Effect of silymarin antioxidant on the membrane integrity, viability and motility of ram frozen sperm

Document Type : Research Paper

Authors

1 MSc student of Animal Science, Faculty of Agricultre, University of Tabriz

2 Department of Animal Science, Faculty of Agricultre, University of Tabriz

Abstract

Effect of silymarin antioxidant on the membrane integrity, viability and motility of ram frozen sperm

Abstract:
Introduction: The need for artificial insemination in the ewe is to access fresh and frozen sperm. Cryopreservation is a suitable method for long-term storage of semen and sperm, but may cause a minor irreversible damage to the sperm cell, especially its membrane region (Khorramabadi et al., 2017). The oxidation of fatty acids leads to the production of free oxygen radicals (ROS). These radicals are necessary in normal conditions for certain physiological activities and sperm processes, but excessive production of ROS in the sperm can reduce membrane fluidity, DNA fracture, damage to proteins, and ultimately reduced sperm motility and fertility. (Bakhshayesh et al., 2017). Silymarin, the scientific name of Silbum marianum, is the English name Milk Thistle and is a potent inhibitor of oxidative stress as a potent antioxidant. Due to the nature of its friend fat, silymarin is firmly attached to the plasmid membrane compounds, thus preventing fracture and its collapse by increasing membrane strength. Increasing the amount of active oxygen species and lipid peroxidation disrupts mitochondrial membrane, decreases ATP and damage to sperm acesomes, which ultimately reduces the progression of sperm motility. (Kvasnikova et al., 2003). The mechanism of action of salimarin is through stimulation of the ribosomal RNA and protects the membrane from oxidative damage. They also stated that silymarin stimulates the activity of the antioxidant enzymes of superoxide dismutase (SOD) and glutathione peroxidase (Wellington and Jarvis, 2001) . Silymarin improves the sperm motility and survival, sperm abnormalities, and preservation of the sperm membrane after freezing-thawing. Improvement of semen quality in both freezing and cooling is due to the strong silicomain antioxidant capacity (Fakorzai et al., 2008; Longpyram et al., 2013). . Objective: Regarding the anti-oxidant effects of silymarin, this study aimed to investigate the effect of silymarin on storage of ram sperm.
Materials and Methods: In this research, four rams of pure ghee were used 2-3 years old and each ram was 6 times sperm. Sperm was performed twice a week by artificial vagina. After the sample was transferred to the laboratory, a series of preliminary evaluations including sample size, sample color, wave motion, total mobility, in-situ motion, progressive motion and live weight, and abnormal sperm and density were performed. If standards were met, Necessary (scaling over 2.5 billion sperm and progressive movement above 70%) dilution was performed with treatments. The diluents were prepared prior to sampling and placed in a bin Marie temperature of 37 ° C. Tris-based diluent was used to contain 2.73 g of tries, 1.4 g of fructose, 1 g of citric acid and 100 mg streptomycin in 100 ml sterile distilled water. To prepare the diluent, 73% of the prepared solution will be mixed with 20% egg yolk and 7% glycerol. In order to dilute the sprays from trace based diluent, egg yolk, semen collected with diluent without antioxidants (control) and silymarin 5 and 10 μg / ml mix After cooling for 90 minutes in the refrigerator and reaching 5 ° C, it was placed in 4 cm above the liquid nitrogen for 8-10 minutes and then immersed in liquid nitrogen. The post-mortem examinations of sperm quality properties included liveliness, total motion, progressive motion, integrity of the plasma membrane (tests) on days 0, 15, 30 of the experiment.
Results: According to the results of the experiment, the diluents containing the levels of silymarin significantly increased the total motion, progressive, live survival of ram sperm after the freeze-thawing process compared to the control group Were opened. Addition of different concentrations of silymarin was 5 and 10 μg with mean of 74.45% and 67.33%, respectively, compared to the control group, significantly increased the total sperm motility with mean of 68.07 and 60.95%, significantly Progressive motility of frozen ram sperm were frozen (p <0.01). According to the results, the in situ movement in the control group (5.92 ± 0.92) was lower than that of the 5 and 10 μg silymarin (6.38 ± 0.92) and (8.9 ± 0.92) respectively In the least amount.
Conclusion: Application of silymarin as an antioxidant in the ram sperm diluent system reduces oxidative damage and improves sperm quality properties such as mobility, viability and the health of acrosome and plasma membrane during freezing process. Was opened. Adding 5 μg of silymarin to diluted ram minus 10 μg and control group reduced fat peroxidation and also improved the structure and performance of sperm during storage.

