Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 30132
In vitro Effects of Amygdalin on the Functional Competence of Rabbit Spermatozoa

Authors: Marek Halenár, Eva Tvrdá, Tomáš Slanina, Ľubomír Ondruška, Eduard Kolesár, Peter Massányi, Adriana Kolesárová

Abstract:

The present in vitro study was designed to reveal whether amygdalin (AMG) is able to cause changes to the motility, viability and mitochondrial activity of rabbit spermatozoa. New Zealand White rabbits (n = 10) aged four months were used in the study. Semen samples were collected from each animal and used for the in vitro incubation. The samples were divided into five equal parts and diluted with saline supplemented with 0, 0.5, 1, 2.5 and 5 mg/mL AMG. At times 0h, 3h and 5h spermatozoa motion parameters were assessed using the SpermVision™ computer-aided sperm analysis (CASA) system, cell viability was examined with the metabolic activity (MTT) assay, and the eosin-nigrosin staining technique was used to evaluate the viability of rabbit spermatozoa. All AMG concentrations exhibited stimulating effects on the spermatozoa activity, as shown by a significant preservation of the motility (P<0.05 with respect to 0.5 mg/mL and 1 mg/mL AMG; Time 5 h) and mitochondrial activity (P< 0.05 in case of 0.5 mg/mL AMG; P< 0.01 in case of 1 mg/mL AMG; P < 0.001 with respect to 2.5 mg/mL and 5 mg/mL AMG; Time 5 h). None of the AMG doses supplemented had any significant impact of the spermatozoa viability. In conclusion, the data revealed that short-term co-incubation of spermatozoa with AMG may result in a higher preservation of the sperm structural integrity and functional activity.

Keywords: Amygdalin, CASA, mitochondrial activity, motility, rabbits, spermatozoa, viability.

Digital Object Identifier (DOI): doi.org/10.5281/zenodo.1127330

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 761

References:


