Effect of hydroxytyrosol on the peroxidation of equine erythrocyte membranes by chemiluminescence
Keywords:
HT, equine erythrocyte membranes, Antioxidants, Peroxidation, ChemiluminescenceAbstract
Hydroxytyrosol (HT) is a polyphenol with a strong antioxidant effect, which is why it is being evaluated to improve animal and human health. The risk of injuries and performance of sporting horses are associated with their antioxidant status. Equine erythrocyte membranes are used as a peroxidation test model, due to their high content of double-linked fatty acids. Chemiluminescence is one of the most used methods to evaluate membrane peroxidation, due to its sensitivity and specificity. The objective of this work was to investigate the antioxidant effect of HT (POLYPHENOL - HT 1® Nova Mentis Ltd, Ireland) on the peroxidation of equine erythrocyte membranes. Erythrocyte membranes, also called ghosts, were adjusted to 1 mg of protein and challenged in a tert-butyl hydroperoxide (t-BHP)-dependent pro-oxidant model, at a 2 mM concentration and at 37 °C. Protection against peroxidation generated with HT (200 µM) was evaluated on it. Peroxidation of the ghosts was quantified in a Packard 1900 TR liquid scintillation counter using chemiluminescence in counts per minute (cpm). A completely randomized experimental design was used with the following groups: Control group (only erythrocyte membranes), 2mM t-BHP group (erythrocyte membranes + prooxidant substance) and HT group (erythrocyte membranes + prooxidant substance + HT200 µM). It was observed that the chemiluminescence value was statistically higher in the 2mM t-BHP group than in the Control, while the HT group showed an increasing reduction in chemiluminescence. The results of ten independent determinations per group show the mean and its standard deviation of the averages of 11224±2002 cpm (Control); 22000±3925 cpm (t-BHP 2mM) and 12784±2281 cpm (HT 200 μM). Data were statistically evaluated using one-way analysis of variance (ANOVA) and Tukey's test.In conclusion, HT protected equine erythrocyte membranes from peroxidation.
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Published on: 25-10-2024
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