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  • Protective potentialities of propolis towards spermatogonial oxidative damage
    Publication . Duarte, Filipa Maia; Correia, Sara Carina de Lima; Baptista, Cláudio Jorge Maia; Feijó, Mariana Pombal
    Male fertility is dependent on successful spermatogenesis, as well as an array of genetic, environmental, and physiological factors. In this context, oxidative damage in testicular tissue affects the reproductive system, leading to 30 to 80% of infertile cases. High levels of reactive oxygen species (ROS) have been reported to disrupt the development of germ cells and to interfere with sperm function. Several natural antioxidants have been described to prevent or reduce this damage. Propolis is a natural resinous mixture produced by honeybees with plenty of pharmacologic and biological properties, including antioxidant capacity. Hence, propolis is a good candidate to protect the male reproductive system against oxidative stress (OS). Moreover, despite all the evidence about the protective potential of propolis, there are no reports directly focused on the effect of propolis on specific testicular cell populations. Following this rationale, the present dissertation aimed to analyse the role of propolis in protecting spermatogonial cells from oxidative damage. After evaluating the phytochemical composition and the antioxidant activity of propolis, type B spermatogonia cells (GC-1spg) were treated with propolis (0.1-500 µg/mL; 12-48 hours), in the presence or the absence of an oxidant stimulus (tert-butyl hydroperoxide, TBHP, 0.005-3.6 µg/mL, 12 hours). The cytotoxicity of both propolis and TBHP mingled or not, was analysed by 3-(4,5-dimethylthiazol-2- yl)-2,5-diphenyltetrazolium bromide (MTT) assay and proliferation by Ki-67 immunocytochemistry. The apoptosis rate, ROS levels, and antioxidant defences were studied through colorimetric methods. The obtained results showed a high content of total phenolics and flavonoids in the propolis extract, and moderate antioxidant activity. Propolis (0.1 µg/mL) increased the viability of GC-1spg cells, counterbalancing the impact of TBHP exposure (1.8 µg/mL). Additionally, regardless of TBHP presence, propolis reduced ROS levels in GC-1spg cells. In the presence of TBHP, propolis decreased caspase-3 activity and attenuated the TBHP-induced decrease in proliferation rate, increasing glutathione peroxidase activity. The present work highlighted the benefits of natural compounds to male fertility, more precisely, the protective action of propolis against TBHP-induced OS in spermatogonial cells (GC-1spg). Indeed, the disruptive actions of TBHP were counteracted, or even neutralized in the presence of propolis. These findings highlight propolis role as a protective agent against OS, which is crucial in the context of spermatogenesis and male fertility.
    2024-02-15Master thesis Open access Show more