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Assessment of Oxidative Stress Biomarkers in Rat Blood
Redox status assessments are time-consuming, require a large volume of samples and great reagent amounts, and are not adequately described for methodological reproducibility. Here, the objective was to standardize redox balance determination, based on previously described spectrophotometric tests in...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Bio-Protocol
202
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9993082/ https://www.ncbi.nlm.nih.gov/pubmed/36908641 http://dx.doi.org/10.21769/BioProtoc.4626 |
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author | Sinzato, Yuri K. Rodrigues, Thiago Cruz, Larissa L. Barco, Vinícius S. Souza, Maysa R. Volpato, Gustavo T. Damasceno, Débora C. |
author_facet | Sinzato, Yuri K. Rodrigues, Thiago Cruz, Larissa L. Barco, Vinícius S. Souza, Maysa R. Volpato, Gustavo T. Damasceno, Débora C. |
author_sort | Sinzato, Yuri K. |
collection | PubMed |
description | Redox status assessments are time-consuming, require a large volume of samples and great reagent amounts, and are not adequately described for methodological reproducibility. Here, the objective was to standardize redox balance determination, based on previously described spectrophotometric tests in pregnant rats, to improve precision, time dispensed, and the volume of samples and reagents, while maintaining accuracy and adequate cost benefits. This protocol summarizes oxidative stress markers, which focus on spectrophotometric tests for the assessment of thiobarbituric acid–reactive substances, reduced thiol groups, and hydrogen peroxide, as well as the antioxidant activity of superoxide dismutase, glutathione peroxidase, and catalase in washed erythrocyte and serum samples from full-term pregnant rats. For non-pregnant rats and other species, it is necessary to standardize these determinations, especially the sample volume. All measurements were normalized by the estimated protein concentrations in each sample. To establish optimum conditions for the reproducibility of the proposed methods, we describe all changes made in each assay’s steps based on the reference method reassessed for the new standardizations. Furthermore, the calculations of the concentrations or activities of each marker are presented. Thus, we demonstrate that the analysis of serum samples is easier and faster, but it is impossible to detect catalase activity. Furthermore, the proposed methods can be applied for redox balance determination, especially using smaller reagent amounts and lower sample volumes in lesser time without losing accuracy, as is required in obtaining samples during rat pregnancy. |
format | Online Article Text |
id | pubmed-9993082 |
institution | National Center for Biotechnology Information |
language | English |
publishDate |
202 |
publisher | Bio-Protocol |
record_format | MEDLINE/PubMed |
spelling | pubmed-99930822023-03-09 Assessment of Oxidative Stress Biomarkers in Rat Blood Sinzato, Yuri K. Rodrigues, Thiago Cruz, Larissa L. Barco, Vinícius S. Souza, Maysa R. Volpato, Gustavo T. Damasceno, Débora C. Bio Protoc Methods Article Redox status assessments are time-consuming, require a large volume of samples and great reagent amounts, and are not adequately described for methodological reproducibility. Here, the objective was to standardize redox balance determination, based on previously described spectrophotometric tests in pregnant rats, to improve precision, time dispensed, and the volume of samples and reagents, while maintaining accuracy and adequate cost benefits. This protocol summarizes oxidative stress markers, which focus on spectrophotometric tests for the assessment of thiobarbituric acid–reactive substances, reduced thiol groups, and hydrogen peroxide, as well as the antioxidant activity of superoxide dismutase, glutathione peroxidase, and catalase in washed erythrocyte and serum samples from full-term pregnant rats. For non-pregnant rats and other species, it is necessary to standardize these determinations, especially the sample volume. All measurements were normalized by the estimated protein concentrations in each sample. To establish optimum conditions for the reproducibility of the proposed methods, we describe all changes made in each assay’s steps based on the reference method reassessed for the new standardizations. Furthermore, the calculations of the concentrations or activities of each marker are presented. Thus, we demonstrate that the analysis of serum samples is easier and faster, but it is impossible to detect catalase activity. Furthermore, the proposed methods can be applied for redox balance determination, especially using smaller reagent amounts and lower sample volumes in lesser time without losing accuracy, as is required in obtaining samples during rat pregnancy. Bio-Protocol 2023 -03- 05 /pmc/articles/PMC9993082/ /pubmed/36908641 http://dx.doi.org/10.21769/BioProtoc.4626 Text en Copyright © 2023 The Authors; exclusive licensee Bio-protocol LLC. https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Methods Article Sinzato, Yuri K. Rodrigues, Thiago Cruz, Larissa L. Barco, Vinícius S. Souza, Maysa R. Volpato, Gustavo T. Damasceno, Débora C. Assessment of Oxidative Stress Biomarkers in Rat Blood |
title |
Assessment of Oxidative Stress Biomarkers in Rat Blood
|
title_full |
Assessment of Oxidative Stress Biomarkers in Rat Blood
|
title_fullStr |
Assessment of Oxidative Stress Biomarkers in Rat Blood
|
title_full_unstemmed |
Assessment of Oxidative Stress Biomarkers in Rat Blood
|
title_short |
Assessment of Oxidative Stress Biomarkers in Rat Blood
|
title_sort | assessment of oxidative stress biomarkers in rat blood |
topic | Methods Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9993082/ https://www.ncbi.nlm.nih.gov/pubmed/36908641 http://dx.doi.org/10.21769/BioProtoc.4626 |
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