Tag Archives: antioxidant enzymes

Effects of L-glutamic acid and pyridoxine on glutathione depletion and lipid peroxidation generated by epinephrine-induced stress in rats

N. O. Salyha

Institute of Animal Biology, NAAS of Ukraine, Lviv;
е-mail: ynosyt@yahoo.com

The main goal of this research was to investigate and compare the protective effects of L-glutamic acid (L-Glu) alone and combined with Pyridoxine (L-Glu+Pyridoxine) for the purpose of suppression and mitigation of epinephrine-induced stress in rats. This study outlines possible links between changes of reduced glutathione (GSH) level, antioxidant enzymes activity and content of the lipid peroxidation products after administration of the above-mentioned substances and under the action of stress in various tissues of rats. The obtained results suggest that the GSH level was significantly inhibited by stress in all investigated tissues (except kidneys). We have shown that under the stress, activities of glutathione-associated enzymes were changed (mainly decreased) in all investigated tissues. In rats, additionally received L-Glu and L-Glu+Pyridoxine, much less changes or lack of changes in studied parameters were observed. The content of lipid peroxidation products (lipid peroxides (LOOH) and thiobarbituric acid reactive substances (TBARS)) in myocardium, liver and kidney tissues of experimental groups under the stress conditions were significantly higher compared to the control. While in experimental groups that received L-Glu and L-Glu+Pyridoxine LOOH content in kidney, spleen and liver and TBARS content in spleen, liver and myocardium were almost at the level of control values. These results indicate that L-Glu and L-Glu+Pyridoxine can mitigate and suppress epinephrine-induced stress in rats.

Nitrate and nitrite in drinking water affect antioxidant enzymes in erythrocytes of rats

E. Sierra-Campos1, M. A. Valdez-Solana1, M. I. Campos-Almazán1,
C. Avitia-Domínguez2, J. L. Hernández-Rivera1, J. A. De Lira-Sánchez1,
G. Garcia-Arenas3, A. Téllez-Valencia2

1Faculty of Chemical Sciences, Campus Gómez Palacio, University Juarez of Durango State, Gomez Palacio, Durango, Mexico;
e-mail: ericksier@gmail.com;
2Faculty of Medicine and Nutrition, Campus Durango, University Juarez of Durango State, Durango, Mexico;
3Faculty of Health Sciences, Campus Gómez Palacio, University Juarez of Durango State, Gomez Palacio, Durango, Mexico

The present study evaluated the effect of short term intake of nitrite and nitrate drinking water on the antioxidant system and membrane damage of rat erythrocytes. Wistar rats were randomly divided into three groups as follows; the group I received only distilled water ad libitum; the group II was given water with nitrate (a dose of 124 mg/kg of nitrate-nitrogen) as drinking water and the group III was given nitrites dissolved in distilled water in a dose of 150 mg/kg for 7 days. At the end of the study, group III rats showed a significant decrease in activities of glutathione peroxidase (GPx), glucose 6-phosphate dehydrogenase (G6PDH) and catalase (CAT), while in group II rats, the activity of GPx and CAT were significantly reduced, but no significant changes in glutathione reductase activity and peroxynitrite levels were observed. On the other hand, malondialdehyde (MDA) was increased in both groups with respect to group I. Also, our major results indicate that all treatments changed methemoglobin levels and osmotic fragility in comparison to group I rats. The intensity of alterations was found more severe in rats of group III, followed by rats of group II. It can be concluded from these observations that nitrate or nitrite leads to alterations in the erythrocytes antioxidant defense status mainly throughout NADPH relate enzymes.

Effect of sodium nitroprusside and S-nitrosoglutathione on pigment content and antioxidant system of tocopherol-deficient plants of Arabidopsis thaliana

N. M. Semchuk, Yu. V. Vasylyk, O. I. Kubrak, V. I. Lushchak

Vassyl Stefanyk Precarpathian National University, Ivano-Frankivsk, Ukraine;
e-mail: lushchak@pu.if.ua

Sodium nitroprusside (SNP) and S-nitrosoglutathione (GSNO) were used as a source of exogenous nitric oxide (NO) to investigate their effects on biochemical parameters and antioxidant enzyme response in leaves of wild type Columbia and tocopherol-deficient vte4 and vte1 mutant lines of Arabidopsis thaliana plants and possible tocopherol involvement in regulation of antioxidant response under NO-induced stress. SNP enhanced the activity of the enzymes, that scavenge hydrogen peroxide in leaves of all studied lines, and increased glutathione reductase and glutathione-S-transferase activity there. In addition, it decreased the intensity of lipid peroxidation in vte1 mutant line leaves. At the same time, GSNO increased the levels of protein carbonyls and inactivated enzymes ascorbate peroxidase, guaiacol peroxidase and dehydroascorbate reductase in almost all investigated plant lines. In contrast to wild type, GSNO increased superoxide dismutase activity and decreased catalase activity and chlorophyll a/b ratio in the leaves of two mutant lines. It can be assumed that tocopherols in some way are responsible for plant protection against NO-induced stress. However the mechanisms of this protection remain unknown.

