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      Antioxidant efficiency of lycopene on oxidative stress - induced damage in bovine spermatozoa

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          Abstract

          Background

          Lycopene (LYC) is a natural carotenoid with powerful reactive oxygen species (ROS) scavenging activities. The aim of this study was to investigate if lycopene has the ability to reverse ROS-mediated alterations to the motility, viability and intracellular antioxidant profile of bovine spermatozoa subjected to ferrous ascorbate (FeAA). Spermatozoa were washed out of fresh bovine semen, suspended in 2.9 % sodium citrate and subjected to LYC treatment (0.25, 0.5, 1 or 2 mmol/L) in the presence or absence of FeAA (150 μmol/L FeSO 4 and 750 μmol/L ascorbic acid) during a 6 h in vitro culture. Spermatozoa motion characteristics were assessed using the SpermVision™ computer-aided sperm analysis (CASA) system. Cell viability was examined with the metabolic activity (MTT) assay, ROS generation was quantified via luminometry 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 culture to investigate the intracellular activity of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) as well as the concentrations of glutathione (GSH) and malondialdehyde (MDA).

          Results

          FeAA treatment led to a reduced spermatozoa motility ( P < 0.001), viability ( P < 0.001) and a decline of the antioxidant capacity of spermatozoa ( P < 0.001) but increased the ROS generation ( P < 0.001), superoxide production ( P < 0.001) and lipid peroxidation ( P < 0.001). LYC administration resulted in a preservation of the spermatozoa motion parameters ( P < 0.001), mitochondrial activity ( P < 0.001) and antioxidant characteristics ( P < 0.001 with respect to SOD; P < 0.01 in relation to CAT; P < 0.05 as for GPx and GSH) with a concentration range of 1 and 2 mmol/L LYC revealed to be the most effective.

          Conclusions

          Our results suggest that LYC exhibits significant ROS-scavenging and antioxidant properties which may prevent spermatozoa alterations caused by oxidative stress, and preserve the functionality of male reproductive cells.

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          Most cited references38

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          Catalytic metals, ascorbate and free radicals: combinations to avoid.

          Trace levels of transition metals can participate in the metal-catalyzed Haber-Weiss reaction (superoxide-driven Fenton reaction) as well as catalyze the oxidation of ascorbate. Generally ascorbate is thought of as an excellent reducing agent; it is able to serve as a donor antioxidant in free radical-mediated oxidation processes. However, as a reducing agent it is also able to reduce redox-active metals such as copper and iron, thereby increasing the pro-oxidant chemistry of these metals. Thus ascorbate can serve as both a pro-oxidant and an antioxidant. In general, at low ascorbate concentrations, ascorbate is prone to be a pro-oxidant, and at high concentrations, it will tend to be an antioxidant. Hence there is a crossover effect. We propose that the "position" of this crossover effect is a function of the catalytic metal concentration. In this presentation, we discuss: (1) the role of catalytic metals in free radical-mediated oxidations; (2) ascorbate as both a pro-oxidant and an antioxidant; (3) catalytic metal catalysis of ascorbate oxidation; (4) use of ascorbate to determine adventitious catalytic metal concentrations; (5) use of ascorbate radical as a marker of oxidative stress; and (6) use of ascorbate and iron as free radical pro-oxidants in photodynamic therapy of cancer.
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            DNA damage to spermatozoa has impacts on fertilization and pregnancy.

            DNA damage in the male germ line has been associated with poor semen quality, low fertilization rates, impaired preimplantation development, increased abortion and an elevated incidence of disease in the offspring, including childhood cancer. The causes of this DNA damage are still uncertain but the major candidates are oxidative stress and aberrant apoptosis. The weight of evidence currently favours the former and, in keeping with this conclusion, positive results have been reported for antioxidant therapy both in vivo and in vitro. Resolving the causes of DNA damage in the male germ line will be essential if we are to prevent the generation of genetically damaged human embryos, particularly in the context of assisted conception therapy.
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              Apoptosis and DNA damage in human spermatozoa.

