Document Type : Original Article

Authors

1 Department of Theriogenology, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran

2 Department of Basic Sciences, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran

Abstract

Busulfan is known to cause several adverse effects including reproductive toxicity in humans. Garlic (Allium sativum), a widely distributed medicinal plant, is highly regarded for its medicinal activities including antioxidant property.This study was conducted to assess whether garlic extract could serve as protective agents against testicular toxicity during busulfan treatment in a mice model.Seventy-two adult male mice were randomly divided into nine groups. In groups 1,2 and 3, distilled water, busulfan, and dimethyl sulfoxide and in the treatment groups hydro-alcoholic extract of garlic was administered orally at different doses per day (groups 4, 5 and 6; 200, 400, 800 mg kg-1 respectively). Groups 7, 8 and 9 were treated with the extract (200, 400 and 800 mg kg-1, respectively) plus busulfan. Following euthanasia, blood samples and epididymal sperm were collected.The busulfan-treated group showed significant decreases in sperm qualityparameters, and serum levels of testosterone, LH and FSH was observed in the busulfan-treated mice. In addition, the TAC levels and antioxidant enzymes activities were reduced and malondialdehyde (MDA) levels were increased in the busulfan-treated mice. Notably, garlic extract co-administration caused a considerable recovery in sperm qualityparameters, TAC levels, antioxidant enzymes activities, hormonal changes and MDA level. Based on our results, garlic has antioxidant effects against busulfan-induced testicular damages in mice.

