In vitro Effects of Salvia officinalis on Bovine Spermatozoa

Authors: Eva Tvrdá, Boris Botman, Marek Halenár, Tomáš Slanina, Norbert Lukáč

Abstract:

In vitro storage and processing of animal semen represents a risk factor to spermatozoa vitality, potentially leading to reduced fertility. A variety of substances isolated from natural sources may exhibit protective or antioxidant properties on the spermatozoon, thus extending the lifespan of stored ejaculates. This study compared the ability of different concentrations of the Salvia officinalis extract on the motility, mitochondrial activity, viability and reactive oxygen species (ROS) production by bovine spermatozoa during different time periods (0, 2, 6 and 24 h) of in vitro culture. Spermatozoa motility was assessed using the Computer-assisted sperm analysis (CASA) system. Cell viability was examined using the metabolic activity MTT assay, the eosin-nigrosin staining technique was used to evaluate the sperm viability and ROS generation was quantified using luminometry. The CASA analysis revealed that the motility in the experimental groups supplemented with 0.5-2 µg/mL Salvia extract was significantly lower in comparison with the control (P<0.05; Time 24 h). At the same time, a long-term exposure of spermatozoa to concentrations ranging between 0.05 µg/mL and 2 µg/mL had a negative impact on the mitochondrial metabolism (P<0.05; Time 24 h). The viability staining revealed that 0.001-1 µg/mL Salvia extract had no effects on bovine male gametes, however 2 µg/mL Salvia had a persisting negative effect on spermatozoa (P<0.05). Furthermore 0.05-2 µg/mL Salvia exhibited an immediate ROS-promoting effect on the sperm culture (P>0.05; Time 0 h and 2 h), which remained significant throughout the entire in vitro culture (P<0.05; Time 24 h). Our results point out to the necessity to examine specific effects the biomolecules present in Salvia officinalis may have individually or collectively on the in vitro sperm vitality and oxidative profile.

Keywords: Bulls, CASA, MTT test, reactive oxygen species, sage, Salvia officinalis, spermatozoa.

Digital Object Identifier (DOI): doi.org/10.5281/zenodo.1339844

References:

[1] F.M. Lanzafame, S. La Vignera, E. Vicari, and A.E. Calogero, “Oxidative stress and medical antioxidant treatment in male infertility,” Reproductive Biomedicine Online, vol. 19, pp. 638-659, 2009.
[2] E. Tvrdá, Z. Kňažická, L. Bárdos, P. Massányi, and N. Lukáč, “Impact of oxidative stress on male fertility - a review,” Acta Veterinaria Hungarica, vol. 59, pp. 465-484, 2011.
[3] A. K. Bansal, and G. S. Bilaspuri, “Impacts of oxidative stress and antioxidants on semen functions,” Veterinary Medicine International, 2011, doi:10.4061/2011/686137.
[4] P. Gharagozloo, and R.J. Aitken RJ, “The role of sperm oxidative stress in male infertility and the significance of oral antioxidant therapy,” Human Reproduction, vol. 26, pp. 1628-1640, 2011.
[5] K. Tremellen, “Oxidative stress and male infertility - a clinical perspective,” Human Reproduction Update, vol. 14, pp. 243-258, 2008.
[6] G.N. De Iuliis, J.K. Wingate, A.J. Koppers, E.A. McLaughlin, and R.J. Aitken, “Definitive evidence for the nonmitochondrial production of superoxide anion by human spermatozoa,” Journal of Clinical Endocrinology & Metabolism, vol 91, pp. 1968-1975, 2006.
[7] R.J. Aitken, M.A. Baker, G.N. De Iuliis, and B. Nixon, “New insights into sperm physiology and pathology,” Handbook of Experimental Pharmacology, vol. 1989, p. 99-115, 2010.
[8] A. Butler, X. He, R.E. Gordon, H.S. Wu, S. Gatt, and E.H. Schuchman, “Reproductive pathology and sperm physiology in acid sphingomyelinase-deficient mice,” American Journal of Pathology, vol. 161, pp. 1061-1075, 2002.
[9] S.A. Sakr, and G.M. Badawy, “Effect of ginger (Zingiber officinale R.) on met ram-inhibited spermatogenesis and induced apoptosis in albino mice,” Journal of Applied Pharmaceutical Science, vol. 1, pp. 131-136, 2011.
[10] S.D. Santos-Filho, A. da Fonseca, and M. Bernardo-Filho, “The male reproductive system and the effect of an extract of a medicinal plant (Hypericum perforatum) on the labeling process of blood constituents with technetium- 99m,” Brazilian Archives of Biology and Technology, vol. 50, pp. 97-104, 2007.
[11] S.J. Kakel, and S.M. Ahmed, S.M., “Effect of Thymus vulgaris oil on some reproductive characters in adult male rats,” Iraqi Journal of Veterinary Science, vol. 2, pp. 83-87, 2008.
[12] A.A. Tohamy, S.R. Ibrahim, and A.E. Abdel Moneim, A.E., “Studies on the effect of Salvia aegyptiaca and Trigonella foenum graecum extracts on adult male mice,” Journal of Applied Pharmaceutical Science, vol. 2, pp. 36-43, 2012.
[13] R. Putheti, and R.N. Okigbo, “Effects of plants and medicinal plant combinations as antiinfective,” African Journal of Pharmacy and Pharmacology, vol. 2, pp. 130-135, 2008.
[14] Z. Omogbadegun, C. Uwadia, C. Ayo, V. Mbarika, N. Omoregbe, E. Otofia, and F. Chieze, “Multimedia-based medicinal plants sustainability management system,” International Journal of Computer Science, vol. 8, pp. 1694-0814, 2011.
[15] M. Hamidpour, R. Hamidpour, S. Hamidpour, and M. Shahlari, “Chemistry, pharmacology, and medicinal property of sage (Salvia) to prevent and cure illnesses such as obesity, diabetes, depression, dementia, lupus, autism, heart disease, and cancer,” Journal of Traditional and Complementary Medicine, vol. 4, pp. 82-88, 2014.
[16] S. Mora, R. Millán, H. Lungenstrass, G. Díaz-Véliz, J.A. Morán, M. Herrera-Ruiz, and J. Tortoriello, “The hydroalcoholic extract of Salvia elegans induces anxiolytic- and antidepressant-like effects in rats,” Journal of Ethnopharmacology, vol. 15, pp. 76-81, 2006.
[17] E.Y. Qnais, M. Abu-Dieyeh, F.A. Abdulla, and S.S. Abdalla, “The antinociceptive and anti-inflammatory effects of Salvia officinalis leaf aqueous and butanol extracts,” Pharmaceutical Biology, vol. 48, pp. 1149-1156, 2010.
[18] M.H. Hamdy Roby, M.A. Sarhan, K. Abdel-Hamed Selim, and K. I. Khalel, “Evaluation of antioxidant activity, total phenols and phenolic compounds in thyme (Thymus vulgaris L.), sage (Salvia officinalis L.), and marjoram (Origanum majorana L.) extracts,” Industrial Crops and Products, vol. 43, pp. 827-831, 2013.
