Document Type : Original Article


1 PhD Candidate, Department of Clinical Sciences, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran

2 Department of Clinical Sciences, Faculty of Veterinary Medicine, University of Tabriz, Tabriz, Iran


Canine seminal plasma contains antioxidant enzymes to protect sperm against internally generated ROS. These enzymes are removed from seminal plasma during the process of cryopreservation. The freezing/thawing process can cause some morphological and functional changes via ice crystallization and osmolality imbalance. The present study was conducted to evaluate the effects of curcumin supplementation on sperm total count, motility, progressive motility, viability, morphology, total antioxidant capacity (TAC), DNA integrityand NOX5 gene expression of dog frozen semen. The pooled semen was allocated to fresh (Group 1) and frozen (Group 2) controls, curcumin (2.50 mM) (Group 3) and curcumin (5.00 mM), (Group 4). Sperm parameters including total sperm count, morphology, motility, progressive motility, sperm concentration and DNA integrity in addition to TAC were evaluated in fresh and frozen-thawed semen samples. Real-time PCR was used to investigate NOX5 and GADPH (reference gene) genes expressions. Curcumin at 2.50 mM provided a greater protective effect on the DNA integrity compared to 5.00 mM and control groups. TAC was significantly higher in 2.50 mM group than other groups. NOX5 gene expression in curcumin 2.50 mM was higher than 5.00 mM group. In conclusion, curcumin seems to emolliate sperm parameters and to protect sperm against sperm reactive oxygen stress and increases NOX5 gene expression.


