activity after 48 h of heavy metal exposure. Induction of glutathione reductase activity observed only under copper exposure is maintained during almost all the experimental time. The antioxidative activity developed by tomato leaves is more induced by copper treatment. This can be related to the ability of this metal to induce more than cadmium an accumulation of reactive oxygen species (ROS) at the cellular level. Decline in the antioxidative enzymes activity at the end of the experiment can be a consequence of cadmium- and copper-inducing a further ROS formation that might affect enzymes activity.     

光质对烟草叶片生长发育过程中抗氧化系统的影响

以云烟87植株为材料,通过覆盖白、红、黄、蓝、紫色滤膜获得不同光质,于大田条件下研究了光质对烟草叶片生长发育过程中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、过氧化物酶(POD)、抗坏血酸过氧化物酶(APX)、谷胱甘肽过氧化物酶(GPX)、谷胱甘肽还原酶(GR)等抗氧化酶活性,抗氧化剂谷胱甘肽(GSH)和抗坏血酸(AsA)以及丙二醛(MDA)含量的影响.结果表明,在烟草植株第11片叶生长发育的7~70 d内,其抗氧化酶活性和抗氧化物质含量呈现先升高后下降的变化趋势.与白光(对照)相比,黄光诱导烟草叶片SOD、CAT、APX和GR活性升高,以及AsA和GSH含量增加;而红光诱导APX和GR活性上升,以及GSH和AsA含量升高;但紫光却使SOD、CAT、POD、GR和GPX活性下降,GSH和AsA含量降低,而蓝光则使所有抗氧化酶活性和抗氧化物质含量降低.紫光和蓝光处理的烟草叶片中MDA含量较高,而黄光和红光处理的则较低.总体而言,在大田条件下,相对红光和黄光而言,蓝光和紫光处理下的烟草叶片更容易发生光氧化胁迫.     

Protection against radical peroxidations in cerebral aging in cerebral capillaries and microvessels]

Systemic injection of substances that generate free radicals into the rat induces deleterious alteration of various tissues (as demonstrated by the production of conjugated dienes and malondialdehyde) but only has a slight effect on the brain. This shows that the blood-brain barrier has important protective properties. In fact the cerebral capillaries and microvessels have superoxide dismutase, glutathione peroxidase, and catalase activities higher than those found in the rest of the cerebral tissue during aging. These activities vary little, except for catalase which decreases. But, curiously, the concentrations of Mn, Cu, and Zn are not related to enzymatic activity, although the micro-elements are necessary for the activity of superoxide dismutase. On the other hand, during aging, the capillaries and cerebral microvessels undergo extensive modifications at the level of the polyunsaturated fatty acids: for example, the concentration of arachidonic acid decreases by half.     

Dietary Zinc Glycine Chelate on Growth Performance,Tissue Mineral Concentrations,and Serum Enzyme Activity in Weanling Piglets

One hundred twenty crossbred piglets (Duroc × Landrace × Yorkshire) were used to determine the effects of dietary zinc glycine chelate on growth performance, tissue mineral concentrations, and serum enzyme activity. All pigs were allotted to four treatments and fed with basal diets supplemented with 0, 50, and 100 mg/kg Zn as zinc glycine chelate or 3,000 mg/kg Zn as zinc oxide (ZnO). After the 35-day feeding trial, results of the study showed that, compared to the control, average daily gain was improved (P < 0.05) for pigs fed 100 mg/kg Zn from zinc glycine chelate or 3,000 mg/kg Zn from ZnO and Zn concentrations in serum and M. longissimus dorsi were significantly enhanced by 100 mg/kg dietary zinc glycine chelate and 3,000 mg/kg ZnO. In addition, supplementation of 100 mg/kg zinc glycine chelate decreased (P < 0.05) the liver Fe level, liver Zn level, spleen Cu level, and kidney Cu level compared to that of the 3,000-mg/kg ZnO group. For feces mineral excretion, 3,000 mg/kg Zn from ZnO greatly increased the concentration of fecal Zn (P < 0.01) and Mn (P < 0.05) compared to that of the control or the 100-mg/kg zinc glycine chelate group. Moreover, alkaline phosphatase and Cu/Zn superoxide dismutase activities of pigs in 100 mg/kg zinc glycine chelate and ZnO treatments were greatly higher than that of the control. The results of present study showed that supplementation with zinc glycine chelate could improve growth and serum enzyme activities and could also decrease zinc excretion in feces in weanling pig compared to high dietary ZnO.     

