Tag Archives: antioxidant enzymes

Distribution and polymorphism of enzymes involved in antioxidant protection and xenobiotics biotransformation in the mediterranean mussel Mytilus galloprovincialis

V. А. Toptikov, I. Yu. Chubyk*, S. V. Chebotar

Odesa National Mechnуkov University, Ukraine;
*e-mail: i.chubyk@onu.edu.ua

Received: 18 July 2022; Revised: 17 October 2022;
Accepted: 04 November 2022; Available on-line: 14 November 2022

The aim of the work was to determine the tissue distribution, activity and polymorphism of 13 enzymes involved in antioxidative protection and xenobiotics biotransformation in a five bivalve mussel organs (hepatopancreas, ctenidia, mantle, leg, adductor muscles). Iso-allozyme analysis was performed by electrophoresis, correlation relationships between the studied enzymes in the whole mussel body were carried out using Spearman’s correlation analysis. It was found that all studied enzymes are polymorphic, each organ differed in the level of enzyme activity and a set of multiple forms. The coordinated functioning of protective enzymes in various mussel organs was demonstrated.

Protein content in the parental diet affects cold tolerance and antioxidant system state in the offspring Drosophila

O. M. Strilbytska1*, U. V. Semaniuk1, N. I. Burdyliuk1, O. V. Lushchak1,2*

1Department of Biochemistry and Biotechnology, Vasyl Stefanyk Precarpathian National University, Ivano-Frankivsk, Ukraine;
2Research and Development University, Ivano-Frankivsk, Ukraine;
*e-mail: olya_b08@ukr.net or oleh.lushchak@pnu.edu.ua

Received: 02 February 2021; Accepted: 21 January 2022

Dietary nutrients are the key determinants of the lifespan and metabolic health. The content of specific dietary compounds in the parental diet can epigenetically affect the physiological state of the offspring. Here, we studied how variable dietary protein content in the diet of parental generation affects antioxidant capacity of Drosophila melanogaster adult offspring. The dry yeast concentration ranging from 0.25% to 15% in the parental diet was the only variable in the experiments, whereas subsequent generation was kept on a diet of the same composition. We found, that flies fed with yeast-restricted (0.25%) diet produced F1 male flies with a higher cold tolerance and higher activity of the second-line antioxidant enzymes whereas in F1 females no effect of parental diet composition on the cold tolerance, catalase, GST, G6PDH, IDH activity and low thiols content was detected. The results suggest that nutrient-dependent changes of genes expression in the flies of paternal generation differently affect the stress response of males and females of the first-generation offspring.

Semicarbazide diminishes the signs of bleomycin-induced pulmonary fibrosis in rats

O. O. Hudkova*, I. P. Krysiuk, T. O. Kishko,
N. M. Popova, L. B. Drobot, N. V. Latyshko

Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
*e-mail: ogudkova@biochem.kiev.ua

Received: 16 July 2021; Accepted: 22 September  2021

The pathogenesis of pulmonary fibrosis (PF) is accompanied by extracellular matrix (ECM) deposition, oxidative stress, and inflammation progression, as well as hyperactivation of amine oxidases (AOs), which contribute to disease manifestation. The present study aims to elucidate the effect of semicarbazide (SC), an inhibitor of Cu-containing AOs: lysyl oxidase (LOX), semicarbazide sensitive amine oxidase (SSAO), diamine oxidase (DAO), on PF induced in rats by bleomycin (BLM). Eighteen male Wistar rats were randomly divided into four groups: Control, rats of BLM group received BLM (5 mg/kg, intratracheally once), BLM+SC group obtained 0.005% solution of SC (about 50 µg per capita per day) for three weeks starting immediately after BLM injection, and the Control+SC group drank the same solution as BLM+SC group. The content of cross-linked collagen in total bronchi and free radicals in lung, activities of LOX, SSAO, DAO, polyamine oxidase (PAO), Cu, Zn-superoxide dismutase (SOD1), catalase (CAT) and glutathione peroxidase (GPx) in lung and blood were measured. BLM injection induced PF that was confirmed histologically and morphometrically as well as by the elevation of the content of cross-linked collagen and free radicals. The activities of LOX and SSAO involved in post-translational modification of ECM and inflammation were significantly increased (P < 0.05). The activities of DAO, and PAO that control polyamine metabolism were also essentially raised. Among antioxidant enzymes, only GPx was activated in the BLM group as compared to control. These changes were absent in the BLM+SC group. SC intake promoted the fact that the histology and morphometric parameters of lung tissue, the content of cross-linked collagen in the bronchi and free radicals in the lung, as well as the activity of the studied enzymes remained at the control level. Our data suggest that SC suppresses the development of BLM-induced PF by inhibiting AOs activities.