              DNA damage is frequently encountered in spermatozoa of subfertile males and is correlated with a range of adverse clinical outcomes including impaired fertilization, disrupted preimplantation embryonic development, increased rates of miscarriage and an enhanced risk of disease in the progeny. The etiology of DNA fragmentation in human spermatozoa is closely correlated with the appearance of oxidative base adducts and evidence of impaired spermiogenesis. We hypothesize that oxidative stress impedes spermiogenesis, resulting in the generation of spermatozoa with poorly remodelled chromatin. These defective cells have a tendency to default to an apoptotic pathway associated with motility loss, caspase activation, phosphatidylserine exteriorization and the activation of free radical generation by the mitochondria. The latter induces lipid peroxidation and oxidative DNA damage, which then leads to DNA fragmentation and cell death. The physical architecture of spermatozoa prevents any nucleases activated as a result of this apoptotic process from gaining access to the nuclear DNA and inducing its fragmentation. It is for this reason that a majority of the DNA damage encountered in human spermatozoa seems to be oxidative. Given the important role that oxidative stress seems to have in the etiology of DNA damage, there should be an important role for antioxidants in the treatment of this condition. If oxidative DNA damage in spermatozoa is providing a sensitive readout of systemic oxidative stress, the implications of these findings could stretch beyond our immediate goal of trying to minimize DNA damage in spermatozoa as a prelude to assisted conception therapy.
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                Author and article information

                Contributors
                evina.tvrda@gmail.com
                anton.kovacik@yahoo.com
                tusimova.eva@gmail.com
                dusanpaal@gmail.com
                a.mackovich@gmail.com
                jahongir.alimov@gmail.com
                norolukac@gmail.com
                Journal
                J Anim Sci Biotechnol
                J Anim Sci Biotechnol
                Journal of Animal Science and Biotechnology
                BioMed Central (London )
                1674-9782
                2049-1891
                6 September 2016
                6 September 2016
                2016
                : 7
                : 1
                : 50
                Affiliations
                [1 ]Department of Animal Physiology, Faculty of Biotechnology and Food Sciences, Slovak University of Agriculture in Nitra, Tr. A. Hlinku 2, Nitra, 94976 Slovakia
                [2 ]AgroBioTech Research Centre, Slovak University of Agriculture in Nitra, Tr. A. Hlinku 2, Nitra, 94976 Slovakia
                [3 ]Department of Botany and Genetics, Faculty of Natural Sciences, Constantine the Philosopher University in Nitra, Nábrežie mládeže 91, Nitra, 94974 Slovakia
                [4 ]Department of General Biology, Faculty of Natural Sciences, Gulistan State University, 4th Microrayon, Guliston, 120100 Syrdarya Uzbekistan
                Author information
                http://orcid.org/0000-0003-2895-1249
                Article
                113
                10.1186/s40104-016-0113-9
                5011861
                27602206
                7d53d871-b03c-460b-adb5-344ec6241d6e
                © The Author(s). 2016

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 24 March 2016
                : 24 August 2016
                Funding
                Funded by: FundRef http://dx.doi.org/http://dx.doi.org/10.13039/501100005357, Agentúra na Podporu Výskumu a Vývoja;
                Award ID: APVV-0304-12
                Funded by: FundRef http://dx.doi.org/10.13039/501100006109, Vedecká Grantová Agentúra MVVa SR a SAV;
                Award ID: 1/0857/14
                Funded by: FundRef http://dx.doi.org/http://dx.doi.org/10.13039/501100003194, Agentúra Ministerstva kolstva, vedy, výskumu a portu SR;
                Award ID: ITMS 26220220180
                Categories
                Research
                Custom metadata
                © The Author(s) 2016

                Animal science & Zoology
                antioxidants,bulls,ferrous ascorbate,lycopene,oxidative stress,spermatozoa
                Animal science & Zoology
                antioxidants, bulls, ferrous ascorbate, lycopene, oxidative stress, spermatozoa

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