Effect of Thymoquinone on Reproductive Parameter in Morphine-treated Male Mice

Document Type : Original Article


Department of Anatomical Sciences, Medical School, Kermanshah University of Medical Sciences, Kermanshah, Iran


Background: Thymoquinone as the main active component of Nigella sativa might have a various pharmacological effects such as antiapoptotic and antioxidant. Morphine is commonly used for the treatment of severe pain that can increase the generation of free radicals and affects the spermatogenesis. This study was designed to evaluate protective effects of thymoquinone against morphine-induced damages, sperm viability, count, motility, morphology and testis histology, and nitric oxide and testosterone hormone of the mice. Materials and Methods: In this experimental study, we divided 48 mice into eight groups (n = 6); various doses of thymoquinone (2, 10, and 20 mg/kg) and morphine (20 mg/kg) plus thymoquinone (2, 10, and 20 mg/kg) were administered intraperitoneally to 48 male mice for 30 consequent days. Male reproductive parameters including testis weight, testosterone hormone, serum nitric oxide, germinal thickness, sperm morphology, count, viability, and motility were analyzed and compared. Results: The results indicated that morphine administration significantly decreased germinal thickness, testis weight, testosterone level, viability, morphology, count, and motility of sperm and increased nitric oxide as compared to saline group (P < 0.05). However, increasing the dose of thymoquinone in the thymoquinone and thymoquinone plus morphine groups significantly decreases nitric oxide level (P < 0.05) while significantly boosted motility, morphology, count, viability of sperm cells, germinal thickness, and testosterone hormone in all groups as compared to morphine group (P < 0.05). Conclusion: It seems that thymoquinone administration could increase the quality some of spermatozoa and improves morphine-induced adverse effects on reproductive parameters in male mice


Gannon JR, Walsh TJ. The epidemiology of male infertility. In: Biennial Review of Infertility. Vol. 41. Urol Clin North Am 2015. p. 195-204.  Back to cited text no. 1
Salahshoor MR, Khashiadeh M, Roshankhah S, Kakabaraei S, Jalili C. Protective effect of crocin on liver toxicity induced by morphine. Res Pharm Sci 2016;11:120-9.  Back to cited text no. 2
Jalili C, Ahmadi S, Roshankhah S, Salahshoor M. Effect of Genistein on reproductive parameter and serum nitric oxide levels in morphine-treated mice. Int J Reprod Biomed (Yazd) 2016;14:95-102.  Back to cited text no. 3
Takzare N, Samizadeh E, Shoar S, Majidi Zolbin M, Naderan M, Lashkari A, et al. Impacts of morphine addiction on spermatogenesis in rats. Int J Reprod Biomed (Yazd) 2016;14:303-8.  Back to cited text no. 4
Vicente-Carrillo A, Álvarez-Rodríguez M, Rodríguez-Martínez H. The mu (μ) and delta (δ) opioid receptors modulate boar sperm motility. Mol Reprod Dev 2016;83:724-34.  Back to cited text no. 5
Ghowsi M, Yousofvand N. Impact of morphine dependency and detoxification by methadone on male's rat reproductive system. Iran J Reprod Med 2015;13:275-82.  Back to cited text no. 6
Jin JM, Yang WX. Molecular regulation of hypothalamus-pituitary-gonads axis in males. Gene 2014;551:15-25.  Back to cited text no. 7
Kim NN, Shin HS, Choi YJ, Choi CY. Kisspeptin regulates the hypothalamus-pituitary-gonad axis gene expression during sexual maturation in the cinnamon clownfish, Amphiprion melanopus. Comp Biochem Physiol B Biochem Mol Biol 2014;168:19-32.  Back to cited text no. 8