Oxidative stress in the heart of rats exposed to acute intermittent hypobaric hypoxia

S. Dewi1*, M. Sadikin1, W. Mulyawan2

1Department of Biochemistry and Molecular Biology, Faculty of Medicine, Universitas Indonesia, Jakarta, Indonesia;
2Department of Aerophysiology, Lakespra Saryanto, Air Force Indonesian National Army, Jakarta, Indonesia;
*e-mail: syarifah.dewi@ui.ac.id

Received: 01 October 2020; Accepted: 17 May 2021

It is known that the altitude area causes hypoxic conditions due to the low oxygen partial pressure. This study was conducted to estimate oxidative stress indices in the heart tissue after Wister rats exposure to the acute intermittent hypobaric hypoxia. Hypobaric hypoxia exposure was simulated by keeping the rats in a hypobaric chamber for 1 min at 35,000 feet altitude. After that the altitude was gradually reduced to 30,000 and 25,000 feet and maitained for 5 min. 25 male Wistar rats were divided into control group and four treatment groups (I-IV), consisting of rats exposed 1, 2, 3 and 4 times to hypobaric hypoxia with a frequency once a week. The animals were removed from the experiment at the  height of 18,000 feet and the heart tissue was obtained. The carbonyl groups and  MDA levels and superoxide dismutase and  catalase activity were exami­ned in the supernatant of the heart tissue homogenate. In the samples of group I, the decrease  in catalase activity with a simultaneous notable increase in carbonyl groups level was observed compared to control. In the samples of groups III and IV, the carbonyl level normalized and the activity of  both antioxidant enzymes increased significantly. It was concluded that the increase of antioxidant enzymes activity can contribute to cardiac tissue adaptive response to acute hypobaric hypoxia exposure.

The effect of CO donor hemin on the antioxidant and osmoprotective systems state in Arabidopsis of a wild-type and mutants defective in jasmonate signaling under salt stress

M. A. Shkliarevskyi1, Yu. E. Kolupaev1,2*, T. O. Yastreb1,
Yu. V. Karpets1, A. P. Dmitriev3

1Dokuchaev Kharkiv National Agrarian University, Ukraine;
2Karazin Kharkiv National University, Ukraine;
3Institute of Cell Biology and Genetic Engineering, National Academy of Sciences of Ukraine, Kyiv;
*e-mail: plant.biology.knau@gmail.com

Received: 6 December 2020; Accepted: 17 May 2021

The role of the gasotransmitter carbon monoxide (CO) in signaling and adaptive processes in plants has been studied insufficiently. There are indirect data indicating jasmonate signaling participation in realization of CO effects, but  the possible connection between carbon monoxide and jasmonate signaling during plant adaptation to salt stress remains unclear. We studied the carbon monoxide donor hemin effect on the Arabidopsis of a wild-type (Col-0) and defective in jasmonate signaling coi1 and jin1 mutants response to the salt stress.  Arabidopsis thaliana 4-week-old plants were grown on a modified Hoagland’s medium. Plants were incubated for 24 h in usual or 2 µM hemin containing culture medium, then transferred to 150 mM NaCl containing media and incubated for 24 h before the medium was replaced with the usual one. It was shown that salt stress caused water deficiency and superoxide dismutase and catalase activity decrease in the plants of all three genotypes. Treatment with 2 μM hemin stabilized the levels of catalase activity and  photosynthetic pigments and increased guaiacol peroxidase activity in a wild-type, but not in  coi1 and jin1 mutant plants after  stress induction. Treated with hemin wild-type Arabidopsis plants accumulated more proline and sugars in response to stress than treated coi1 and jin1 mutants. It was concluded that jasmonate signaling can be involved in adaptive processes induced by exogenous carbon monoxide.

Effect of glutamic acid and cysteine on oxidative stress markers in rats

N. O. Salyha

Institute of Animal Biology, National Academy of Agrarian Sciences of Ukraine, Lviv;
е-mail: ynosyt@yahoo.com