[1] D. Dushenkov, and I. Raskin, “New strategy for the search of natural biologically active substances,” Russian Journal of Plant Physiology, vol. 55, pp. 564-567, 2008.
[2] S. Sasidharan, Y. Chen, D. Saravanan, K. M. Sundram, and L. Yoga Latha, “Extraction, isolation and characterization of bioactive compounds from plants' extracts,” African Journal of Traditional, Complementary and Alternative Medicines, vol. 8, pp. 1-10, 2011.
[3] M. Halenar, M. Medvedova, N. Maruniakova, and A. Kolesarova, “Assessment of a potential preventive ability of amygdalin in mycotoxin-induced ovarian toxicity,” Journal of Environmental Science and Health, Part B, vol. 50, pp. 411-416, 2015.
[4] M. M. Ames, T. P. Moyer, J. S. Kovach, C. G. Moertel, and J. Rubin, “Pharmacology of amygdalin (Laetrile) in cancer patients,” Cancer Chemotherapy and Pharmacology, vol. 6, pp. 51-57, 1981.
[5] H. K. Chang, M. S. Shin, H. Y. Yang, J. W. Lee, Y. S. Kim, M. H. Lee, J. Kim, K. H. Kim, and C. J. Kim, “Amygdalin induces apoptosis through regulation of Bax and Bcl-2 expressions in human DU145 and LNCaP prostate cancer cells,” Biological and Pharmaceutical Bulletin, vol. 29, pp. 1597-1602, 2006.
[6] J. Yan, S. Tong, J. Li, and J. Lou, J., “Preparative isolation and purification of amygdalin from Prunus armeniaca L. with high recovery by high-speed countercurrent chromatography,” Journal of Liquid Chromatography & Related Technologies, vol. 29, pp. 1271-1279, 2006.
[7] T. Fukuda, H. Ito, T. Mukainaka, H. Tokuda, H. Nishino, and T. Yoshida, “Anti-tumor promoting effect of glycosides from Prunus persica seeds, Biological and Pharmaceutical Bulletin, vol. 26, pp. 271-273, 2003.
[8] H. J. Hwang, H. J. Lee, Ch, J, Kim, I. Shim, and D. H. Hahm, “Inhibitory effect of amygdalin on lipopolyccharide-inducible TNF-α and IL-1β mRNA expression and carrageenan-induced rats arthritis,” Journal of Microbiology and Biotechnology, vol. 18, pp. 1641-1647, 2008.
[9] J. Makarević, J. Rutz, E. Juengel, S. Kaulfuss, M. Reiter, I. Tsaur, G. Bartsch, A. Haferkamp, and R. A. Blaheta, “Amygdalin blocks bladder cancer cell growth in vitro by diminishing cyklin A and cdk2,” PLoS ONE, vol. 9, pP. 1-9, 2014.
[10] C. G. Moertel, C.G., Ames, M.M., Kovach, J.S., Moyer, T.P., Rubin, J.R., and J. H. Tinker, “A pharmacologic and toxicological study of amygdalin,” JAMA, vol. 245, pp. 591–594, 1981.
[11] F. A. Yildirim, and M. A. Askin, “Variability of amygdalin content in seeds of sweet or bitter apricot cultivars in Turkey,” African Journal of Biotechnology, vol. 9, pp. 6522–6524, 2010.
[12] 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.
[13] 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.
[14] M. Halenár, M. Medveďová, N. Maruniaková, and A. Kolesárová, “Amygdalin and its effects on animal cells,” Journal of Microbiology, Biotechnology and Food Sciences, vol. 2, pp. 1414-1423, 2013.
[15] A. Kolesárová, M. Capcarová, Z. Baková, B. Gálik, M. Juráček, M. Šimko, and A. V. Sirotkin, “The effect of bee pollen on secretion activity, markers of proliferation and apoptosis of porcine ovarian granulosa cells in vitro,” Journal of Environmental Science and Health, Part B, vol. 46, pp. 207-212, 2011.
[16] S. Tanyildizi, and T. Bozkurt, 2004. “In vitro effects of linamarin, amygdalin and gossypol acetic acid on hyaluronidase activity, sperm motility and morphological abnormality in bull sperm,” Turkish Journal of Veterinary and Animal Sciences, vol. 28, pp. 819-824, 2004.
[17] T. Yasui, T. Matsuzaki, K. Ushigoe, A. Kuwahara, M. Maegawa, H. Furumoto, T. Aono, and M. Irahara, “Stimulatory effect of the herbal medicine Keishi-bukuryo-ganon a cytokine-induced neutrophil chemoattractant, in rat ovarian cell culture,” American Journal of Reproductive Immunology, vol. 50, pp. 90-97, 2003.
[18] E. Tvrdá, E. Tušimová, A. Kováčik, D. Paál, Ľ. Libová, and N. Lukáč, “Protective effects of quercetin on selected oxidative biomarkers in bovine spermatozoa subjected to ferrous ascorbate,” Reproduction in Domestic Animals, to be published, 2016.
[19] Y. Chen, J. Ma, and F. Wang, “Amygdalin induces apoptosis in human cervical cancer cell line HeLa cells,” Immunopharmacology and Immunotoxicology, vol. 35, pp. 43-51, 2013.
[20] H. J. Park, S. H. Yoon, and L. Han, “Amygdalin inhibits genes related to cell cycle in SNU-C4 human colon cancer cells,” World Journal of Gastroenterology, vol. 11, pp. 5156-5161, 2005.
[21] J. Elia, N. Imbrogno, M. Delfino, R. Mazzilli, T. Rossi, and F. Mazzilli, “The importance of the sperm motility classes—future directions,” Open Andrology Journal, vol. 2, p. 42-43, 2010.
[22] P. Massanyi, P. Chrenek, N. Lukáč, A. V. Makarevich, A. Ostro, J. Živčák, and J. Bulla, “Comparison of different evaluation chambers for analysis of rabbit spermatozoa motility using CASA system,” Slovak Journal of Animal Science, vol. 41, pp. 60-66, 2008.
[23] N. Lukac, L. Bardos, R. Stawarz, S. Roychoudhury, A. V. Makarevich, P. Chrenek, J. Danko, and P. Massanyi, “In vitro effect of nickel on bovine spermatozoa motility and annexin V-labeled membrane changes,” Journal of Applied Toxicology, vol. 31, pp. 144-149, 2011.
[24] R. Eliasson, “Semen analysis with regard to sperm number, sperm morphology and functional aspects,” Asian Journal of Andrology, vol. 12, pp. 26-32, 2010.
[25] M. Philippe, and P. Chevaillier, “Extraction and biochemical characterization of a nuclear deoxyribonucleic acid polymerase activity in bull spermatozoa,” Biochemical Journal, vol. 175, p. 585-594, 1978.
[26] S. Waga, T. Masuda, and H. Takisawa, “DNA polymerase varepsilon is required for coordinated and efficient chromosomal DNA replication in Xenopus egg extracts,” PNAS, vol. 98, p. 4978-4983, 2001.
[27] Y. Mizushina, N. Takahashi, A. Ogewa, K. Tsurugaya, H. Koshino, M. Takemura, S. Yoshida, A. Matsukage, F. Sugawara, and K. Sakaguchi, “The cyanogenic glucoside, prunasin (Dmandelonitrile-beta-D-glucoside), is a novel inhibitor of DNA polymerase beta,” Journal of Biochemistry (Tokio), vol. 126, pp. 430-436, 1999.
[28] A. Amaral, J. Ramalho-Santos, and J. C. St John, “The expression of polymerase gamma and mitochondrial transcription factor A and the regulation of mitochondrial DNA content in mature human sperm,” Human Reproduction, vol. 22, pp. 1585-1596, 2007.
[29] A. P. Sousa, A. Amaral, M. Baptista, R. Tavares, P. Caballero Campo, P. Caballero Peregrin, A. Freitas, A. Pavia, T. Almeida-Santos, and J. Ramalho-Santos, “Not all sperm are equal: functional mitochondria characterize a subpopulation of human sperm with better fertilization potential,” PLoS ONE, vol. 6, e18112, 2011.
[30] F. G. Mann, and B. Ch. Saunders, Practical Organic Chemistry (4th ed.). London: Longman, 1975, pp. 509-517.
[31] Conchie and T. Mann, “Glycosidases in mammalian sperm and seminal plasma,” Nature, vol. 179, pp. 1190-1191, 1957.
[32] A. Jauhiainen, and T. Vanha-Perttula, “Characterization of acid and neutral alpha-mannosidases in bull semen and reproductive organs,” International Journal of Biochemistry, vol. 19, pp. 267-274, 1987.
[33] P. Piomboni, R. Focarelli, A. Stendardi, A. Ferramosca, and V. Zara, “The role of mitochondria in energy production for human sperm motility,” International Journal of Andrology, vol. 35, vol. 109-124, 2012.
[34] T. Mosmann, “Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays,” Journal of Immunological Methods, vol. 65, pp. 55-63, 1983.