Tag Archives: oxidative stress

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.

Oxidative stress regulation in the yeast Ogataea polymorpha producer of human α-synuclein

N. V. Hrushanyk1, O. V. Stasyk2, O. G. Stasyk1*

1Ivan Franko National University of Lviv, Ukraine;
2Institute of Cell Biology, National Academy of Sciences of Ukraine, Lviv;
*e-mail: olenastasyk@gmail.com

Received: 02 March 2020; Accepted: 25 June 2020

In this study we analyzed how exogenous glucose levels affect enzymatic and non-enzymatic antioxidant defense systems and markers of oxidative stress in cells of the methylotrophic yeast Ogataea polymorpha producing recombinant human α-synuclein, implicated in pathogenesis of neurodegenerative Parkinson’s disease (PD). We found that glucose depletion up-induced activity of antioxidant enzymes superoxide dismutase, and catalase, and increased content of reduced and oxidized glutathione in the cells cultivated in the medium with 0.1% glucose, as compared to physiological growth condition (1% glucose-containing medium). In addition, low glucose concentration in the medium upregulated content of proteins carbonyl groups and of products of lipid peroxidation. Notably, the shift in the equilibrium toward pro-oxidant changes was similar for recombinant α-synuclein producer and parental wild-type strain. Thus, glucose limitation leads to the overproduction of reactive oxygen species in the methylotrophic yeast cells independently of the recombinant human α-synuclein production.

Effects of ethylthiosulfanylate and chromium (VI) on the state of pro/antioxidant system in rat liver

B. І. Kotyk1, R. Ya. Iskra1, O. M. Slivinska1, N. M. Liubas1,
A. Z. Pylypets1, V. I. Lubenets2, V. I. Pryimych3

1Institute of Animal Biology, NAAS of Ukraine, Lviv;
2Lviv Polytechnic National University, Ukraine;
3Stepan Gzhytskyi National University of Veterinary Medicine and Biotechnologies Lviv, Ukraine;
e-mail: kicyniabo@gmail.com

Received: 04 April 2020; Accepted: 25 June 2020

Ethylthiosulfanylate is alkyl ester of thiosulfoacid and belongs to the class of thiosulfonate compounds. Structurally, thiosulfonates are synthetic analogues of natural phytoncides. It is known that, natural organic sulfur-containing compounds are characterized by antioxidant and detoxification properties against heavy metals toxicity. Therefore, the purpose of the study was to investigate the influence of ethylthiosulfanylate, as a synthetic analogue of natural phytoncides, on the state of the pro/antioxidant system in the liver of laboratory rats exposed to Cr(VI). It was found that ethylthiosulfanylate exposure at a dose 100 mg/kg body weight daily for 14 days led to a decrease in the intensity of increasing of the lipid hydroperoxides (LHP) content in the rat liver caused by Cr(VI) action. In addition, ethylthiosulfanylate pretreatment prevented depletion of reduced glutathione (GSH) pool under the action of potassium dichromate oxidative stress and performed the accumulation of cellular GSH in rat liver.

Exogenous hydrogen sulfide for the treatment of mesenteric damage associated with fructose-induced malfunctions via inhibition of oxidative stress

O. Revenko1*, N. Zaichko2, J. Wallace3, O. Zayachkivska1

1Department of Physiology, Danylo Halytskyy Lviv National Medical University, Ukraine;
2Department of Biochemistry and General Chemistry,
National Pirogov Memorial Medical University, Vinnytsia, Ukraine;
3Department of Physiology and Pharmacology, University of Calgary, Canada;
*e-mail: wersus35@gmail.com

Received: 30 December 2019; Accepted: 27 March 2020

Remodeling of adipocytes in mesentery (AM) associated with nutritional overload from high fructose diet (HFD) is a source of several comorbidities. However, its pathogenesis is still unclear and there are no specific effective drugs for AM remodeling. Recently hydrogen sulfide (H2S) demonstrated potent cytoprotective actions. The purpose of this study was to investigate the effects and underlying mechanisms of AM remodeling in rats fed HFD and with H2S pre-treatment. Adult male rats on standard diet (SD, control group) or HFD that underwent acute water-immersion restraint stress (WIS) were evaluated for subcellular AM adaptive responses by electron microscopy. The effects on AM of exogenous sodium hydrosulfide (NaHS, 5.6 mg/kg/day for 9 days) and the Н2S-releasing aspirin (ASA) derivative (H2S-ASA [ATB-340], 17.5 mg/kg/day) vs conventional ASA (10 mg/kg/day) vs vehicle were investigated. Serum glucose level, thiobarbituric acid reactive substances (TBARS), and activities of cystathionine γ-lyase (CSE) and cystathionine β-synthase (CBS) were examined biochemically using spectrophotometry. In the HFD groups, treatment with NaHS protected AM, as mesenteric microvascular endothelial and sub-endothelial structures were observed vs the vehicle-treated group that had signs of endothelial dysfunction, AM damage and dysfunctional mitochondria. The effect of H2S-ASA was characterized by protection of AM against HFD and WIS-induced injury, with lower TBARS blood level and increased CSE and CBS activities. Carbohydrate overload for 4 weeks is sufficient to cause AM oxidative damage, mitochondrial dysfunction and endothelial changes. H2S plays an important role in mesenteric adipocyte cellular survival against HFD-induced oxidative stress by decreasing overproduction of TBARS and mitochondrial dysfunction. The use of H2S could lead to a novel approach for anti-obesity treatment.

