Estimation of the Genotoxicityof Hydrogen Peroxide and Antioxidant Actionof Halo Acid by the Method of Dna-Cometwith the Use of the Model of Transplantable Cell Cultures

In the framework of the study, the degree of defragmentation of DNA by the DNA-comet method is evaluated when exposed to the cell culture of hydrogen peroxide (H2O2), and an in vitro model is developed to evaluate the antioxidant activity of new pharmacological agents. The results of working with cell lines show that the percentage of damage to the genetic material of cells of intact samples does not greatly vary from the method of removing the cellular monolayer from the culture plastic. Concerning the effect of H2O2 as an inducer of oxidative stress on DNA cell damage, the optimal level of DNA defragmentation has been modeled for subsequent studies of the protective action of antioxidants.

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Evaluation of low-grade In Vitro Oxidative Stress in Goat Erythrocytes ‎Exposed to Hydrogen Peroxide and Dexamethasone ‎

Sahel Journal of Veterinary Sciences ◽

10.54058/saheljvs.v18i2.244 ◽

2021 ◽

Vol 18 (2) ◽

pp. 33-37

Author(s):

M. B. Monguno ◽

E. S. Philip ◽

I. C. Uku ◽

I. O. Igbokwe

Keyword(s):

Oxidative Stress ◽

Hydrogen Peroxide ◽

In Vitro Model ◽

Sucrose Solution ◽

Osmotic Fragility ◽

Low Grade ◽

Mean Values ◽

Red Blood Cell Count ◽

Vitro Model

Under conditions of oxidative stress, erythrocytes of goats could be predisposed to haemolysis. This study was aimed at evaluating the effect of oxidant exposure to goat erythrocytes using an in vitro model. Blood samples from 10 goats were incubated with 0.06 – 0.18 mM hydrogen peroxide (H2O2) either singly or in combination with 0.02 µM dexamethasone for 60 min, and erythrocyte parameters such as packed cell volume (PCV), red blood cell count (RBC), mean corpuscular volume (MCV) and percentage haemolysis in hypotonic sucrose solution (250 mOsmol/L) were determined thereafter. No significant changes in the mean values of all parameters were observed. The in vitro model indicated that erythrocyte parameters remained stable under low-grade oxidant exposure in goats. Therefore, plasma H2O2 concentration of ≤ 0.18 mM, whether in the presence or absence of 0.02 µM dexamethasone, may not induce apparent oxidative damage in goat erythrocytes that could be estimated by PCV, RBC, MCV and sucrose-based osmotic fragility at low hypotonicity.

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Polyphenol-hydrogen peroxide reactions in skin: In vitro model relevant to study ROS reactions at inflammation

Analytica Chimica Acta ◽

10.1016/j.aca.2019.05.032 ◽

2019 ◽

Vol 1075 ◽

pp. 91-97 ◽

Cited By ~ 7

Author(s):

Mahboubeh Eskandari ◽

Jadwiga Rembiesa ◽

Lauryna Startaitė ◽

Anna Holefors ◽

Audronė Valančiūtė ◽

...

Keyword(s):

Hydrogen Peroxide ◽

In Vitro Model ◽

Vitro Model

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Epicatechin and its in vivo metabolite, 3′-O-methyl epicatechin, protect human fibroblasts from oxidative-stress-induced cell death involving caspase-3 activation

Biochemical Journal ◽

10.1042/bj3540493 ◽

2001 ◽

Vol 354 (3) ◽

pp. 493-500 ◽

Cited By ~ 69

Author(s):

Jeremy P. E. SPENCER ◽

Hagen SCHROETER ◽

Gunter KUHNLE ◽

S. Kaila S. SRAI ◽

Rex M. TYRRELL ◽

...

Keyword(s):

Oxidative Stress ◽

Hydrogen Peroxide ◽

Cell Death ◽

Caspase 3 ◽

Cell Damage ◽

Protective Action ◽

Human Fibroblasts ◽

Membrane Damage ◽

Antioxidant Properties

There is considerable current interest in the cytoprotective effects of natural antioxidants against oxidative stress. In particular, epicatechin, a major member of the flavanol family of polyphenols with powerful antioxidant properties in vitro, has been investigated to determine its ability to attenuate oxidative-stress-induced cell damage and to understand the mechanism of its protective action. We have induced oxidative stress in cultured human fibroblasts using hydrogen peroxide and examined the cellular responses in the form of mitochondrial function, cell-membrane damage, annexin-V binding and caspase-3 activation. Since one of the major metabolites of epicatechin in vivo is 3′-O-methyl epicatechin, we have compared its protective effects with that of epicatechin. The results provide the first evidence that 3′-O-methyl epicatechin inhibits cell death induced by hydrogen peroxide and that the mechanism involves suppression of caspase-3 activity as a marker for apoptosis. Furthermore, the protection elicited by 3′-O-methyl epicatechin is not significantly different from that of epicatechin, suggesting that hydrogen-donating antioxidant activity is not the primary mechanism of protection.

