International Journal of Reproductive BioMedicine
ISSN: 2476-3772
The latest discoveries in all areas of reproduction and reproductive technology.
The protective effects of melatonin on testis, sperm parameters quality, and in-vitro fertilization in mice following treatment with aflatoxin B1: An experimental study
Published date: Apr 26 2025
Journal Title: International Journal of Reproductive BioMedicine
Issue title: International Journal of Reproductive BioMedicine (IJRM): Volume 23, Issue No. 2
Pages: 185 – 198
Authors:
Abstract:
Background: Aflatoxin B1 (AFB1) contamination of foods and animal feeds is a public health issue. Exposure to AFB1 induces oxidative stress and can cause male reproductive toxicity. Melatonin (MLT) is a neuro-hormone produced by the pineal gland and the testis and is known as a potent antioxidant.
Objective: This study aims to determine the protective effect of MLT on testicular tissue alterations, sperm parameter indexes, and in vitro fertility assays in mice treated with AFB1. Materials and
Methods: In this experimental study, 28 adult male NMRI mice (8–10 wk old, 25–27 gr) were divided randomly into 4 groups: control, MLT (20 mg/kg/day, intraperitoneally), AFB1 (50 μg/kg/day, intraperitoneally) and MLT+AFB1. After 35 consecutive days, testis histological changes, sperm quality parameters, the rate of sperm with DNA damage, and in vitro fertilization outcomes up to the blastocyst stage were surveyed and compared between groups.
Results: Our results showed that AFB1 administration induced histological alterations in the testis and significantly decreased all the sperm parameters and in vitro fertility (fertilization and blastocyst formation rates) compared to control. Additionally, the percentages of immature sperms and sperms with DNA damage significantly (p < 0.001) increased in the AFB1-treated group. MLT treatment in the MLT+AFB1 group significantly increased testis quality and sperm parameters and improved in vitro fertilizaton rate and in vitro embryonic development.
Conclusion: These findings demonstrated that MLT can compensate for the adverse effects of AFB1 on the quality of testicular tissue, sperm parameters, sperm DNA, and in vitro fertilization outcomes.
Keywords: Melatonin, Aflatoxin B1, Sperm, Fertilization, Mice
References:
[1] Harlev A, Agarwal A, Gunes SO, Shetty A, du Plessis SS. Smoking and male infertility: An evidence-based review. World J Men‘s Health 2015; 33: 143–160.
[2] Zhou H, Wang J, Ma L, Chen L, Guo T, Zhang Y, et al. Oxidative DNA damage and multi-organ pathologies in male mice subchronically treated with aflatoxin B1. Ecotoxicol Environ Saf 2019; 186: 109697.
[3] Asadpour R, Aliyoldashi MH, Saberivand A, Hamidian Gh, Hejazi M. Ameliorative effect of selenium nanoparticles on the structure and function of testis and in vitro embryo development in Aflatoxin B1-exposed male mice. Andrologia 2020; 52: e13824.
[4] Cheng L, Qin Y, Hu X, Ren L, Zhang C, Wang X, et al. Melatonin protects in vitro matured porcine oocytes from toxicity of Aflatoxin B1. J Pineal Res 2019; 66: e12543.
[5] Yasin M, Mazdak R, Mino I. Aflatoxin B1 impairs spermatogenesis: An experimental study for crosslink between oxidative stress and mitochondria-dependent apoptosis. Environ Toxicol 2018; 33: 1204–1213.
[6] Huang W, Cao Z, Zhang J, Ji Q, Li Y. Aflatoxin B1 promotes autophagy associated with oxidative stress-related PI3K/AKT/mTOR signaling pathway in mice testis. Environ Pollut 2019; 255: 113317.
[7] Cao Z, Shao B, Xu F, Liu Y, Li Y, Zhu Y. Protective effect of selenium on aflatoxin B1-induced testicular toxicity in mice. Biol Trace Elem Res 2017; 180: 233–238.
[8] Adedara IA, Nanjappa MK, Farombi EO, Akingbemi BT. Aflatoxin B1 disrupts the androgen biosynthetic pathway in rat Leydig cells. Food Chem Toxicol 2014; 65: 252–259.
[9] Rajaura S, Chauhan P, Chandra H, Bhardwaj N. Aflatoxin B1 administration induces reactive oxygen species production and apoptosis of erythrocytes in mice. Toxicon 2023; 221: 106963.
[10] Rajaura S, Babu R, Bhardwaj N, Chauhan P, Singh A, Afzal M. Aflatoxin B1 administration causes inflammation and apoptosis in the lungs and spleen. Toxicon 2024; 238: 107581.
[11] Wu D, Wu Y, Zhang M, Lan H. Aflatoxin B1 exposure triggers inflammation and premature skin aging via ERMCS/Ca2+/ROS signaling cascade. Int Immunopharmacol 2023; 124: 110961.
[12] Omur AD, Yildirim B, Saglam YS, Comakli S, Ozkaraca M. Activity of resveratrol on the influence of aflatoxin B1 on the testes of Sprague dawley rats. Pol J Vet Sci 2019: 22; 313–320.
[13] Zhang P, Zheng Y, Lv Y, Li F, Su L, Qin Y, et al. Melatonin protects the mouse testis against heat-induced damage. Mol Hum Reprod 2020; 26: 65–79.