Profiling of metabolic biomarkers in the serum of prostate cancer patients

F. Ali1, S. Akram1, S. Niaz1,2, N. Wajid1

1Institute of Molecular Biology and Biotechnology (IMBB) & Centre for Research In Molecular Medicine (CRIMM), The University of Lahore, Raiwind Road Lahore, Pakistan;
2Social Security Hospital Multan Chungi, Multan Road, Lahore;
e-mail: Fatima.ali@imbb.uol.edu.pk; fatemei.ali@gmail.com

Received: 26 July 2019; Accepted: 29 November 2019

Prostate cancer (PCa) is the major cause of the death of men population globally. Multiple factors are involved in the initiation and progression of PCa. This study aimed to evaluate different metabolic parameters in the serum of PCa patients. Males of 50 years and above age with the recent diagnosis of PCa (digital rectal examination, and elevated serum prostate-specific antigen (PSA) level) were included in the study. Glucose and serum electrolytes level, lactate dehydrogenase activity, parameters of lipid metabolism and liver and kidney functioning were measured on a fully automated analyzer using standard reagent kits. Oxidative stress was evaluated by measuring MDA, CAT, GSH, and SOD in serum. Detection of C-reactive protein (CRP), insulin-like growth factor (IGF-1) and vascular endothelial growth factor (VEGF) was performed by immunoassay. It was shown that serum glucose and HDL levels were lower while total cholesterol, LDL and triglyceride levels were significantly higher in PCa group than in the control group. PCa patients had an elevated level of liver and kidney functional markers. Comparison of the oxidative stress markers in patient and control groups showed significant difference. It was detected that serum levels of CRP, IGF-1 and VEGF were significantly higher in PCa group, compared the control to group (P < 0.05). Low level of glucose and dyslipidemia indices in prostate cancer patients indicated metabolic changes and demonstrated the importance of multiple parameters analysis (free PSA, dyslipidemia, VEGF, IGF-1, CRP, and oxidative stress markers) for early PCa diagnostics.

Oxidative stress and thiols depletion impair tibia fracture healing in young men with type 2 diabetes

H. I. Falfushynska1, O. I. Horyn1, D. V. Poznansky1, D. V. Osadchuk2,
T. О. Savchyn3, T. І. Krytskyi2, L. S. Merva1, S. Z. Hrabra1

1Ternopil Volodymyr Hnatiuk National Pedagogical University, Ukraine;
2I. Horbachevsky Ternopil National Medical University, Ukraine;
3Ternopil Ivan Puluj National Technical University, Ukraine;
e-mail: falfushynska@tnpu.edu.ua

Received: 05 May 2019; Accepted: 18 October 2019

Diabetes mellitus is a metabolic disorder that enhances fracture risk and hinders bone formation. The aim of the present study was to evaluate the parameters of oxidative stress, metallothioneins (MTs), metabolic changes and cytotoxicity signs in blood of young men with (DTF group) and without (TF group) type 2 diabetes (T2D) mellitus who had a tibia fracture due to trauma in relation to specific markers of bone formation. The level of reactive oxygen species was determined using a ROS-sensitive fluorescent dye dihydrorhodamine, DNA fragmentation was detected with Hoescht 33342 fluorescent dye and caspase-3 was assessed in terms of acetyl-Asp-Glu-Val-Asp p-nitroanilide. All other studied indices were determined by standard spectrometric methods. Our results revealed the significant effect of T2D on the bone healing. Indeed, the indices variation in the DTF group were significantly deeper as compared to group TF. The bone fracture in both TF and DTF groups had led to a significant decrease in antioxidants activity and/or level and a consistent increase in signs of oxidative damage. The concentration of MTs was also altered by trauma, but ina group-specific manner: an increase was noted in TF patients after trauma while in diabetes group a decrease in MTs was observed. Likewise, glutathione was strongly suppressed (by -64%) in DTF group. Tibia fracture provoked cytotoxicity which was manifested by increasing lactate dehydrogenase (LDH), cholinesterase and caspase-3 activity, the key effector of apoptosis in osteoclasts. The activity of alkaline phosphatase and total calcium increased only in TF group which demonstrated adequate remodelling process. The most prominent indices for groups splitting include ROS concentration, caspase 3, glutathione transferase and LDH activities mostly conjoint to DTF group. In sum, T2D impairs bone healing under condition of severe oxidative stress and cellular thiols depletion which result in an increase in apoptosis and DNA fragmentation. Our findings establish a biochemical link between increased oxidative stress and reduced bone markers and provide a rational for further studies investigating the role of pro- and antioxidants in bone healing.