[19] C.G.A. van Veldhuizen-Staas, “Overabundant milk supply: an alternative way to intervene by full drainage and block feeding,” International Breastfeeding Journal, vol. 2, pp. 11, 2007.
[20] S. Rahte, R. Evans, P.J. Eugster, L. Marcourt, J.L. Wolfender, A. Kortenkamp, and D. Tasdemir, “Salvia officinalis for hot flushes: towards determination of mechanism of activity and active principles,” Planta Medica, vol. 79, pp. 753-760, 2013.
[21] B. Bozin, N. Mimica-Dukic, I. Samojlik, and E. Jovin, “Antimicrobial and antioxidant properties of rosemary and sage (Rosmarinus officinalis L. and Salvia officinalis L., Lamiaceae) essential oils,” Journal of Agricultural and Food Chemistry, vol. 55, pp. 7879-7885, 2007.
[22] M. Koşar, H.J. Dorman, K.H. Başer, and R. Hiltunen, “Salvia officinalis L.: composition and antioxidant-related activities of a crude extract and selected sub-fractions,” Natural Product Communications, vol. 5, pp. 1453-1456, 2010.
[23] H. Al-Ahmed, R. Mahood, G. Bonev, and S. Georgieva, “Effects of antioxidant (Salvia officinalis L. extract) on in vitro fertilization and micromanipulation intracytoplasmic sperm injection in Albino mice,” Agricultural Science and Technology, vol. 4, pp. 15-19, 2012.
[24] M.H. Al-Malaly, A.F. Al-Abedy, and F.M. Al-Zubaidy, “Effect of different level of Zingiber officinale and Salvia officinalis on semen diluents for Awassi ram,” Journal of Genetic and Environmental Resources Conservation, vol. 1, pp. 67-70, 2013.
[25] A. Montón, L. Gil, C. Malo, M. Olaciregui, N. González, and I. de Blas, “Sage (Salvia officinalis) and fennel (Foeniculum vulgare) improve cryopreserved boar epididymal semen quality study,” Cryoletters, vol. 36, pp. 67-70, 2013.
[26] A. Salah, H. M.M Al-Chalabi, R.M. Shukri, L. I. Mahmood, B.K. Al-Anbar, “Effect of Salvia officinalis L. (Sage) aqueous extract on liver and testicular function of diabetic albino male rats,” Journal of Babylon University/Pure and Applied Sciences, vol. 24, pp. 83-90, 2016.
[27] B. H. Ismail, and S. M. Hammed, “Effect of Salvia officinalis on the histological parameters and physiological criteria of male reproductive system in mice,” Journal of Veterinary Science, vol. 6, pp. 157-162.
[28] E. Jasem, J. Nasim, M. Gholamreza, S. Naser, M. Nader, S.L. Maryam, N. Abbas, and R. Vahid, “Evaluation of the effects of Salvia hypoleuca on the cAMP-responsive element modulator (CREM) gene expression and spermatogenesis in rat,” Middle East Fertility Society Journal, vol. 15, pp. 274-277, 2010.
[29] S. I. Moskovstev, and C. L. Librach, “Methods of sperm vitality assessment,” in: Spermatogenesis, Methods and Protocols, 1st ed. vol. 927, D. T. Carrel, and K. I. Aston, Ed. New York: Springer Science + Business Media, pp. 13-19.
[30] E. Tvrdá, N. Lukáč, J. Lukáčová, T. Jambor, and P. Massányi, “Dose- and time-dependent in vitro effects of divalent and trivalent iron on the activity of bovine spermatozoa,” Biological Trace Element Research, vol. 167, pp. 36-47, 2015.
[31] A. H. Kashou, R. Sharma, and A. Agarwal, “Assessment of oxidative stress in sperm and semen,” Methods in Molecular Biology, vol. 927, 351-361, 2013.
[32] E. Tvrdá, E. Tušimová, A. Kováčik, D. Paál, H. Greifová, A. Abdramanov, and N. Lukáč, “Curcumin has protective and antioxidant properties on bull spermatozoa subjected to induced oxidative stress,” Animal Reproduction Science, 2016, in press.
[33] B. A. Ball, “Oxidative stress, osmotic stress and apoptosis: impacts on sperm function and preservation in the horse,” Animal Reproduction Science, vol. 107, pp. 257-267, 2008.
[34] S. A. Aherne, J. P. Kerry, and N. M. O'Brien, “Effects of plant extracts on antioxidant status and oxidant-induced stress in Caco-2 cells,” British Journal of Nutrition, vol. 97, pp. 321-328, 2007.
[35] C.F. Lima, F. Carvalho, E. Fernandes, M. L. Bastos, P. C. Santos-Gomes, M. Fernandes-Ferreira, and C. Pereira-Wilson, “Evaluation of toxic/protective effects of the essential oil of Salvia officinalis on freshly isolated rat hepatocytes,” Toxicology In Vitro, vol 18, pp. 457-465, 2004.