Effects of Cadmium,Chromium and Lead on Growth,Metal Uptake and Antioxidative Capacity in <Emphasis Type="Italic">Typha angustifolia</Emphasis>

This study investigates the modulation of antioxidant defence system of Typha angustifolia after 30 days exposure of 1 mM chromium (Cr), cadmium (Cd), or lead (Pb). T. angustifolia showed high tolerance to heavy metal toxicity with no visual toxic symptom when exposed to metal stress, and Cd/Pb addition also increased plant height and biomass especially in Pb treatment. Along with increased Cr, Cd, and Pb uptake in metal treatments, there was enhanced uptake of plant nutrients including Ca and Fe, and Zn in Pb treatment. A significant increase in malondialdehyde (MDA) content and superoxide dismutase (SOD) and peroxidase (POD) activities were recorded in plants subjected to Cr, Cd, or Pb stress. Furthermore, Pb stress also improved catalase (CAT), ascorbate peroxidase (APX), and glutathione peroxidase (GPX) activities; whereas Cr stress depressed APX and GPX. The results indicate that enzymatic antioxidants and Ca/Fe uptake were important for heavy metal detoxification in T. angustifolia, stimulated antioxidative enzymes, and Ca, Fe, and Zn uptake could partially explain its hyper-Pb tolerance.     

Influences of magnesium deficiency and cerium on antioxidant system of spinach chloroplasts

Magnesium-deficiency conditions applied to spinach cultures caused an oxidative stress status in spinach chloroplast monitored by an increase in reactive oxygen species (ROS) accumulation. The enhancement of lipids peroxide of spinach chloroplast grown in magnesium-deficiency media suggested an oxidative attack that was activated by a reduction of antioxidative defense mechanism measured by analysing the activities of superoxide dismutase, catalase, ascorbate peroxidase, guaiacol peroxidase, and glutathione reductase, as well as antioxidants such as carotenoids and glutathione content. As the antioxidative response of chloroplast was reduced in spinach grown in magnesium-deficiency media, it caused a significant reduction of spinach plant weight, old leaves turning chlorosis. However, cerium treatment grown in magnesium-deficiency conditions decreased the malondialdehyde and ROS, and increased activities of the antioxidative defense system, and improved spinach growth. Together, the experimental study implied that cerium could partly substitute for magnesium and increase the oxidative stress-resistance of spinach chloroplast grown in magnesium-deficiency conditions, but the mechanisms need further study.     

Effect of Sex and Dietary Organic Zinc on Growth Performance, Carcass Traits, Tissue Mineral Content, and Blood Parameters of Broiler Chickens

Zinc (Zn) is an essential mineral for animal development and function. A study was carried out to evaluate the effect of sex and dietary organic zinc (OZ) on growth performance, carcass traits, tissue mineral content, and blood parameters of broiler chickens. A total of 240 1-day-old male and 240 female broiler chicks (Cobb × Cobb) were assigned to two dietary levels of OZ (2 × 2 factorial) with six replicates per treatment (20 birds/replicate pen). The OZ supplementation levels were 0 and 25 ppm. Results showed that OZ supplementation did not affect the growth performance of male and female broilers, but the males showed significantly better (P < 0.05) growth performance than females did. Similarly, OZ supplementation did not affect the thickness of both the back and thigh skin of male and female broilers; however, males had thicker skin than females. Dietary OZ supplementation did not affect collagen contents in the skin and meat samples. Male broilers had higher skin collagen contents than females, but no sex difference was found in meat collagen contents. OZ supplementation did not affect the shear force values of skin and meat samples. Male broilers had higher shear force values of back skin than females, but not in the meat samples. Dietary OZ supplementation increased (P < 0.05) the thigh meat Zn content in both sexes. The plasma Ca content was significantly (P < 0.05) increased by dietary OZ supplementation; however, other blood parameters were not affected by dietary OZ supplementation. Males had higher plasma glucose and cholesterol content than females. It is concluded that dietary OZ supplementation at the level of 25 ppm does not affect the growth performance and skin quality of broiler chickens but increases the Zn content in thigh meat and Ca content in plasma of broiler chickens. Male broilers had better growth performance and skin quality than females.     

Circadian rhythms of biomarkers of oxidative stress and their characteristics in broiler chickens reared under natural light/dark cycle

For the purpose of studying circadian rhythms of biomarkers of oxidative stress in broiler chickens maintained under natural photoperiod, we collected blood from the wing veins of 10 matured broiler chickens every three hours for a period of 24 h. The blood samples were analyzed for superoxide dismutase (SOD), catalase (CAT), and malondialdehyde (MDA). The fitting in of single cosinor procedure showed that CAT and MDA exhibited clear circadian rhythms, while SOD did not. The amplitude of daily rhythms for the parameters was small, while the acrophases were all restricted to the light phase of the light:dark cycle. In conclusion, to our knowledge, this is the first time investigation of the circadian rhythms of antioxidants in broilers chickens, reared under natural photoperiod in the tropics. The result may be exploited for a more precise targeting of reactive oxygen species, thereby offering better protection for the cells to combat oxidative stress.     