Received: 7 May 2020; Accepted: 13 November 2020

Epinephrine (EPI) surges is known to be associated with stress induction and raising risk of heart strokes. The search for effective, nontoxic substances with antioxidative effects has been intensified in recent years. We focused our attention on two amino acids: L-glutamic acid (Glu) and L-cysteine (Cys). Our goal was to compare the effects of Glu, Cys and Glu in combination with Cys intraperitoneal administration on the antioxidant system indicators and the content of lipid peroxidation products in myocardium and spleen tissues of rats subjected to experimental EPI-induced stress. Rats were divided into five groups: EPI, EPI/Glu, EPI/Glu/Cys, EPI/Cys and control. The reduced glutathione (GSH) and TBA-active products level, glutathione peroxidase (GPx,), glutathione-S-transferase (GST), glutathione reductase (GR) and glucose-6-phosphate dehydrogenase (G6PDH) activity in tissues were measured. Our results indicate that epinephrine-induced stress increased the content of the lipid peroxidation products in myocardium and reduced the level of GSH in myocardium and spleen tissues of rats. Increasing of GPx activity in spleen only stressed animals were observed, while significantly lowered the GPx activity in groups of rats treated with amino acids (Glu, Glu/Cys, Cys). The obtained results suggest that the GR activity was significantly inhibited by stress in all investigated groups in spleen and epinephrine-induced rats and EPI/Cys groups of rats in myocardium. In rats treated with amino acids (particularly, Glu and Glu/Cys groups), we observed no significant difference in studied parameters. Our results indicate that application of Glu, Cys alone or in combination can increase GSH content in both studied tissues and activity of some antioxidative enzymes, and thus partially mitigated of epinephrine-induced stress in rats.

Adaptor protein Ruk/CIN85 affects redox balance in breast cancer cells

I. R. Horak*, N. V. Latyshko, O. O. Hudkova, T. O. Kishko,
O. V. Khudiakova, D. S. Gerashchenko, T. D. Skaterna,
I. P. Krysiuk, S. G. Shandrenko, L. B. Drobot

Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
*e-mail: iryna.horak@gmail.com

Received: 25 February 2020; Accepted: 15 May 2020

Excessive reactive oxygen species (ROS) production may lead to damage of cellular proteins, lipids and DNA, and cause cell death. Our previous findings demonstrated that increased level of adaptor protein Ruk/CIN85 contributes to breast cancer cells malignancy. The aim of this study was to investigate the role of Ruk/CIN85 in the maintaining of the redox balance in cancer cells. Mouse breast adenocarcinoma 4T1 cells with different levels of Ruk/CIN85 expression were used as a model in this study. Activities of catalase (CAT), glutathione peroxidase (GPx), superoxide dismutase (SOD), aldehyde dehydrogenase (ALDH) and formaldehyde dehydrogenase (FALDH), as well as H2O2 and aldehydes content were measured using fluorometric assays. Gene expression correlations between Ruk/CIN85 and antioxidant enzymes in breast cancer samples were analyzed using ist.medisapiens transcriptomic database. It was demonstrated that Ruk/CIN85-overexpressing 4T1 cells were characterized by increased production of H2O2 and reduced activities of CAT, GPx and SOD. Overexpression of Ruk/CIN85 resulted in decreased content of aldehydes together with increased activity of ALDH, while in Ruk/CIN85-knocked down 4T1 cells, activities of ALDH and FALDH were decreased. The data of transcriptomic analysis revealed the correlations between SH3KBP1 expression and CAT, GPX4, ALDH1A1, ALDH1L1, ALDH2, GSR, SOD1 in human breast carcinomas samples. The obtained results indicate that adaptor protein Ruk/CIN85 affects redox balance in mouse breast adenocarcinoma 4T1 cells.

Hydrogen peroxide as a signal mediator at inducing heat resistance in wheat seedlings by putrescine

Yu. E. Kolupaev1,2, A. I. Kokorev1, T. O. Yastreb1, E. I. Horielova1

1Dokuchaev Kharkiv National Agrarian University, Ukraine,
e-mail: plant_biology@ukr.net;
2Karazin Kharkiv National University, Ukraine

Received: 27 May 2019; Accepted: 18 October 2019

Polyamines are multifunctional stress metabolites of plants. However, information on the effect of exo­genous polyamines on plant resistance to high temperatures is contradictory, and it remains unclear which signal mediators are involved in the realization of their physiological effects. The possible involvement of hydrogen peroxide as a mediator under the action of exogenous diamine putrescine on the resistance of etiolated wheat seedlings (Triticum aestivum L.) to hyperthermia (10-minute heating at 46°C) and the functioning of antioxidant system was investigated. It was established that the treatment of seedlings with putrescine in 0.25–2.5 mM concentrations caused a significant increase in their heat resistance. In response to the putrescine effect, a transient increase in the H2O2 content occurred in the root cells. This effect was eliminated by treatment of seedlings with a diamine oxidase inhibitor aminoguanidine and an NADPH oxidase inhibitor imidazole. These inhibitors, as well as the scavenger of hydrogen peroxide dimethylthiourea (DMTU), mitigated the effects of increased heat resistance of seedlings and increased activity of superoxide dismutase and catalase caused by putrescine. Under the influence of DMTU and imidazole, but not aminoguanidine, the effect of increasing the activity of guaiacol peroxidase in the roots of seedlings treated with putrescine was eliminated. The conclusion was made about the role of hydrogen peroxide and the possible participation of diamine oxidase and NADPH oxidase in its formation during the implementation of the stress-protective effect of putrescine on wheat seedlings.

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.