Browsing by Author "Duarte, Filipa Maia"
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- Propolis Protects GC-1spg Spermatogonial Cells against Tert-Butyl Hydroperoxide-Induced Oxidative DamagePublication . Duarte, Filipa Maia; Feijó, Mariana; Luís, Ângelo; Socorro, Sílvia; Maia, Cláudio J.; Correia, SaraPropolis is a natural resin produced by honeybees with plenty of pharmacologic properties, including antioxidant activity. Oxidative stress disrupts germ cell development and sperm function, with demonstrated harmful effects on male reproduction. Several natural antioxidants have been shown to reduce oxidative damage and increase sperm fertility potential; however, little is known about the effects of propolis. This work evaluated the role of propolis in protecting spermatogonial cells from oxidative damage. Propolis’ phytochemical composition and antioxidant potential were determined, and mouse GC-1spg spermatogonial cells were treated with 0.1–500 µg/mL propolis (12–48 h) in the presence or absence of an oxidant stimulus (tert-butyl hydroperoxide, TBHP, 0.005–3.6 µg/mL, 12 h). Cytotoxicity was assessed by MTT assays and proliferation by Ki-67 immunocytochemistry. Apoptosis, reactive oxygen species (ROS), and antioxidant defenses were evaluated colorimetrically. Propolis presented high phenolic and flavonoid content and moderate antioxidant activity, increasing the viability of GC-1spg cells and counteracting TBHP’s effects on viability and proliferation. Additionally, propolis reduced ROS levels in GC-1spg, regardless of the presence of TBHP. Propolis decreased caspase-3 and increased glutathione peroxidase activity in TBHP-treated GC-1spg cells. The present study shows the protective action of propolis against oxidative damage in spermatogonia, opening the possibility of exploiting its benefits to male fertility.
- Protective potentialities of propolis towards spermatogonial oxidative damagePublication . Duarte, Filipa Maia; Correia, Sara Carina de Lima; Baptista, Cláudio Jorge Maia; Feijó, Mariana PombalMale 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.