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Abstract 355: Effect of Poloxamer 188 on Rat Brain Isolated Mitochondria After Exposure to Hydrogen Peroxide

Circulation ◽

10.1161/circ.140.suppl_2.355 ◽

2019 ◽

Vol 140 (Suppl_2) ◽

Author(s):

Johannes A Pille ◽

Michele M Salzman ◽

Anna A Sonju ◽

Felicia P Lotze ◽

Josephine E Hees ◽

...

Keyword(s):

Hydrogen Peroxide ◽

Oxygen Consumption ◽

Mitochondrial Function ◽

Complex I ◽

Room Temperature ◽

In Vitro Model ◽

Atp Synthesis ◽

Complex Ii ◽

Vitro Model

Introduction: In a pig model of myocardial infarction (MI), intracoronary delivered Poloxamer (P) 188 significantly reduces ischemia/reperfusion (IR) injury when given immediately upon reperfusion, with improved mitochondrial function as a predominant effect. As mitochondria are heavily damaged during IR, a direct effect of P188 on mitochondria may lead to better therapy options during reperfusion. To show not only a similar reduction of IR injury by P188 in the brain, but also a direct P188 effect on mitochondria, we established an in-vitro model of IR that consists of damaging isolated rat brain mitochondria with hydrogen peroxide (H 2 O 2 ), one component of ischemia, then applying P188, and analyzing mitochondrial function. Methods: Male Sprague-Dawley rat brains were removed, and the mitochondria isolated by differential centrifugation and Percoll gradients, then kept on ice to slow their bioenergetics prior to any experimental treatments. Mitochondria were exposed to 200 μM H 2 O 2 for 10 min at room temperature with slight agitation; controls received no H 2 O 2 . Samples were then diluted ½ with buffer ± P188 (250 μM after dilution) to simulate reperfusion and treatment, and kept at room temperature for 10 further minutes. ATP synthesis was measured in a luminometer using a luciferase enzymatic assay. Oxygen consumption was measured by closed cell respirometry with an oxygen meter. In both assays, Complex I and Complex II were examined; Complex I substrates glutamate and malate, Complex II substrate succinate plus the Complex I inhibitor rotenone. Statistics: Data are expressed as mean ± SEM. One-Way ANOVA, SNK-Test; Kruskal-Wallis-Test; α=0.05, * vs control. Results: In both Complex I and II, mitochondrial function was significantly impaired by H 2 O 2 , with ATP synthesis affected more at Complex I and oxygen consumption affected more at Complex II. Addition of P188 did not provide any significant improvement in mitochondrial function. Conclusions: Although P188 significantly reduced IR injury when given during reperfusion in a pig model of MI, it does not appear to provide direct protection to mitochondria in this in-vitro model. Whether the exposure to H 2 O 2 causes the appropriate injury for P188 to become effective remains to be elucidated.

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2-Hydroxy-(4-methylseleno)butanoic Acid Is Used by Intestinal Caco-2 Cells as a Source of Selenium and Protects against Oxidative Stress

Journal of Nutrition ◽

10.1093/jn/nxz190 ◽

2019 ◽

Vol 149 (12) ◽

pp. 2191-2198

Author(s):

Joan Campo-Sabariz ◽

David Moral-Anter ◽

M Teresa Brufau ◽

Mickael Briens ◽

Eric Pinloche ◽

...

Keyword(s):

Oxidative Stress ◽

In Vitro Model ◽

Thioredoxin Reductase ◽

Protein Carbonyl ◽

Butanoic Acid ◽

H2o2 Treatment ◽

Vitro Model ◽

Gpx Activity

ABSTRACT Background Selenium (Se) participates in different functions in humans and other animals through its incorporation into selenoproteins as selenocysteine. Inadequate dietary Se is considered a risk factor for several chronic diseases associated with oxidative stress. Objective The role of 2-hydroxy-(4-methylseleno)butanoic acid (HMSeBA), an organic form of Se used in animal nutrition, in supporting selenoprotein synthesis and protecting against oxidative stress was investigated in an in vitro model of intestinal Caco-2 cells. Methods Glutathione peroxidase (GPX) and thioredoxin reductase (TXNRD) activities, selenoprotein P1 protein (SELENOP) and gene (SELENOP) expression, and GPX1 and GPX2 gene expression were studied in Se-deprived (FBS removal) and further HMSeBA-supplemented (0.1–625 μM, 72 h) cultures. The effect of HMSeBA supplementation (12.5 and 625 μM, 24 h) on oxidative stress induced by H2O2 (1 mM) was evaluated by the production of reactive oxygen species (ROS), 4-hydroxy-2-nonenal (4-HNE) adducts, and protein carbonyl residues compared with a sodium selenite control (SS, 5 μM). Results Se deprivation induced a reduction (P < 0.05) in GPX activity (62%), GPX1 expression, and both SELENOP (33%) and SELENOP expression. In contrast, an increase (P < 0.05) in GPX2 expression and no effect in TXNRD activity (P = 0.09) were observed. HMSeBA supplementation increased (P < 0.05) GPX activity (12.5–625 μM, 1.68–1.82-fold) and SELENOP protein expression (250 and 625 μM, 1.87- and 2.04-fold). Moreover, HMSeBA supplementation increased (P < 0.05) GPX1 (12.5 and 625 μM), GPX2 (625 μM), and SELENOP (12.5 and 625 μM) expression. HMSeBA (625 μM) was capable of decreasing (P < 0.05) ROS (32%), 4-HNE adduct (49%), and protein carbonyl residue (75%) production after H2O2 treatment. Conclusion Caco-2 cells can use HMSeBA as an Se source for selenoprotein synthesis, resulting in protection against oxidative stress.

