Protective effects of the antioxidants curcumin, ellagic acid and methionine on motility, mitochondrial transmembrane potential, plasma membrane and acrosome integrity in freeze-thawed Merino ram sperm

https://doi.org/10.17221/8677-VETMEDCitation:Omur A.D., Coyan K. (2016): Protective effects of the antioxidants curcumin, ellagic acid and methionine on motility, mitochondrial transmembrane potential, plasma membrane and acrosome integrity in freeze-thawed Merino ram sperm. Veterinarni Medicina, 61: 10-16.
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The aim of this study was to determine the effects of curcumin, ellagic acid and methionine on sperm parameters following the freeze-thawing of Merino ram semen. Ejaculates were collected via an artificial vagina from four Merino rams, evaluated microscopically and pooled at 37 °C. The pooled semen samples were diluted in a Tris-based extender and separated into groups containing curcumin (1, 2, 4mM), ellagic acid (1, 2, 4mM), methionine (1, 2, 4mM) and no antioxidant (control). The diluted semen was cooled to 5 °C slowly and equilibrated for 3 h. After the equilibration, the samples were frozen in liquid nitrogen vapour, and plunged into liquid nitrogen (–196 °C) for storage. Frozen straws were thawed at 37 °C for 30 s in a water bath for microscopic sperm evaluation, individually. All antioxidants led to a higher percentage of sperm motility in comparison to the control group. The freezing extender supplemented with methionine (1mM), curcumin (1 and 2mM) and ellagic acid (1 and 2mM) led to higher percentage of sperm plasma membrane integrity when compared to other groups (P < 0.05). Antioxidant supplementation also resulted in a higher percentage of sperm acrosome integrity in comparison to the control. Methionine, curcumin and ellagic acid (1mM: 27.7 ± 2.4, 28.0 ± 2.1 and 26.8 ± 2.0) groups provided higher protection in terms of sperm mitochondrial activity when compared to other groups (P < 0.05). The findings of this study show that varying concentrations of curcumin, methionine and ellagic acid have markedly different effects on the spermatological variables under study.
References:
Aitken R. John (1994): Pathophysiology of human spermatozoa. Current Opinion in Obstetrics and Gynecology, 6, 128???135-  https://doi.org/10.1097/00001703-199404000-00004
 
Aitken RJ, Sawyer D (2003): The human spermatozoon – not waving but drowning. Advances in Experimental Medicine and Biology 518, 85–98.
 
AITKEN R. J., WEST K. M. (1990): Analysis of the relationship between reactive oxygen species production and leucocyte infiltration in fractions of human semen separated on Percoll gradients. International Journal of Andrology, 13, 433-451  https://doi.org/10.1111/j.1365-2605.1990.tb01051.x
 
ALVAREZ JUAN G., TOUCHSTONE JOSEPH C., BLASCO LUIS, STOREY BAYARD T. (1987): Spontaneous Lipid Peroxidation and Production of Hydrogen Peroxide and Superoxide in Human Spermatozoa Superoxide Dismutase as Major Enzyme Protectant Against Oxygen Toxicity. Journal of Andrology, 8, 338-348  https://doi.org/10.1002/j.1939-4640.1987.tb00973.x
 
Bucak Mustafa Numan, Sarıözkan Serpil, Tuncer Pürhan Barbaros, Sakin Fatih, Ateşşahin Ahmet, Kulaksız Recai, Çevik Mesut (2010): The effect of antioxidants on post-thawed Angora goat (Capra hircus ancryrensis) sperm parameters, lipid peroxidation and antioxidant activities. Small Ruminant Research, 89, 24-30  https://doi.org/10.1016/j.smallrumres.2009.11.015
 
Bucak M. N., Başpınar N., Tuncer P. B., Çoyan K., Sarıözkan S., Akalın P. P., Büyükleblebici S., Küçükgünay S. (2012): Effects of curcumin and dithioerythritol on frozen-thawed bovine semen. Andrologia, 44, 102-109  https://doi.org/10.1111/j.1439-0272.2010.01146.x
 
Çeribaşi Ali Osman, Türk Gaffari, Sönmez Mustafa, Sakin Fatih, Ateşşahin Ahmet (2010): Toxic Effect of Cyclophosphamide on Sperm Morphology, Testicular Histology and Blood Oxidant-Antioxidant Balance, and Protective Roles of Lycopene and Ellagic Acid. Basic & Clinical Pharmacology & Toxicology, 107, 730-736  https://doi.org/10.1111/j.1742-7843.2010.00571.x
 
Çoyan Kenan, Başpınar Nuri, Bucak Mustafa Numan, Akalın Pınar Peker, Ataman Mehmet Bozkurt, Ömür Ali Doğan, Güngör Şükrü, Küçükgünay Sadık, Özkalp Birol, Sarıözkan Serpil (2010): Influence of methionine and dithioerythritol on sperm motility, lipid peroxidation and antioxidant capacities during liquid storage of ram semen. Research in Veterinary Science, 89, 426-431  https://doi.org/10.1016/j.rvsc.2010.03.025
 