The effect of quercetin on oxidative stress markers and mitochondrial permeability transition in the heart of rats with type 2 diabetes

N. I. Gorbenko1, O. Yu. Borikov2, O. V. Ivanova1, E. V. Taran1,
Т. S. Litvinova1, T. V. Kiprych1, A. S. Shalamai3

1V. Danilevsky Institute of Endocrine Pathology Problems, National Academy of Medical Sciences of Ukraine, Kharkiv;
2V. N. Karazin Kharkiv National University, Ukraine;
3PJSC SIC “Borshchahivskiy Chemical-Pharmaceutical Plant”, Kyiv, Ukraine;
е-mail: Gorbenkonat58@ukr.net

Received: 24 June 2019; Accepted: 13 August 2019

Increasing evidence suggests that oxidative stress and induction of mitochondrial permeability transition in cardiomyocytes are linked to tissue damage and the development of diabetic cardiovascular complications. The aim of this study was to assess the effects of quercetin (Q) on oxidative stress and mitochondrial permeability transition in the heart of rats with type 2 diabetes mellitus (DM). Type 2 DM was induced in 12-week-old male Wistar rats by intraperitoneal injections of 25 mg/kg streptozotocin twice per week followed by a high-fat diet during four weeks. The rats were divided into three groups: control intact group (C, n = 8), untreated diabetic group (Diabetes, n = 8) and diabetic rats treated with Q (50 mg/kg/day per os for 8 weeks) after diabetes induction (Diabetes+Q, n = 8). Administration of Q increased insulin sensitivity and normali­zed the functional state of cardiac mitochondria due to increased aconitase and succinate dehydrogenase activities in rats with type 2 DM. Q also ameliorated oxidative stress, decreasing the level of advanced oxidation protein products and increasing the activity of thioredoxin-reductase in heart mitochondria of diabetic rats. In addition, Ca2+-induced opening of the mitochondrial permeability transition pore was significantly inhibited in diabetic rats treated with Q in comparison with the untreated diabetic group. These data demonstrate that Q can protect against oxidative stress, mitochondrial permeability transition induction and mitochondrial dysfunction in cardiomyocytes of diabetic rats. We suggest that the use of Q may contribute to the amelioration of cardiovascular risk in type 2 DM.

Agonists of CB1 and NMDA receptors decrease the toxic effect of organophosphorus compound paraoxon on PC12 cells

F. Salem1, F. Bahrami1,2, Z. Bahari2, Z. Jangravi3, S. Najafizadeh-Sari4

1Neuroscience Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran;
2Department of Physiology and Medical Physics, Faculty of Medicine, Baqiyatallah University of Medical Sciences, Tehran, Iran;
3Departmentof Biochemistry, Faculty of Medicine, Baqiyatallah University
of Medical Sciences, Tehran, Iran;
4Student’ Research Committee (SRC), Baqiyatallah University of Medical Sciences, Tehran, Iran;
e-mail: f.bahrami@bmsu.ac.ir or farideh_bahrami@yahoo.com

