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Effect of Trolox and resveratrol supplementation during the refrigeration of boar sperm
Efecto de la suplementación con Trolox y resveratrol durante la refrigeración de semen porcino
DOI:
https://doi.org/10.15446/rfnam.v78n2.114567Keywords:
Antioxidant , Oxidative stress, Spermatozoa, Sperm preservation, Sperm viability (en)Antioxidante, Estrés oxidativo, Espermatozoides, Conservación espermática, Viabilidad espermática (es)
This research aimed to evaluate the possibility of improving the boar semen quality already refrigerated in commercial diluent by adding Trolox and/or resveratrol during the refrigeration process. Pools of refrigerated semen from boars of proven fertility were added with a) 200 µM Trolox, b) 50 µM resveratrol, c) 200 µM Trolox + 50 µM resveratrol, or d) no antioxidant supplementation (negative control) and conserved at 17 °C for 7 days. On days 1, 3 and 7 of refrigeration the following sperm parameters were evaluated: Motility, functions of the plasma membrane, pre-capacitated spermatozoa, viability and acrosome integrity, and mitochondrial membrane potential. Additionally, mitochondrial superoxide anion production was evaluated by flow cytometry. In vitro fertilization was performed using semen from day 3 of refrigeration. Data were analyzed using ANOVA (P<0.05). Evaluated parameters significantly decreased over refrigeration time (P<0.05), except the percentages of pre-capacitated spermatozoa. Samples supplemented only with Trolox maintained values similar to the control group, except for higher sperm motility on day 3 of preservation (P<0.05), whereas the addition of resveratrol caused a decrease in the studied parameters (P<0.05); the combination of both antioxidants showed intermediate values between each individual antioxidant treatment. No significant variations in mitochondrial superoxide anion production and cleavage rates were detected with each treatment with respect to the control. In conclusion, supplementing boar semen with Trolox and/or resveratrol does not cause a significant improvement in semen quality, once the refrigeration process has started, except for a higher motility with Trolox at day 3 of refrigeration, nor does it alter the mitochondrial production of reactive oxygen species and the fecundity capacity of semen in vitro.
El objetivo del presente trabajo fue evaluar la posibilidad de mejorar la calidad de semen porcino ya refrigerado en diluyente comercial por el agregado de Trolox y/o resveratrol durante el proceso de refrigeración. Mezclas de semen refrigerado provenientes de verracos de probada fertilidad fueron adicionadas con a) Trolox 200 µM, b) resveratrol 50 µM, c) Trolox 200 µM + resveratrol 50 µM, od) sin suplementación antioxidante (control negativo) y conservados a 17 °C por 7 días. En los días 1, 3 y 7 de la refrigeración, se evaluaron los siguientes parámetros espermáticos: motilidad, funcionalidad de la membrana plasmática, espermatozoides precapacitados, viabilidad e integridad acrosomal y potencial de membrana mitocondrial. Además, se evaluó la producción de anión superóxido mitocondrial utilizando citometría de flujo. La fecundación in vitro se realizó usando semen del día 3 de refrigeración. Los datos fueron analizados por Análisis de Varianza (P<0,05). Los parámetros evaluados disminuyeron con el tiempo de refrigeración (P<0,05), excepto por los porcentajes de espermatozoides precapacitados. Las muestras suplementadas solo con Trolox mantuvieron valores similares a las del grupo control, excepto por una mayor motilidad espermática en el día 3 de conservación (P<0,05), mientras que la adición de resveratrol causó una disminución de los parámetros estudiados (P<0,05), la combinación de ambos antioxidantes mostró valores intermedios entre cada tratamiento antioxidante individual. No se observaron variaciones en los porcentajes de producción de anión superóxido mitocondrial y de clivaje embrionario en respuesta a la suplementación con cada antioxidante con respecto al control. En conclusión, la suplementación de semen porcino con Trolox y/o resveratrol, una vez que el proceso de refrigeración ha comenzado, no provoca un aumento significativo de la calidad seminal, excepto por una mayor motilidad con Trolox al día 3, ni altera la producción mitocondrial de especies reactivas del oxígeno y la capacidad fecundante del semen in vitro.
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