Antidiabetic and Pancreas-Protective Effects of Zinc Threoninate Chelate in Diabetic Rats may be Associated with its Antioxidative Stress Ability

Zinc exerts a wide range of important biological roles. The present study was carried out to investigate the effects of zinc threoninate chelate in blood glucose levels, lipid peroxidation, activities of antioxidant defense systems and nitrite concentration, and histology of the pancreas in diabetic rats. Wistar rats were intravenously injected with a single dose of streptozotocin to induce diabetes. Then, diabetic rats were administrated orally with zinc threoninate chelate (3, 6, and 9 mg/kg body weight) once daily for 7 weeks. Fasting blood glucose was monitored weekly. At the end of the experimental period, the diabetic rats were killed, and levels of serum insulin, malondialdehyde, and nitric oxide, activities of glutathione peroxidase, total superoxide dismutase, copper/zinc-superoxide dismutase, and nitric oxide synthase were determined; pancreas was examined histopathologically as well. Zinc threoninate chelate significantly reduced the blood glucose levels and significantly increased the serum insulin levels in diabetic rats. In addition, zinc threoninate chelate caused a significant increase in activities of antioxidant enzymes and significant decrease in nitrite concentration and malondialdehyde formation in the pancreas and serum of diabetic rats. These biochemical observations were supplemented by histopathological examination of the pancreas. These results suggested that the antidiabetic effect of zinc threoninate chelate may be related to its antioxidative stress ability in diabetic rats.     

Correlation between superoxide dismutase,glutathione peroxidase and catalase in isolated rat hepatocytes during fetal development

Superoxide dismutase, glutathione peroxidase and catalase activities were determined in isolated fetal rat hepatocytes of various ages and compared with the values of neonatal and adult cells. The developmental pattern of superoxide dismutase and glutathione peroxidase were very similar with a low constant activity in the fetal cells and a postnatal burst. On the contrary catalase begins to increase already since the 18th day of the fetal life. The results suggest a functional correlation of superoxide dismutase and glutathione peroxidase in the antioxidative enzyme defense of liver cells.     

Combined Effects of Muscular Dystrophy,Ecological Stress,and Selenium on Blood Antioxidant Status in Broiler Chickens

The results obtained in this study demonstrated that experimentally induced alimentary muscular dystrophy (MD) in Cobb 500 broiler chickens resulted in increased plasma concentrations of malondialdehyde (MDA), deviations in activities of erythrocyte antioxidant enzymes Cu,Zn-SOD (decrease), and CAT (increase) as well as reduction in plasma concentrations of trace elements Cu, Zn, and Se in affected birds. These data evidenced the presence of oxidative stress in birds with MD, reared both under conditions of ecological comfort and ecological stress. The increased MDA and САТ levels and the reduced Cu,Zn-SOD, Cu, Zn, and Se concentrations in healthy chickens reared under unfavorable microclimatic conditions such as higher air temperature and humidity, higher ammonia concentrations, and lower light intensity were indicative about an induced ecological stress. After the 10-day oral treatment with a selenium-containing preparation, the levels of MDA, Cu,Zn-SOD, CAT, Cu, Zn, and Se attained their normal values in chickens with MD, reared under ecologically comfortable conditions. According to our results, ecological stress was shown to exert independently a significant adverse effect upon the levels of the studied parameters and possibly to be a cause for their slower and not complete normalization despite the selenium therapy in experimental broiler chickens.     

Superoxide dismutase during glucose repression of Hansenula polymorpha CBS 4732

Hansenula polymorpha CBS 4732 was studied during cultivation on methanol and different glucose concentrations. Activities of Cu/Zn and Mn superoxide dismutase, catalase and methanol oxidase were investigated. During cultivation on methanol, increased superoxide dismutase and catalase activities and an induced methanol oxidase were achieved. Transfer of a methanol grown culture to medium with a high glucose concentration caused growth inhibition, low consumption of carbon, nitrogen and phosphate substrates, methanol oxidase inactivation as well as decrease of catalase activity (21.8 +/- 0.61 deltaE240 x min(-1) x mg protein(-1)). At the same time, a high value for superoxide dismutase enzyme was found (42.9 +/- 0.98 U x mg protein(-1), 25% of which was represented by Mn superoxide dismutase and 75% - by the Cu/Zn type). During derepression methanol oxidase was negligible (0.005 +/- 0.0001 U x mg protein(-1)), catalase tended to be the same as in the repressed culture, while superoxide dismutase activity increased considerably (63.67 +/- 1.72 U x mg protein(-1), 69% belonging to the Cu/Zn containing enzyme). Apparently, the cycle of growth inhibition and reactivation of Hansenula polymorpha CBS 4732 cells is strongly connected with the activity of the enzyme superoxide dismutase.