Lycopene Offers Protection against Oxidative Damage in Frozen-Thawn Bovine Semen

Eva Tvrdá, Alica Mackovich, Hana Greifová, Norbert Lukáč


Excessive amounts of reactive oxygen species (ROS) associated with spermatozoa freezing and thawing cycles have the ability to cause oxidative stress, which may result in sperm membrane lipid peroxidation (LPO), DNA damage and apoptosis, leading to decreased sperm motility and fertilization ability. Lycopene, the most powerful singlet oxygen quencher of all carotenoids, may be a possible option for a more effective semen cryopreservation because of its antioxidant properties. By reacting with and neutralizing ROS, lycopene could reduce the incidence of oxidative stress and therefore, lessen the damage otherwise inflicted on frozen-thawn human or animal spermatozoa. This study focused to evaluate possible protective effects of lycopene on post-thawn bovine sperm and selected oxidative stress parameters. Ten bovine ejaculates were used in the study. Each ejaculate was splitted into two equal aliquots and diluted with a commercial semen extender containing lycopene (1.5 mM/L) or no supplement (control), cooled to 4°C, frozen and kept in liquid nitrogen. Frozen straws were thawn in a water bath for subsequent experiments. Computer assisted semen analysis was used to evaluate spermatozoa motility, ROS generation was quantified using luminol-based luminometry and LPO was assessed via the TBARS assay and UV/VIS spectrophotometry. Lycopene supplementation to the semen extender significantly (P<0.001) increased the post-thawn spermatozoa motility (50.80±1.43%) in comparison with the control (32.40±1.20%). Lycopene administration provided a significantly (P<0.001) higher protection against ROS overgeneration caused by semen freezing and thawing (17.80±2.82 RLU/sec/106 sperm) when compared to the lycopene-free control (36.19±3.41 RLU/sec/106 sperm). It was determined that the presence of lycopene in the semen extender resulted in a significantly (P<0.001) lower lipid peroxidation of the spermatozoa membranes (16.43±0.56 µM/L) in comparison to the control group (27.45±0.99 µM/L). In conclusion, lycopene improved bovine spermatozoa quality during the process of cryopreservation via its antioxidant capacity. Thus, lycopene supplementation may be recommended to facilitate the improvement of semen preservation in bovine breeding industry.


bulls; lipid peroxidation; lycopene; reactive oxygen species; spermatozoa

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