Esterbauer Hermann, Dieber-Rotheneder Martina, Waeg Georg, Striegl Georg, Juergens Guenther (1990): Biochemical structural and functional properties of oxidized low-density lipoprotein. Chemical Research in Toxicology, 3, 77-92  https://doi.org/10.1021/tx00014a001
 
Gadella B.M., Miller N.G.A., Colenbrander B., Van Golde L.M.G., Harrison R.A.P. (1999): Flow cytometric detection of transbilayer movement of fluorescent phospholipid analogues across the boar sperm plasma membrane: Elimination of labeling artifacts. Molecular Reproduction and Development, 53, 108-125  https://doi.org/10.1002/(SICI)1098-2795(199905)53:1<108::AID-MRD13>3.0.CO;2-K
 
Garner D. L. (1995): Viability assessment of mammalian sperm using SYBR-14 and propidium iodide. Biology of Reproduction, 53, 276-284  https://doi.org/10.1095/biolreprod53.2.276
 
Garner D. L. (1997): Fluorometric assessments of mitochondrial function and viability in cryopreserved bovine spermatozoa. Biology of Reproduction, 57, 1401-1406  https://doi.org/10.1095/biolreprod57.6.1401
 
Głombik Katarzyna, Basta-Kaim Agnieszka, Sikora-Polaczek Marta, Kubera Marta, Starowicz Gabriela, Styrna Józefa (2014): Curcumin influences semen quality parameters and reverses the di(2-ethylhexyl)phthalate (DEHP)-induced testicular damage in mice. Pharmacological Reports, 66, 782-787  https://doi.org/10.1016/j.pharep.2014.04.010
 
GOMEZ , IRVINE , AITKEN (1998): Evaluation of a spectrophotometric assay for the measurement of malondialdehyde and 4-hydroxyalkenals in human spermatozoa: relationships with semen quality and sperm function. International Journal of Andrology, 21, 81-94  https://doi.org/10.1046/j.1365-2605.1998.00106.x
 
Jaruga Ewa, Salvioli Stefano, Dobrucki Jurek, Chrul Sławomir, Bandorowicz-Pikuła Joanna, Sikora Ewa, Franceschi Claudio, Cossarizza Andrea, Bartosz Grzegorz (1998): Apoptosis-like, reversible changes in plasma membrane asymmetry and permeability, and transient modifications in mitochondrial membrane potential induced by curcumin in rat thymocytes. FEBS Letters, 433, 287-293  https://doi.org/10.1016/S0014-5793(98)00919-3
 
Levine Rodney L., Berlett Barbara S., Moskovitz Jackob, Mosoni Laurent, Stadtman Earl R. (1999): Methionine residues may protect proteins from critical oxidative damage. Mechanisms of Ageing and Development, 107, 323-332  https://doi.org/10.1016/S0047-6374(98)00152-3
 
Maia Marciane da Silva, Bicudo Sony Dimas, Sicherle Carmen Cecilia, Rodello Leandro, Gallego Isabel Cristina Saltaren (2010): Lipid peroxidation and generation of hydrogen peroxide in frozen-thawed ram semen cryopreserved in extenders with antioxidants. Animal Reproduction Science, 122, 118-123  https://doi.org/10.1016/j.anireprosci.2010.08.004
 
MATHURIA N, VERMA R (2008): Curcumin ameliorates aflatoxin-induced toxicity in mice spermatozoa. Fertility and Sterility, 90, 775-780  https://doi.org/10.1016/j.fertnstert.2007.07.1300
 
Mieusset Roger, Bujan Louis, Mondinat Charlotte, Mansat Arlette, Pontonnier Francis, Grandjean Helene (1987): Association of scrotal hyperthermia with impaired spermatogenesis in infertile men. Fertility and Sterility, 48, 1006-1011  https://doi.org/10.1016/S0015-0282(16)59600-9
 
Moein MR, Dehghani VO, Tabibnejad N, Vahidi S (2007): Reactive Oxygen Species (ROS) level in seminal plasma of infertile men and healthy donors. Iranian Journal of Reproductive Medicine 5, 51–55.
 
Nagy S. (2002): A Triple-Stain Flow Cytometric Method to Assess Plasma- and Acrosome-Membrane Integrity of Cryopreserved Bovine Sperm Immediately after Thawing in Presence of Egg-Yolk Particles. Biology of Reproduction, 68, 1828-1835  https://doi.org/10.1095/biolreprod.102.011445
 
Naz RK (2011): Can curcumin provide an ideal contraceptive? Molecular Reproduction and Development 78, 116–123.
 