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An Insight into Sargassum muticum Cytoprotective Mechanisms against Oxidative Stress on a Human Cell In Vitro Model

Marine Drugs ◽

10.3390/md15110353 ◽

2017 ◽

Vol 15 (11) ◽

pp. 353 ◽

Cited By ~ 3

Author(s):

Susete Pinteus ◽

Marco Lemos ◽

Joana Silva ◽

Celso Alves ◽

Agnieszka Neugebauer ◽

...

Keyword(s):

Oxidative Stress ◽

Human Cell ◽

In Vitro Model ◽

Sargassum Muticum ◽

Vitro Model ◽

Insight Into

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Trolox and Ascorbic Acid Reduce Direct and Indirect Oxidative Stress in the IPEC-J2 Cells, an In Vitro Model for the Porcine Gastrointestinal Tract

PLoS ONE ◽

10.1371/journal.pone.0120485 ◽

2015 ◽

Vol 10 (3) ◽

pp. e0120485 ◽

Cited By ~ 31

Author(s):

Hans Vergauwen ◽

Bart Tambuyzer ◽

Karen Jennes ◽

Jeroen Degroote ◽

Wei Wang ◽

...

Keyword(s):

Oxidative Stress ◽

Ascorbic Acid ◽

Gastrointestinal Tract ◽

In Vitro Model ◽

Vitro Model

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An Advanced In Vitro Model to Study Hypoxia/Low Glucose-Induced Neuronal Cell Damage and Death

Annals of the New York Academy of Sciences ◽

10.1111/j.1749-6632.1997.tb48437.x ◽

1997 ◽

Vol 825 (1 Neuroprotecti) ◽

pp. 267-278 ◽

Cited By ~ 4

Author(s):

M. P. TABOR ◽

H. B. WORP ◽

P. SODAAR ◽

H. VELDMAN ◽

E. A. J. JOOSTEN ◽

...

Keyword(s):

In Vitro Model ◽

Cell Damage ◽

Neuronal Cell ◽

Low Glucose ◽

Vitro Model

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Protective Effects and Mechanisms of Recombinant Human Glutathione Peroxidase 4 on Isoproterenol-Induced Myocardial Ischemia Injury

Oxidative Medicine and Cellular Longevity ◽

10.1155/2021/6632813 ◽

2021 ◽

Vol 2021 ◽

pp. 1-17

Author(s):

Chang Liu ◽

Bozhao Li ◽

Qi Yan ◽

Shaopeng Niu ◽

Yiding Zhao ◽

...

Keyword(s):

Oxidative Stress ◽

Myocardial Ischemia ◽

Glutathione Peroxidase ◽

In Vitro Model ◽

Cardiomyocyte Apoptosis ◽

Protective Effects ◽

Stress Injury ◽

Oxidative Stress Injury ◽

Vitro Model

Ischemic heart disease (IHD) is a cardiovascular disease with high fatality rate, and its pathogenesis is closely related to oxidative stress. Reactive oxygen species (ROS) in oxidative stress can lead to myocardial ischemia (MI) injury in many ways. Therefore, the application of antioxidants may be an effective way to prevent IHD. In recent years, glutathione peroxidase 4 (GPx4) has received increasing attention due to its antioxidant effect. In a previous study, we used the new chimeric tRNAUTuT6 to express highly active recombinant human GPx4 (rhGPx4) in amber-less Escherichia coli. In this study, we established an isoproterenol- (ISO-) induced MI injury model in rats and an in vitro model to research the protective effect and mechanism of rhGPx4 on MI injury. The results showed that rhGPx4 could reduce the area of myocardial infarction and ameliorate the pathological injury of heart tissue, significantly reduce ISO-induced abnormalities on electrocardiogram (ECG) and cardiac serum biomarkers, protect mitochondrial function, and attenuate cardiac oxidative stress injury. In an in vitro model, the results also confirmed that rhGPx4 could inhibit ISO-induced oxidative stress injury and cardiomyocyte apoptosis. The mechanism of action of rhGPx4 involves not only the inhibition of lipid peroxidation by eliminating ROS but also keeping a normal level of endogenous antioxidant enzymes by eliminating ROS, thereby preventing oxidative stress injury in cardiomyocytes. Additionally, rhGPx4 could inhibit cardiomyocyte apoptosis through a mitochondria-dependent pathway. In short, rhGPx4, a recombinant antioxidant enzyme, can play an important role in the prevention of IHD and may have great potential for application.

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