Ciarrocca M, Capozzella A, Tomei F, Tomei G, Caciari T. Exposure to cadmium in male urban and rural workers and effects on FSH, LH and testosterone. Chemosphere 2013;90:2077-84.  Back to cited text no. 9
Daniell HW. Hypogonadism in men consuming sustained-action oral opioids. J Pain 2002;3:377-84.  Back to cited text no. 10
Salahshoor MR, Khazaei M, Jalili C, Keivan M. Crocin improves damage induced by nicotine on a number of reproductive parameters in male mice. Int J Fertil Steril 2016;10:71-8.  Back to cited text no. 11
Niki E, Yoshida Y, Saito Y, Noguchi N. Lipid peroxidation: Mechanisms, inhibition, and biological effects. Biochem Biophys Res Commun 2005;338:668-76.  Back to cited text no. 12
Newman DJ, Cragg GM. Natural products as sources of new drugs from 1981 to 2014. J Nat Prod 2016;79:629-61.  Back to cited text no. 13
Nejatbakhsh F, Shirbeigi L, Rahimi R, Abolhassani H. Review of local herbal compounds found in the Iranian traditional medicine known to optimise male fertility. Andrologia 2016;48:850-9.  Back to cited text no. 14
Akram Khan M, Afzal M. Chemical composition of Nigella sativa Linn: Part 2 Recent advances. Inflammopharmacology 2016;24:67-79.  Back to cited text no. 15
Jalili C, Salahshoor MR, Hoseini M, Roshankhah S, Sohrabi M, Shabanizadeh A. Protective effect of thymoquinone against morphine injuries to kidneys of mice. Iran J Kidney Dis 2017;11:142-50.  Back to cited text no. 16
Paseban M. Effect of hydro-alcoholic extract of Nigella sativa on gastric acid and mucus secretion in cyclooxygenase inhibition condition in rats. Horiz Med Sci Persian 2014;20:109-13.  Back to cited text no. 17
Sheikhbahaei F, Khazaei M, Rabzia A, Mansouri K, Ghanbari A. Protective effects of thymoquinone against methotrexate-induced germ cell apoptosis in male mice. Int J Fertil Steril 2016;9:541-7.  Back to cited text no. 18
Jalili C, Salahshoor MR, Naderi T. The effect of hydroalcoholic extract of P. crispum on sperm parameters, testis tissue and serum nitric oxide levels in mice. Adv Biomed Res 2015;4:40.  Back to cited text no. 19
Slegtenhorst-Eegdeman KE, de Rooij DG, Verhoef-Post M, van de Kant HJ, Bakker CE, Oostra BA, et al. Macroorchidism in FMR1 knockout mice is caused by increased Sertoli cell proliferation during testicular development. Endocrinology 1998;139:156-62.  Back to cited text no. 20
Tawfeek FK, Kakel SJ. Effect of Nigella sativa oil treatment on the sex organs and sperm charactors in rats exposed to hydrogen peroxide. Mesopotamia J Agric 2006;34:1-7.  Back to cited text no. 21
Mabrouk A, Ben Cheikh H. Thymoquinone supplementation ameliorates lead-induced testis function impairment in adult rats. Toxicol Ind Health 2016;32:1114-21.  Back to cited text no. 22
Kaushal N, Bansal MP. Dietary selenium variation-induced oxidative stress modulates CDC2/cyclin B1 expression and apoptosis of germ cells in mice testis. J Nutr Biochem 2007;18:553-64.  Back to cited text no. 23
Heaton MB, Paiva M, Siler-Marsiglio K. Ethanol influences on bax translocation, mitochondrial membrane potential, and reactive oxygen species generation are modulated by Vitamin E and brain-derived neurotrophic factor. Alcohol Clin Exp Res 2011;35:1122-33.  Back to cited text no. 24
Rahman SA, Anwar NA, Basha SS, Kamarzaman S, Shah JS, Mahkota BI. Impact of thymoquinone supplementation on immobilisation stress-induced changes in reproductive characteristics of male mice. J App Nat Sci 2014;1:1-5.  Back to cited text no. 25