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    1. Suriapraba E, Mani PR, Kumar RA, et al. Protective effect of NAC on busulfan induced clastogenesis in human peripheral blood lymphocytes. Int J Inst Pharm Life Sci 2012; 2(1): 172-177.
    2. Sweetman SC. Martindale. London, UK: Pharmaceutical Press 2002; 33.
    3. McClive PJ, Sinclair AH. Type II and type IX collagen transcript isoforms are expressed during mouse testis development. Biol Reprod 2003; 68(5): 1742-1747.
    4. Kanatsu-Shinohara M, Toyokuni S, Morimoto T, et al. Functional assessment of self-renewal activity of male germline stem cells following cytotoxic damage and serial transplantation. Biol Reprod 2003; 68: 1801-1807.
    5. Probin V, Wang Y, Zhou D. Busulfan-induced senescence is dependent on ROS production upstream of the MAPK pathway. Free Radic Biol Med 2007; 42(12):1858-1865.
    6. Fang YZ, Yang S, Wu G. Free radicals, antioxidants, and nutrition. Nutrition 2002; 18(10): 872-879.
    7. Maymon BB, Yogev L, Marks A, et al. Sertoli cell inactivation by cytotoxic damage to the human testis after cancer chemotherapy. Fertil steril 2004; 81(5): 1391-1394.
    8. Valko M, Leibfritz D, Moncol J, et al. Free radicals and antioxidants in normal physiological functions and human disease. Int J Biochem Cell Biol 2007; 39(1): 44-84.
    9. Saxena R, Garg P, Jain DK. In vitro anti-oxidant effect of vitamin E on oxidative stress induced due to pesticides in rat erythrocytes. Toxicol Int 2011; 18(1): 73-76.
    10. Bozin B, Mimica-Dukic N, Samojlik I, et al. Phenolics as antioxidants in garlic (Allium sativum L., Alliaceae). Food Chem 2008; 111(4): 925-929.
    11. Hammami I, Nahdi A, Mauduit C, et al. The inhibitory effects on adult male reproductive functions of crude garlic (Allium sativum) feeding. Asian J Androl 2008; 10(4): 593–601.
    12. Abid-Essefi S, Zaied C, Bouaziz C, et al. Protective effect of aqueous extract of Allium sativum against zearalenone toxicity mediated by oxidative stress. Exp Toxicol Pathol 2012; 64(7): 689-695.
    13. Song K, Milner JA. The influence of heating on the anticancer properties of garlic. J Nutr 2001; 131(3s): 1054S-1057S.
    14. Santos J, Almajano MP, Carbo R. Antimicrobial, and antioxidants activity of crude onion (Allium cepa L.) extracts. Int J Food Sci Technol 2010; 45(2): 403-409.
    15. Hosseini N. Khaki A. Effect of aqueous extract of garlic (Allium sativum) on sperms morphology, motility, concentration and its antioxidant activity in rats. Afinidad 2014; 80(566): 201-204.
    16. Ghalehkandi JG. Garlic (Allium sativum) juice protects from semen oxidative stress in male rats exposed to chromium chloride. Anim Reprod 2014; 11(4): 526-532.
    17. Erdemoglu N, Kupeli E, Yesilada E. Anti-inflammatory and anti-nociceptive activity assessment of plants used as remedy in Turkish folk medicine. J Ethnopharmacol 2003; 89(1): 123-129.
    18. Homafar MA, Soleimanirad J, Ghanbari AA. A morphologic and morphometric study of adult mouse testis following different doses of busulfan administration. J Reprod Infertil 2006; 7(1):25-36.
    19. Ahmadi A, Bamohabat Chafjiri S, Sadrkhanlou R. Effect of Satureja khuzestanica essential oil against fertility disorders induced by busulfan in female mice. Vet Res Forum 2017; 8(4): 281-286.
    20. Mirfardi M, Johari H. The effect of hydro-alcoholic Allium sativum extracts on sexual hormones in mature male rats under chemotherapy with cyclo-phosphamide. Zahedan J Res Med Sci 2015; 15: 29-33.
    21. Zobeiri F, Sadrkhanlou RA, Salami S, et al. The effect of ciprofloxacin on sperm DNA damage, fertility potential and early embryonic development in NMRI mice. Vet Res Forum 2012; 3: 131-135.
    22. World Health Organization. WHO laboratory manual for the examination of human semen and sperm-cervical mucus interaction. 3rd ed. Cambridge, UK: Cambridge University Press 1992; 45-100.
    23. Wyrobek AJ, Gordon LA, Burkhart JG, et al. An evaluation of the mouse sperm morphology test and other sperm tests in nonhuman mammals. Mutat Res 1983; 115: 1-72.
    24. Narayana K, D'Souza UJ, Seetharama Rao KP. Ribvirin induced sperm shape abnormalities in Wistar rat. Mutal Res 2002; 513(1): 193-196.
    25. Bungum M, Humaidan P, Spano M, et al. Predictive value of sperm chromatin structure assay (SCSA) Para-meters for the outcome of intrauterine insemination, IVF and ICSI. Hum Reprod 2004; 19(6): 1401-1408.
    26. Hodjat M, Akhondi MA, Amirjanati N, et al. The comparison of four different sperm chromatin assays and their correlation with semen parameters. Tehran Uni Med J 2008; 65(Supl3): 33-40.
