Resveratrol offers protection to oxidative stress induced by ferrous ascorbate in bovine spermatozoa

Abstract

Resveratrol (RES) is a natural polyphenol and phytoestrogen exhibiting cardioprotective, anticancer, antibacterial and vasorelaxing properties. It is also a powerful reactive oxygen species (ROS) scavenger and chelating agent. This study was designed to determine the efficiency of RES to reverse the ROS-mediated impairment of the motility, viability and intracellular antioxidant profile of bovine spermatozoa. Spermatozoa were washed out of fresh bovine semen, suspended in 2.9% sodium citrate and subjected to RES treatment (5, 10, 25 and 50 μmol L⁻¹) in the presence or absence of a pro-oxidant, i.e., ferrous ascorbate (FeAA; 150 μmol L⁻¹ FeSO₄ and 750 μmol L⁻¹ ascorbic acid) during a 6-h in vitro culture. Spermatozoa motion parameters were assessed using the SpermVision computer-aided sperm analysis (CASA) system. Cell viability was examined with the metabolic activity (MTT) assay, and the nitroblue-tetrazolium (NBT) test was applied to quantify the intracellular superoxide formation. Cell lysates were prepared at the end of the in vitro experiments in order to investigate the intracellular activity of superoxide dismutase (SOD), catalase (CAT), as well as the concentrations of glutathione (GSH) and malondialdehyde (MDA). FeAA treatment led to a reduced sperm motility (P < 0.001) and viability (P < 0.001), decreased the antioxidant parameters of the samples (P < 0.001 in case of SOD; P < 0.01 with respect to CAT; P < 0.05 in relation to GSH) but increased the superoxide production (P < 0.001) and lipid peroxidation (P < 0.001). RES supplementation resulted in a preservation of the spermatozoa vitality and antioxidant characteristics (P < 0.001 in case of SOD; P < 0.01 with respect to 25–50 μmol L⁻¹ RES and P < 0.05 in relation to 10 μmol L⁻¹ RES; P < 0.05 in case of GSH), with 50 μmol L⁻¹ RES proving to be the most effective RES concentration. Our results suggest that RES possesses significant antioxidant properties that may prevent the deleterious effects caused by ROS to spermatozoa, and preserve the fertilization potential of male reproductive cells.

Keywords:

• Resveratrol
• spermatozoa
• oxidative stress
• ferrous ascorbate
• bulls
• antioxidants

Funding

This work was co-funded by the European Community under the Project no. 26220220180: Building Research Centre “AgroBioTech” and the Scientific Grant Agency of the Ministry of Education of the Slovak Republic and of the Slovak Academy of Sciences VEGA Project No. 1/0857/14.