Ollero M., Muiño Blanco T., López-Pérez M.J., Cebria Pérez J.A. (1996): Surface changes associated with ram sperm cryopreservation revealed by counter-current distribution in an aqueous two-phase system effect of different cryoprotectants. Journal of Chromatography B: Biomedical Sciences and Applications, 680, 157-164  https://doi.org/10.1016/0378-4347(95)00461-0
 
Pari Leelavinothan, Sivasankari Ramasamy (2008): Effect of ellagic acid on cyclosporine A-induced oxidative damage in the liver of rats. Fundamental & Clinical Pharmacology, 22, 395-401  https://doi.org/10.1111/j.1472-8206.2008.00609.x
 
Peris Soliman I., Bilodeau Jean-François, Dufour Maurice, Bailey Janice L. (2007): Impact of cryopreservation and reactive oxygen species on DNA integrity, lipid peroxidation, and functional parameters in ram sperm. Molecular Reproduction and Development, 74, 878-892  https://doi.org/10.1002/mrd.20686
 
Practice Committee of the American Society for Reproductive Medicine (2004): Report on varicocele and infertility. Fertility and Sterility 82, 142–145.
 
Rashid Kahkashan, Sil Parames C. (2015): Curcumin ameliorates testicular damage in diabetic rats by suppressing cellular stress-mediated mitochondria and endoplasmic reticulum-dependent apoptotic death. Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease, 1852, 70-82  https://doi.org/10.1016/j.bbadis.2014.11.007
 
Reed D J (1990): Glutathione: Toxicological Implications. Annual Review of Pharmacology and Toxicology, 30, 603-631  https://doi.org/10.1146/annurev.pa.30.040190.003131
 
Salamon S, Maxwell W.M.C (2000): Storage of ram semen. Animal Reproduction Science, 62, 77-111  https://doi.org/10.1016/S0378-4320(00)00155-X
 
Sanocka D, Kurpisz M (2004): Reactive oxygen species and sperm cells. Reproductive Biology and Endocrinology 2, 12. https://doi.org/10.1186/1477-7827-2-12
 
Smith John T., Mayer Dennis T. (1955): Evaluation of Sperm Concentration by the Hemacytometer Method. Fertility and Sterility, 6, 271-275  https://doi.org/10.1016/S0015-0282(16)31987-2
 
Starkov Anatoly A. (2008): The Role of Mitochondria in Reactive Oxygen Species Metabolism and Signaling. Annals of the New York Academy of Sciences, 1147, 37-52  https://doi.org/10.1196/annals.1427.015
 
Surh Young-Joon, Chun Kyung-Soo, Cha Hyun-Ho, Han Seong Su, Keum Young-Sam, Park Kwang-Kyun, Lee Sang Sup (2001): Molecular mechanisms underlying chemopreventive activities of anti-inflammatory phytochemicals: down-regulation of COX-2 and iNOS through suppression of NF-κB activation. Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis, 480-481, 243-268  https://doi.org/10.1016/S0027-5107(01)00183-X
 
Tuncer Pürhan Barbaros, Bucak Mustafa Numan, Sarıözkan Serpil, Sakin Fatih, Yeni Deniz, Çiğerci İbrahim Hakkı, Ateşşahin Ahmet, Avdatek Fatih, Gündoğan Mustafa, Büyükleblebici Olga (2010): The effect of raffinose and methionine on frozen/thawed Angora buck (Capra hircus ancryrensis) semen quality, lipid peroxidation and antioxidant enzyme activities. Cryobiology, 61, 89-93  https://doi.org/10.1016/j.cryobiol.2010.05.005
 
Türk Gaffari, Ateşşahin Ahmet, Sönmez Mustafa, Çeribaşi Ali Osman, Yüce Abdurrauf (2008): Improvement of cisplatin-induced injuries to sperm quality, the oxidant-antioxidant system, and the histologic structure of the rat testis by ellagic acid. Fertility and Sterility, 89, 1474-1481  https://doi.org/10.1016/j.fertnstert.2007.04.059
 
Vernet P, Aitken R.J, Drevet J.R (2004): Antioxidant strategies in the epididymis. Molecular and Cellular Endocrinology, 216, 31-39  https://doi.org/10.1016/j.mce.2003.10.069
 
Watanabe Hiroyuki, Fukui Yutaka (2006): Effects of dithiothreitol and boar on pronuclear formation and embryonic development following intracytoplasmic sperm injection in pigs. Theriogenology, 65, 528-539  https://doi.org/10.1016/j.theriogenology.2005.04.031
 
Whittington Kate, Ford (1999): Relative contribution of leukocytes and of spermatozoa to reactive oxygen species production in human sperm suspensions. International Journal of Andrology, 22, 229-235  https://doi.org/10.1046/j.1365-2605.1999.00173.x
 
Wright C., Milne S., Leeson H. (2014): Sperm DNA damage caused by oxidative stress: modifiable clinical, lifestyle and nutritional factors in male infertility. Reproductive BioMedicine Online, 28, 684-703  https://doi.org/10.1016/j.rbmo.2014.02.004
 
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