Effect of high sodium chloride concentrations on the pigment content and free-radical processes in corn seedlings leaves

Y. V. Vasylyk, V. I. Lushchak

Vassyl Stefanyk Precarpathian National University, Ivano-Frasnkivk, Ukraine;
e-mail: lushchak@pu.if.ua

The effect of sodium chloride on general morphometrical parameters of seedlings, and biochemical parameters in the leaves of corn seedlings was studied. Exposure to 100 and 200 mM NaCl slowed down the growth of stem and roots, whereas 100 and 200 mM NaCl during 24 h enhanced the concentration of chlorophylls, carotenoids, anthocyans, and thiobarbituric acid reactive substances. The decrease in protein carbonyl groups was found at 24-hour exposure to 200 mM salt. The treatment during 24, 48 and 72 h to 200 mM salt increased the level of total and high molecular mass thiols, whereas low molecular mass thiol content was by 20-25% higher at 48 h exposure to all used salt concentrations. The activity of guaiacol peroxidase was higher only at 24 h exposure to 100 and 200 mM salt, and catalase – at 50 mM during 48 h. At 72-hour exposure, catalase activity was by 27 and 41% higher in seedlings, exposed to 50 and 200 mM NaCl, respectively. Therefore, it is concluded the plant exposure to 50-200 mM salt initially developed oxidative stress, inducing adaptive response – an increase in antioxidant potential and efficiency of systems of energy production. That results in plant adaptation to unfavourable conditions.

Effect of short-term salt stress on oxidative stress markers and antioxidant enzymes activity in tocopherol-deficient Arabidopsis thaliana plants

N. M. Semchuk, Yu. V. Vasylyk, Ok. V. Lushchak, V. I. Lushchak

Vasyl Stefanyk Precarpathian National University, Ivano-Frankivsk, Ukraine;
e-mail: lushchak@pu.if.ua

Changes of carotenoids and anthocyanins content, lipid peroxidation, and activity of antioxidant enzymes were studied in wild type and tocopherol-deficient lines vte1 and vte4 of Arabidopsis thaliana subjected to 200 mM NaCl during 24 h. The salt stress enhanced the intensity of lipid peroxidation to different extent in all three plant lines. Salt stress resulted in an increase of carotenoid content and activity of catalase, ascorbate peroxidase, guaiacol peroxidase and glutathione reductase in wild type and tocopherol-deficient vte1 mutant. However, the increase in anthocyanins concentration was observed in vte1 mutants only. In vte4 mutant, which contain γ-tocopherol instead of α-tocopherol, the response to salt stress occurred via coordinative action of superoxide dismutase and enzymes of ascorbate-glutathione cycle, in particular, ascorbate peroxidase, glutathione reductase, dehydroascorbate reductase, and glutathione-S-transferase. It can be concluded, that salt stress was accompanied by oxidative stress in three studied lines, however different mechanisms involved in adaptation of wild type and tocopherol-deficient lines to salt stress.

Effect of sodium chloride and nitroprusside on protein carbonyl groups content and antioxidant enzyme activity in leaves of corn seedlings Zea mays L.

Yu. V. Vasylyk1, N. M. Semchuk1, Ok. V. Lushchak2, V. I. Lushchak1

1Department of Biochemistry and Biotechnology,
2Botanical Gardens of Vasyl Stefanyk Precarpathian National University,
Ivano-Frankivsk, Ukraine;
e-mail: lushchak@pu.if.ua

The effect of sodium nitroprusside (SNP) and sodium chloride (NaCl) on protein carbonyl group content and activity of antioxidant enzymes was investigated in leaves of maize seedlings. Incubation with NaCl and SNP+NaCl increased the content of carbonyl proteins after 24 h. Treatment with SNP+NaCl during 48 h showed lower and after 72 h higher carbonyl protein content than that in the control. Catalase activity was higher in the leaves of SNP+NaCl-treated than in the leaves of SNP-treated seedlings after 24 h. Ascorbate peroxidase activity increased after incubation with 0.2 mM SNP for 24 h. Significant increment of guaiacol peroxidase activity was obtained in all treated groups in comparison with the control after 72 h. Glutathione-S-transferase activity increased after 48 h seedling treatment with NaCl or SNP and 72 h seedling incubation with NaCl. Under experimental conditions used, glutathione reductase activity was virtually not affected. It is proposed that SNP can be used to prevent salt-induced oxidative stress in maize.