Received: 01 July 2019; Accepted: 13 August 2019

Pharmacological studies allow to suggest that activation of cannabinoid type 1 receptors (CB1) have a neuroprotective role against toxicity induced by organophosphate agents, but the exact mechanisms of this effect as well as interaction with receptors of other types are far from clear. Therefore, the aim of current study was to evaluate the effect of CB1 and NMDA receptors agonists on cell viability and biomarkers of oxidative stress and lipid peroxidation in PC12 cells exposed to paraoxon. PC12 cells were exposed to 100 µm paraoxon as organophosphate agent. Treatments with 1 µM arachidonyl-2′-chloroethylamide (ACEA) as specific agonist of CB1 receptors, 100 µM N-methyl-D-aspartate (NMDA) as agonist of NMDA receptors and 1 µM AM251 as antagonist of CB1 receptors were done. Cell via­bility and biomarkers of oxidative stress were evaluated after 48 h of incubation. The level of CB1 receptor protein was evaluated by Western blotting. It was demonstrated that PC12 cells treatment with paraoxon led to cell viability inhibition, glutathione level, superoxide dismutase and catalase activity reduction, lipid peroxidation intensification and CB1 receptor expression attenuation. Application of ACEA and NMDA was shown to be followed by normalization of these indices. The protective effect of ACEA was abolished when the CB1 receptors antagonist AM251 was applied. The study revealed that application of ACEA and NMDA can protect PC12 cells against paraoxon induced toxicity through antioxidant capacity increment, lipid peroxidation inhibition and enhanced expression of CB1 receptors.

Carassius auratus as a novel model for the hyperglycemia study

H. I. Falfushynska1, O. I. Horyn1, L. L. Gnatyshyna1,2,
B. B. Buyak1, N. I. Rusnak1, O. O. Fedoruk1, O. B. Stoliar1

1Ternopil Volodymyr Hnatiuk National Pedagogical University, Ukraine;
e-mail: falfushynska@tnpu.edu.ua;
2I. Horbachevsky Ternopil State Medical University, Ukraine

Received: 21 August 2018; Accepted: 13 December 2018

The aim of the present study was to create a suitable model of the glucose toxicity and elucidate the ability of zinc-binding proteins metallothioneins in the crucian carp Carassius auratus to reflect it. For that, fish was loaded by three waterborne concentrations of glucose (low (5.55 mM, LC), middle (55.5 mM, MC) or high (111 mM, HC)) for 21 days. The level of blood glucose, responses of metallothioneins, oxidative stress, DNA instability in the liver, as well as erythrocytes indices, cholinesterase activity in the brain and morphometric variables were evaluated. An increase in blood glucose levels (up to 3–5 times), glycated hemoglobin (HbA1c, only by the HC, by 55%), methemoglobin (by two times), oxyradicals (16-57%) and TBARS levels (up to 57%), frequency of the micronucleated erythrocytes, DNA fragmentation in hepatocytes, body mass and hepatosomatic indices and a decrease in metallothioneins concentration (40-74%), cholinesterase activity (~70%), total hemoglobin (by 18%) and red blood cells count (only after HC-treatment, by 47%) were detected. The lysosomal membrane stability, evaluated by the neutral red retention time, was affected by all studied concentrations of glucose (decreased by 58%). The most prominent changes were observed after the HC of glucose. CART analysis revealed the significant splitting parameters for studied group differentiation including HbA1c, lysosomal membrane stability and lipid peroxidation. We could consider the crucian carp is a useful model organism to perform DM studies and in the future, this fish model can help in mechanistic investigations and testing therapeutic interventions under glycemic states.

Effects of alpha-ketoglutarate on lifespan and functional aging of Drosophila melanogaster flies

M. P. Lylyk1, M. M. Bayliak1, H. V. Shmihel1,
J. M. Storey2, K. B. Storey2, V. I. Lushchak1

1Vasyl Stefanyk Precarpathian National University, Ivano-Frankivsk, Ukraine;
2Institute of Biochemistry, Carleton University, Ottawa, Canada;
e-mail: lushchak@pu.if.ua; bayliak@ukr.net

The effects of an alpha-ketoglutarate-supplemented diet on lifespan and functional senescence were evaluated in the Canton S strain of Drosophila melanogaster. The results suggest that effects of dietary alpha-ketoglutarate (AKG) are dose- and gender-dependent. In males, diets containing 1-10 mM AKG did not affect mean and maximum lifespans, except that an increased maximum lifespan observed at 10 mM AKG. Diet with 20 mM AKG shortened median lifespan and had no effect on maximum lifespan of males. In females, diets with low concentrations of AKG (1 and 5 mM) did not affect lifespan, whereas diets supplemented with 10 and 20 mM AKG increased both median and maximum lifespans. At a lifespan-prolonging concentration (10 mM), AKG decreased fecundity, increased cold resistance and had no effect on climbing activity or resistance to oxi­dative stress in flies of either gender at middle (24 days) and old (40 days) ages. Moreover, middle-aged AKG-fed females but not males were more resistant to heat stress that was accompanied by higher levels of HSP90 protein as compared with controls. Middle-aged flies on AKG-supplemented diets showed elevated oxidative stress and had higher total protein and triacylglycerol levels as compared with controls. Hence, anti-aging effects of AKG do not seem to be related to preventing oxidative stress development but involve metabolic rearrangement and synthesis of specific protective proteins, which aid to resist destructive processes with age.