Main Subjects


    1. Szasz F, Gabor G, Solti L. Comparative study of different methods for dog semen cryopreservation and testing under clinical conditions. Acta Vet Hung 2000; 48(3):325-333.
    2. Prinosilova P, Rybar R, Zajicova A, et al. DNA integrity in fresh, chilled and frozen-thawed canine sperm. Vet Med 2012; 57(3): 133-142.
    3. Gavazza M, Gutierrez AM, Marmunti M, et al. Lipid peroxidation assays in canine fresh semen. Rev Vet 2009; 20(2): 97-102.
    4. Strzezek R, Koziorowska-Gilun M, Kowalowka M, et al. Characteristics of antioxidant system in dog semen. Pol J Vet Sci 2009; 12(1): 55-60.
    5. Lin JK, Pan MH, Lin-Shiau SY. Recent studies on the biofunctions and biotransformations of curcumin. Biofactors 2000; 13(1-4): 153-158.
    6. Sharaf HA, Morsy FA, Shaffie NM et al. Histological and histochemical study on the protective effect of curcumin on ultraviolet irradiation induced testicular damage in albino rats. J Cytol Histol 2012; 3(6): 159-166.
    7. LoTempio MM, Veena MS, Steele HL, et al. Curcumin suppresses growth of head and neck squamous cell carcinoma. Clin Cancer Res 2005; 11(19): 6994-7002.
    8. Omur AD, Coyan K. 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. Vet Med 2016; 61(1):10-16.
    9. Sarkar FH, Li Y. Cell signaling pathways altered by natural chemopreventive agents. Mutat Res 2004; 555(1-2): 53-64.
    10. Banfi B, Molnar G, Maturana A, et al. A Ca (2+)-activated NADPH oxidase in testis, spleen, and lymph nodes. J Biol Chem 2001; 276(40): 37594-37601.
    11. Sabeur K, Ball BA. Characterization of NADPH oxidase 5 in equine testis and spermatozoa. Reproduction 2007; 134(2): 263-270.
    12. Fulton DJ. Nox5 and the regulation of cellular function. Antioxid Redox Signal 2009; 11(10): 2443-2452.
    13. Chen SJ, Allam JP, Duan YG, et al. Influence of reactive oxygen species on human sperm functions and fertilizing capacity including therapeutical approaches. Arch Gynecol Obstet 2013; 288(1): 191-199.
    14. Kutzler MA. Semen collection in the dog. Theriogenology 2005; 64(3): 747-754.
    15. Andersen K. Fertility of frozen dog semen. Acta Vet Scand 1972; 13(1): 128-130.
    16. World Health Organization. WHO laboratory manual for the examination of human semen and sperm-cervical mucus interaction. 4th ed. Cambridge, UK: Cambridge University Press, 1999; 14-22.
    17. Tejada RI, Mitchell JC, Norman A, et al. A test for the practical evaluation of male fertility by acridine orange (AO) fluorescence. Fertil Steril 1984; 42(1): 87-91.
    18. Benzie IF, Strain JJ. The ferric reducing ability of plasma (FRAP) as a measure of "antioxidant power: the FRAP assay. Anal Biochem, 1996; 239(1): 70-76.
    19. Jameel T. Sperm swim-up: A simple and effective technique of semen processing for intrauterine insemination. J Pak Med Assoc 2008; 58(2): 71-74.
    20. Almeida IL, Durfey CL, Gastal GDA, et al. 134 relaxin and its receptors in mature canine spermatozoa.Reprod Fertil Dev 2016; 29(1):175-175.
    21. Salavati M, Ghafari F, Zhang T, et al. Effects of oxygen concentration on in vitro maturation of canine oocytes in a chemically defined serum-free medium. Reproduction 2012; 144(5): 547-556.
    22. Livak JK, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta CT) method. Methods 2001; 25(4): 402-408.
    23. Izadpanah M, Alizadeh R, Minaee MB, et al. The effects of curcumin on sperm parameters and nitric oxide production in varicocelized rats.Int J Morphol 2015; 33(4): 1530-1535.
    24. Tvrda E, Lukac N, Jambor T, et al. Curcumin in male fertility: Effects on spermatozoa vitality and oxidative balance. J Mircobiol Biotech Food Sci 2015; 4(2):120-124.
    25. Soleimanzadeh A, Saberivand A. Effect of curcumin on rat sperm morphology after the freeze-thawing process. Vet Res Forum 2013; 4(3): 185-189.
    26. Cohly HH, Taylor A, Angel MF, et al. Effect of turmeric, turmerin and curcumin on H2O2-induced renal epithelial (LLC-PK1) cell injury. Free Radic Biol Med 1998; 24(1): 49-54.
    27. Salahshoor MR, Jalili C, Khazaei M, et al. Effects of curcumin on reproductive parameters in male mice. J Clin Res Paramed Sci 2012; 1(3): 1-3.
    28. Xia X, Cai H, Qin S, et al. Histone acetylase inhibitor curcumin impairs mouse spermiogenesis – an in vitro study. PLoS One 2012; 7(11): e48673.
    29. Sreejayan N, Rao MNA. Curcuminoids as potent inhibitors of lipid-peroxidation. J Pharmacol 1994; 46(12): 1013-1016.
    30. Singh S, Aggarwal BB. Activation of transcription factor NF-kB is suppressed by curcumin (Diferuloylmethane). J Biol Chem 1995; 270(42): 24995-25000
    31. Naz RK. The effect of corcumin on intracellular pH (pHi), membrane hyperpolarization and sperm motility. J Reprod Infertil 2014; 15(2): 62-70.
    32. Salles N, Szanto I, Herrmann F, et al. Expression of mRNA for ROS generating NADPH oxidases in the aging stomach. Exp Gerontol2005; 40(4): 353-357.
    33. Ghani E, Keshtgar S, Habibagahi M, et al. Expression of NOX5 in human teratozoospermia compared to
      normozoospermia. Andrologia 2013; 45(5): 351-356.
    34. Aitken RJ, Ryan AL, Baker MA, et al. Redox activity associated with the maturation and capacitation of mammalian spermatozoa. Free Radic Biol Med2004; 36(8): 994-1010.
    35. Matsuura R, Takeuchi T, Yoshida A. Preparation and incubation conditions affect the DNA integrity of ejaculated human spermatozoa. Asian J Androl 2010; 12(5): 753-759.
    36. Hong J, Behar J, Wands J, et al. Bile acid reflux contributes to development of esophageal adeno-carcinoma via activation of phosphatidylinositol-specific phospholipase Ca2+ and NADPH oxidase NOX5-S. Cancer Res2010; 70(3): 1247-1255.