Abstract
Simple SummaryIn living organisms, the antioxidant defense system serves to counteract reactive oxygen (ROS) and nitrogen (RNS) species, thereby protecting cellular targets against their oxidative damage; it includes a combination of different substances of endogenous or exogenous origin. Several methods were developed to assess the overall antioxidant capacity or the precise determination of individual key antioxidants. In the present study, the total antioxidant capacity (TAC) in healthy and dexamethasone-stressed hen serum was measured by applying four different spectrophotometric methods intended for both clinical and research studies, which could be automated on clinical auto-analyzers, thus allowing rapid and not expensive data collections. TAC values assessed by all four methods did not change throughout the experimental period in the control group, whereas significant changes were shown by all adopted assays in the stressed group, with some remarkable differences, probably due to the different contribution in each assay of the various antioxidant substances present in the samples. Therefore, when TAC evaluation is necessary to verify if animals are experiencing oxidative stress (OS) or to evaluate possible benefits from an antioxidant-enriched diet, TAC assessment should involve multiple assays, due to the different analytical technologies on which their assessments are based.Total antioxidant capacity (TAC) in healthy and dexamethasone-stressed hens was measured by applying four different spectrophotometric methods—the ferric reducing ability (FRAP) assay, the 2,2′-azino-bis (3-ethylbenzotiazoline-6-sulphonic acid) (ABTS) radical cation decolorization assay, the free radical scavenging activity (FRSA), and the total thiol levels (TTL). TAC assessed by all four methods did not change throughout the experimental period in the control group, whereas significant changes were shown by all adopted assays in the stressed group with some remarkable differences. TAC increased in the stressed group when FRAP and ABTS assays were applied, while it was reduced when sera were assessed by FRSA and TTL assays. Furthermore, FRAP assay was the only test able to show a significant change in TAC immediately after the end of the induced stress. At the end of the experimental period, TAC assessed by ABTS and FRSA assays showed a complete recovery in the stressed group, whereas TAC assessed by FRAP and TTL assays still showed significant persistent differences when compared to the control group. The observed differences in TAC are discussed in the light of the different contribution in each assay of the various antioxidant substances present in the samples.
Highlights
In living organisms, the antioxidant defense system serves to counteract reactive oxygen (ROS)and nitrogen (RNS) species, thereby protecting cellular targets against their oxidative damage.Animals 2020, 10, 2019; doi:10.3390/ani10112019 www.mdpi.com/journal/animalsThe antioxidant system includes a combination of different substances, of endogenous or exogenous origin, which all act together to detoxify oxidative substances to either delay or prevent the oxidative damage of macromolecules [1].Several methods were developed to assess the overall antioxidant capacity or the precise determination of individual key antioxidants
One of the critical points adduced to the application of the assays measuring the cumulative antioxidant potential of biological samples is that the results obtained with different methods are not always comparable, often returning inconsistent data, depending on the different technology used for their assessment [4,5,6]
Two-way repeated measures analysis of variance (ANOVA) showed that neither total protein nor albumin concentrations were affected by the independent variables (p > 0.05, data not shown)
Summary
The antioxidant defense system serves to counteract reactive oxygen (ROS)and nitrogen (RNS) species, thereby protecting cellular targets against their oxidative damage.Animals 2020, 10, 2019; doi:10.3390/ani10112019 www.mdpi.com/journal/animalsThe antioxidant system includes a combination of different substances, of endogenous (e.g., antioxidant enzymes, GSH, etc.) or exogenous origin (externally supplied by diets, e.g., vitamin C and E, carotenoids, etc.), which all act together to detoxify oxidative substances to either delay or prevent the oxidative damage of macromolecules [1].Several methods were developed to assess the overall antioxidant capacity or the precise determination of individual key antioxidants. The methods for the evaluation of the total antioxidant capacity (TAC) are generally based on the content of free radicals scavenged by a test solution, or on the capacity to reduce an oxidized chemical substance [2,3]. One of the main advantages of these tests is that they return the activity of all antioxidants of the biological sample into a single value, providing an integrated parameter rather than the simple sum of measurable substances. Thereby, the obtained values are considered as the cumulative effect of all antioxidants of the biological samples. One of the critical points adduced to the application of the assays measuring the cumulative antioxidant potential of biological samples is that the results obtained with different methods are not always comparable, often returning inconsistent data, depending on the different technology used for their assessment [4,5,6]
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