    27. Meistrich ML, Brock WA, Grimes SR, et al. Nuclear protein transitions during spermatogenesis. Fed Porc 1978; 37(11): 2522-2525.
    28. Talebi AR, Sarcheshmeh AA, Khalili MA, et al. Effect of ethanol consumption on chromatin condensation and DNA integrity of epididymal spermatozoa in rat. Alcohol 2011; 45(4): 403-409.
    29. Deiss WP. Hormone assays and their clinical application. J Am Med Assoc 1967; 200(2):184-185.
    30. Benzie IF, Strain JJ. Ferric reducing/antioxidant power assay: Direct measure of total antioxidant activity of biological fluids and modified version for simultaneous measurement of total antioxidant power and ascorbic acid concentration. Methods Enzymol 1999; 299: 15-27.
    31. Sinha AK. Colorimetric assay of catalase. Anal Biochem 1972; 47(2): 389-394.
    32. Paglia DE, Valentine WN. Studies on the quantitative and qualitative characterization of erythrocyte glutathione peroxidase. J Lab Clin Med 1967; 70(1): 158-169.
    33. Nishikimi M, Rao NA, Yogi K. Colorimetric determination of super oxide dismutase. Biochem Bioph Common 1972; 46: 849-854.
    34. Rajesh MG, Latha MS. Protective activity of Glycyrrhiza glabra Linn. on carbon tetrachloride-induced peroxidative damage. Indian J Pharmacol 2004; 36(5): 284-287.
    35. Hosseinzadeh H, Sadeghnia HR. Safranal, a constituent of Crocus sativus (saffron), attenuated cerebral ischemia induced oxidative damage in rat hippocampus. J Pharm Pharm Sci 2005; 8(3): 394-399.
    36. Mohammad-Ghasemi F, Soleimanirad J, Ghanbari AA. An ultrastructural study on the apoptotic features of spermatogenic cells following busulfan treatment in adult mice. Reprod Infertil 2008; 8(4): 319-329.
    37. Nasimi P, Tabandeh MR, Vahdati A, et al. Busulfan induces oxidative stress-and Bcl-2 family gene-related apoptosis in epididymal sperm and testis of adult male mice. Physiol Pharmacol 2015; 19(3): 208-215.
    38. Dehghani F, Hassanpour A, Poost-Pasand A, et al. Protective effects of L-carnitine and homogenized testis tissue on the testis and sperm parameters of busulfan-induced infertile male rats. Iran J Reprod Med 2013; 11(9): 693-704.
    39. Ghosh D, Das UB, Ghosh S, et al. Testicular gametogenic and steroidogenic activities in cyclophosphamide treated rat: A correlative study with testicular oxidative stress. Drug Chem Toxicol 2002; 25: 281-292.
    40. Bordbar H, Esmaeilpour T, Dehghani F, et al. Stereological study of the effect of ginger's alcoholic extract on the testis in busulfan-induced infertility in rats. Iran J Reprod Med 2013; 11(6): 467-472.
    41. Gerl A, Mühlbayer D, Hansmann G, et al. The impact of chemotherapy on Leydig cell function in long term survivors of germ cell tumors. Cancer 2001; 91(7): 1297-1303.
    42. Ilbey YO, Ozbek E, Simsek A, et al. Potential chemoprotective effect of melatonin in cyclophosphamide-and cisplatin-induced testicular damage in rats. Fertil Steril 2009; 92(3): 1124-1132.
    43. Banerjee SK, Mukherjee PK, Maulik SK. Garlic as an antioxidant: The good, the bad and the ugly. Phytother Res 2003; 17(2): 97-106.
    44. Charles DJ. Antioxidant properties of spices, herbs and other sources. New York, USA: Springer Science & Business Media 2012; 612-613.
    45. Ruwanpura SM, McLachlan RI, Meachem SJ. Hormonal regulation of male germ cell development. J Endocrinol 2010; 205(2): 117-131.
    46. Hajiuon B, Elahizadeh H. Effects of garlic (Allium sativum L.) hydro alcoholic extract on estrogen, progesterone and testosterone levels in rats exposed to cell phone radiation. Armaghane Danesh 2014; 19(5): 390-400.
    47. Zarei L, Shahrooz R, Sadrkhanlou R, et al. Protective effects of cornus mas extract on in vitro fertilization potential in methotrexate treated male mice. Vet Res Forum 2015; 6(1): 55-61.
    48. Asadpour R, Azari M, Hejazi M, et al. Protective effects of garlic aqueous extract (Allium sativum), vitamin E, and N-acetylcysteine on reproductive quality of male rats exposed to lead. Vet Res Forum 2013; 4(4): 251-257.
    49. Mirfard M, Johari H, Mokhtari M, et al. The effect of hydro-alcoholic garlic extract on testis weight and spermatogenesis in mature male rats under chemotherapy with cyclophosphamide. J Fasa Uni Med Sci 2011; 1(3):123-130.
    50. Oi Y, Imafuku M, Shishido C, et al. Garlic supplementation increases testicular testosterone and decreases plasma corticosterone in rats fed a high protein diet. J Nutr 2001; 131(8): 2150-2156.