Keywords


Ahmadi Hamedani M, Jafari Ahangari Y and Zerehdaran S, 2014. Effect of Different Levels of Egg Yolk on Tris Dilution in Sperm Cow's Rat Quality in Cooling and Freezing Conditions. Journal of Animal Production Research 5(10)122-134.
Ahmadi Hamedani M, Tahmasebi A, Nasserian A and Jafari Bohrani, 2016. Effect of vitamin B12 in Tris based diluent on the protection of sperm rams. Journal of Research in Ruminants 4(3)73-92.
Aitken J and Fisher H, 1994. Reactive oxygen species generation and human ‎spermatozoa: the balance ‎of benefit and risk. Bioassays16: 259-267. ‎
Alfati Karjei R, Daghigh kia H, Moghadam Gh, Hossein Khani A and Alijani S, 2013. Effect of Different Levels of Glutathione and Superoxide Dismutase on Some Characteristics of Cow's Sperm after Freezing. Journal of Animal and Poultry Research 1(4) 1-7.
Bailey JL, Bilodeau JF, and Cormier N, 2000. Semen cryopreservation in domestic animals: A damaging and capacitating phenomenon. Journal of Andrology 21: 1-7.
Bakhshayesh Khiabani Moghaddam G and Daghigh Kia H, 2017.  Effects of adding different levels of Glutamine to modified Beltsville extender on the survival of frozen rooster semen. Animal Reproduction Science 184: 172–177.
Baumber J, Ball BA, Gravance CG, Medina V and Morel MCG, 2000.The effect of reactive oxygen species on equine sperm motility, viability, acrosomal integrity, mitochondrial membrane potential and membrane lipid–peroxidation. Journal of Andrology 21: 895-902.
Bilodeau JF, Chatterjee S, Sirard MA and Gagnon C. 2000. Levels of antioxidant ‎defenses are decreased in bovine spermatozoa after a cycle of freezing and thawing. Molecular Reproduction and Development 55:282–288.
Blocki Z and Daghigh kia H, 2017.Improvement of ram semen quality after the freezing- thawing process using various levels of coenzyme Q10. Journal of Animal Production 19(3) 727-738.
Chatterjee S, Lamirande E and Gagnon C, 2001. Cryopreservation alters membrane ‎sulfhydryl ‎status of bull spermatozoa: protection by oxidized glutathione. Molecular Reproduction and Development 60: 498–506.
Dolati Durbash P, Moghaddam G, Daghigh kia H, Taghizadeh A and Rafat A, 2015. The Effect of Different Use of Rafinose Sugar in the Maintenance of Frozen Sperm of Different Breeds of Rams in the Reproductive Period, Journal of Animal Science Research 25(2)121-132.
El Sheshtawy R, and El-Natta W, 2017. Impact of silymarin enriched semen extender on bull sperm preservability. Asian Pacific Journal of Reproduction 6(2) 81-84.‏
Evans G and Maxwell WMC, 1987. Handling and examination semen. In: Maxwell WMC, editor. Salamons artificial insemination of sheep and goat. Sydney: Butterworths 93-106.
Fakurazi S, Nanthini   U and Hairusah I, 2008.Hepatoprotective and antioxidant action of Moringa oleifera Lam.against acetaminophen induced hepatotoxicity in rats. International Journal of Pharmacology 4(4) 270-275.
Khorram Abadi F, Khodaee Motlagh M and Moradi M, 2017. Investigating the addition of selenium nanoparticles in semen diluent in the laboratory on sperm parameters after freezing ram Farahani. Journal of Zoology Research 30(3)301-307
Kvasnicka f, Biba B, sevcik R, Voldrich M and Karta J, 2003. Analysis of the active components of silymarin. Journal of Chromatography A, 990: 239–245
Luangpiromn   A,   Junaimuang   T, Kourchampa W, Somsapt   P and   Sritragoo O, 2013. Protective effect of pomegranate (Punica granatum Linn.) ice against hepatotoxicity and testicular toxicity induced by ethanol in mice. Animal Biology and Animal Husbandry 5(1)15-19
Medeiros CMO, Forell F, Oliveira ATD and Rodrigues JL, 2002. Current status of sperm cryopreservation: why isn't it better?.  Theriogenology 57(1) 327-344.
Mohammadi N, Moghadam Gh, Daghigh kia H, Javanmard A, 2017. Protecting mitochondrial function during the process of freezing of sperm. Age of Life Publishing.
 
Momeni H  , Sepehri H and Yousefi M, 2014. The effect of silymarin on plasma membranes and acrosome of treated sperm with aluminum chloride. Journal of Arak University of Medical Sciences 18(4) 80-71.
Momeni H., Sepehri H andYousefi M. 2015. The effect of silymarin on plasma membrane and acrosome of aluminum chloride treated sperm, Journal of Arak University of Medical Sciences 4:71-80.
Momeni H., Khavari A, Khodaei motlaq M, Chobinh T and  Eskandari N,2014. The role of Silymarin on survival, mobility and integrity of cadmium chlorid treated sperm acrosomes, Iranian Journal of Animal Science Researches 9(4)498-506.
Parisuj P, 2013. Effect of different amounts of silymarin and caproic acid on storage of ram sperm at 5 ° C. End of letter. Gilan University.
Salamon S and Maxwell WMC, 2000. Storage of ram semen. Animal Reproduction Science 62(1) 77-111.
Sariozhan S, Bucack MN, Purhan BT, Pinnar AU and Ali B, 2009. The influence of cysteine and taurine on microscopic oxidative stress paramerters and fertilizing ability of bull semen following cryopreservation. Cryobiology, 58(2) 134-138.
Tabatabaei Vakili S, Zaidi Z, Mamoli M and Mirzadeh Kh, 2017. Investigating of different levels of glutamine aminoasid on the quality of Arabic ram semen in different storage times to 5° C. Journal of Animal Sciences 115: 233-242.
Wellington K and Jarvis B, 2001. Silymarin: A review of its clinical properties in the management of hepatic disorders. BioDrugs Journal, 15(7) 465-489.
Zamiri m, 2012. Reproductive physiology.Third edition, Haghshenas Publishing.
Ziaei Rad H, Roostaee Ali Mehr M and Mohammadi M, 2016. The effect of silymarin on cervical semen storage at 4 ° C. Journal of Animal Science Research 26(3)1-13.