Generation of active oxygen forms in rat tymocytes under action of hydrogen peroxide and fullerene C(60)

S. M. Grebinyk, I. I. Grynyuk, S. V. Prylutska, O. P. Matyshevska

Taras Shevchenko Kyiv National University, Ukraine;
e-mail: grebnik_z@yahoo.com

The dynamics of active oxygen forms (AOF) generation in rat thymocytes 50 min after treatment with 0.1 and 0.5 mM H2O2 was estimated with the use of fluorescent probe DCFDA. Both enhanced AOF generation, which was dependent on H2O2 concentration, and glutathione peroxidase and superoxide dismutase activation, followed by a decrease of thymocytes viability were demonstrated.
Preincubation of cells with 10-5 M fullerene C60 was shown not only to prevent H2O2 – induced AOF generation but to increase viability of H2O2-treated thymocytes at more prolonged time period. The data obtained indicate to fullerene C60 ability to prevent oxidative stress in thymocytes.

Alterations of prooxidant-antioxidant system of rat liver at ethanol and tetracycline action

Kh. Yu. Nedoshytko

I. Ya. Horbachevsky Ternopil State Medical University, Ukraine;
e-mail: khrystynan@ukr.net

The state of antioxidant system and fatty acid composition of lipids in the liver tissues of rats of different sex at the ethanol and tetracycline action and at the influence of biologically active additives (BAA) “Alpha + Omega” at a dose of 0.5 mg/kg b.w. per os was investigated. It was found that the contet of lipid peroxidation products in the liver was increased at the action of 40 % ethanol at a dose of 7 ml/kg b.w. per os and tetracycline – 500 mg/kg and more profound at their joint using. Howe­ver, the content of diene conjugates was stronger increased in the liver of females at the action of ethanol, while in the liver of males at the action of tetracycline (Р < 0.05). It was shown that the application of the investigated compounds led to the reduction of an antioxidant defense system activity of males and females liver, as evidenced by the decrease of superoxide dismutase activity by 46 and 43% and reduction of glutathione content by 39 and 38% (Р < 0.05). The activity of alanineaminotransferase, aspartateaminotransferase and alkalinephosphatase was increased in the liver of males and females under the influence of ethanol and tetracycline and more profound at their joint usa­ge (Р < 0.05). It was established that ethanol and tetracycline unidirectionally changed fatty acid composition of total lipids of rat liver, but at the ethanol action the changes were more expressed in females while at the tetracycline action in males. The application during 14 days of BAA “Alpha + Omega” to male and female rats with an acute tetracycline damage at subacute ethanol action led to partial normalization of prooxidant-antioxidant system and the relative content of total lipids fatty acids of the liver of both sexes animals.

The effect of N-stearoylethanolamine on the activity of antioxidant enzymes, content of lipid peroxidation products and nitric oxide in the blood plasma and liver of rats with induced insulin-resistance

O. V. Onopchenko, G. V. Kosiakova, T. M. Goridko,
A. G. Berdyschev, O. F. Meged, N. M. Hula

Palladin Institute of Biochemistry, National Academy of Science of Ukraine, Kyiv;
e-mail: onop.89.av@mail.ru

The influence of N-stearoylethanolamine (NSE) on the content of lipid peroxidation products­, activity of antioxidant enzymes and the nitric oxide level in the liver and blood plasma of rats with insulin-resistance(IR) state was investigated. IR state was induced in rats by prolonged high-fat diet (58% of energy derived from fat) for 6 months combined with one injection of streptozotocin (15 mg/kg of body weight). The existence of IR state was estimated by results of glucosotolerance test and blood plasma insulin content.  The level of lipid peroxides products was shown to be higher in the liver of insulin resistant animals as a result of reduced superoxide dismutase and catalase activity, however, glutathione peroxidase activity was increased. The increase of nitric-oxide content in the liver and blood plasma of high-fat diet rats compared with healthy control animals was also observed. The administration of the NSE suspension per os in a dose of 50 mg/kg during 2 weeks to the rats with induced insulin-resistance state contributed to the increase of superoxide dismutase, catalase and glutathione peroxidase activi­ty. In consequence of antioxidant enzymes activation the intensity of POL process was decreased. The NSE administration caused normalization of nitric oxide level, restoring pro-/antioxidant balan­ce in the liver and blood plasma of rats with IR state.
In conclusion, the NSE administration to the rats with insulin-resistance state restored pro-/antioxidant balance and enhanced the content of nitric oxide, therefore, improving insulin sensitivity.

Signal function of cytokinin 6-benzylaminopurine in the reaction of Triticum aestivum L. mesophyll cells to hyperthermia

M. M. Musienko, V. V. Zhuk, L. M. Batsmanova

ESC Institute of Biology, Taras Shevchenko National University of Kyiv, Ukraine;
e-mail: zhuk_bas@voliacable.com

The signaling effect of 6-benzylaminopurine (BAP) on leaf mesophyll cells of Triticum aestivum L. under hyperthermic conditions was studied­. It was found that BAP regulated photosynthetic pigment, hydrogen peroxide content and activity of antioxidant enzymes, namely superoxide dismutase, ascorbate peroxidase and catalase under high-temperature conditions. The additive effect of BAP and high temperature on the activation of cell antioxidant systems was demonstrated. BAP regulated reducing processes in mesophyll leaf cells under high-temperature conditions.