Abdallah MA, Kelany ME. Could thymoquinone protect the liver against indomethacin toxicity. Life Sci J 2014;11:26-33.  Back to cited text no. 26
Khan UA, Aslam M, Saeed SA. Effect of beta adrenergic antagonist on the production of testosterone by rat's Leydig cells. J Ayub Med Coll Abbottabad 2004;16:26-8.  Back to cited text no. 27
Kalra SP, Kalra PS. Opioid-adrenergic-steroid connection in regulation of luteinizing hormone secretion in the rat. Neuroendocrinology 1984;38:418-26.  Back to cited text no. 28
Yilmaz B, Konar V, Kutlu S, Sandal S, Canpolat S, Gezen MR, et al. Influence of chronic morphine exposure on serum LH, FSH, testosterone levels, and body and testicular weights in the developing male rat. Arch Androl 1999;43:189-96.  Back to cited text no. 29
Tawfeek FK, Ahmed SM, Kakel SJ. Effect of Nigella sativa oil treatment on the sex organs and sperm characters in rats exposed to hydrogen peroxide. Mesopotamia J Agric 2006;34:2-8.  Back to cited text no. 30
El Khasmi M, Allah AI, Farh M, Riad F, Safwate A, El Abbadi N, et al. Effect of Nigella sativa fixed oil on the hormonal profile of androgens and circulating in male rats. Phytotherapie 2011;9:338-42.  Back to cited text no. 31
Pinilla L, González LC, Tena-Sempere M, Bellido C, Aguilar E. Effects of systemic blockade of nitric oxide synthases on pulsatile LH, prolactin, and GH secretion in adult male rats. Horm Res 2001;55:229-35.  Back to cited text no. 32
Weinberg JB, Doty E, Bonaventura J, Haney AF. Nitric oxide inhibition of human sperm motility. Fertil Steril 1995;64:408-13.  Back to cited text no. 33
Stefano GB, Liu Y, Goligorsky MS. Cannabinoid receptors are coupled to nitric oxide release in invertebrate immunocytes, microglia, and human monocytes. J Biol Chem 1996;271:19238-42.  Back to cited text no. 34
Jalili C, Salahshoor MR, Naseri A. Protective effect of Urtica dioica L against nicotine-induced damage on sperm parameters, testosterone and testis tissue in mice. Iran J Reprod Med 2014;12:401-8.  Back to cited text no. 35
Guvvala PR, Sellappan S, Parameswaraiah RJ. Impact of arsenic(V) on testicular oxidative stress and sperm functional attributes in Swiss albino mice. Environ Sci Pollut Res Int 2016;23:18200-10.  Back to cited text no. 36
Cho Ping N, Hashim NH, Hasan Adli DS. Effects of Nigella sativa (Habbatussauda) oil and nicotine chronic treatments on sperm parameters and testis histological features of rats. J Evid Based Complement Altern Med 2014;22;59-65.  Back to cited text no. 37
Alhimaidi AR. Thymoqunone treatment of intracytoplasmic sperm injection (ICSI) compared to in vitro fertilization (IVF) of mice oocytes and their development in vitro. Adv Mol Med 2005;1:119-23.  Back to cited text no. 38
Gil-Guzman E, Ollero M, Lopez MC, Sharma RK, Alvarez JG, Thomas AJ Jr., et al. Differential production of reactive oxygen species by subsets of human spermatozoa at different stages of maturation. Hum Reprod 2001;16:1922-30.  Back to cited text no. 39
Bolelli G, Lafisca S, Flamigni C, Lodi S, Franceschetti F, Filicori M, et al. Heroin addiction: Relationship between the plasma levels of testosterone, dihydrotestosterone, androstenedione, LH, FSH, and the plasma concentration of heroin. Toxicology 1979;15:19-29.  Back to cited text no. 40
Kamarzaman S, Rahman SA. Effects of thymoquinone supplementation on cyclophosphamide toxicity of mouse embryo In vitro. Glob Vet 2014;12:80-90.  Back to cited text no. 41