[14] Cui Y, Ren L, Li B, Fang J, Zhai Y, He X, et al. Melatonin relieves busulfan-induced spermatogonial stem cell apoptosis of mouse testis by inhibiting endoplasmic reticulum stress. Cell Physiol Biochem 2017; 44: 2407–2421.
[15] Du Z, Xu S, Hu S, Yang H, Zhou Z, Sidhu K, et al. Melatonin attenuates detrimental effects of diabetes on the niche of mouse spermatogonial stem cells by maintaining Leydig cells. Cell Death Dis 2018; 9: 968.
[16] Mohammadghasemi F, Jahromi SK. Melatonin ameliorates testicular damages induced by nicotine in mice. Iran J Basic Med Sci 2018; 21: 639–644.
[17] Gesing A, Karbownik-Lewinska M. Protective effects of melatonin and N-acetylserotonin on aflatoxin B1-induced lipid peroxidation in rats. Cell Biochem Funct 2008; 26: 314–319.
[18] Narayana K, Prashanthi N, Nayanatara A, Bairy LK, D’Souza UJA. An organophosphate insecticide methyl parathion (o-o-dimethyl o-4-nitrophenyl phosphorothioate) induces cytotoxic damage and tubular atrophy in the testis despite elevated testosterone level in the rat. J Toxicol Sci 2006; 31: 177–189.
[19] Karimipour M, Dibayi Z, Ahmadi A, Zirak Javanmard M, Hosseinalipour E. The protective effect of vitamin C on phenylhydrazine-induced hemolytic anemia on sperm quality and in-vitro embryo development in mice. Int J Reprod BioMed 2018; 16: 791–800.
[20] Zahmatkesh E, Najafi Gh, Nejati V. Protective effect of royal jelly on in vitro fertilization (IVF) in male mice treated with oxymetholone. Cell J 2015; 17: 569–575.
[21] Karimipour M, Zirak Javanmard M, Ahmadi A, Jafari A. Oral administration of titanium dioxide nanoparticle through ovarian tissue alterations impairs mice embryonic development. Int J Reprod BioMed 2018; 16: 397–404.
[22] Huang W, Cao Z, Yao Q, Ji Q, Zhang J, Li Y. Mitochondrial damage are involved in aflatoxin B1-induced testicular damage and spermatogenesis disorder in mice. Sci Total Environ 2020; 701: 135077.
[23] Mathuria N, Verma RJ. Curcumin ameliorates aflatoxin-induced toxicity in mice spermatozoa. Fertil Steril 2008; 90: 775–780.
[24] Huang W, Cao Z, Cui Y, Huo S, Shao B, Song M, et al. Lycopene ameliorates aflatoxin B1-induced testicular lesion by attenuating oxidative stress and mitochondrial damage with Nrf2 activation in mice. Ecotoxicol Environ Saf 2023; 256: 114846.
[25] Taheri Moghadam M, Dadfar R, Khorsandi L. The effects of ozone and melatonin on busulfan-induced testicular damage in mice. JBRA Assist Reprod 2021; 25: 176–184.
[26] Gao Y, Wu X, Zhao S, Zhang Y, Ma H, Yang Z, et al. Melatonin receptor depletion suppressed hCG-induced testosterone expression in mouse Leydig cells. Cell Mol Biol Lett 2019; 24: 1–14.
[27] Ramalho-Santos J, Varum S, Amaral S, Mota PC, Sousa AP, Amaral A. Mitochondrial functionality in reproduction: From gonads and gametes to embryos and embryonic stem cells. Hum Reprod Update 2009; 15: 553–572.
[28] Komsky-Elbaz A, Saktsier M, Roth Z. Aflatoxin B1 impairs sperm quality and fertilization competence. Toxicology 2018; 393: 42–50.
[29] Malm G, Haugen TB, Rylander L, Giwercman A. Seasonal fluctuation in the secretion of the antioxidant melatonin is not associated with alterations in sperm DNA damage. Asian J Androl 2017; 19: 52–56.
[30] Ma L, Wang J, Zhang Y. Probing the characterization of the interaction of aflatoxins B1 and G1 with calf thymus DNA in vitro. Toxins 2017; 9: 209.
[31] Feng WH, Xue KS, Tang L, Williams PL, Wang JS. Aflatoxin B1-induced developmental and DNA damage in Caenorhabditis elegans. Toxins 2016; 9: 9.
[32] Faisal K, Periasamy VS, Sahabudeen S, Radha A, Anandhi R, Akbarsha MA. Spermatotoxic effect of aflatoxin B1 in rat: Extrusion of outer dense fibres and associated axonemal microtubule doublets of sperm flagellum. Reproduction 2008; 135: 303–310.
[33] Agarwal A, Said TM. Role of sperm chromatin abnormalities and DNA damage in male infertility. Hum Reprod Update 2003; 9: 331–345.
[34] Wang Z, Teng Z, Wang Z, Song Z, Zhu P, Li N, et al. Melatonin ameliorates paclitaxel-induced mice spermatogenesis and fertility defects. J Cell Mol Med 2022; 26: 1219–1228.
[35] Soleimani Mehranjani M, Azizi M, Sadeghzadeh F. The effect of melatonin on testis histological changes and spermatogenesis indexes in mice following treatment with dexamethasone. Drug Chem Toxicol 2022; 45: 1140–1149.
[36] Zi T, Liu Y, Zhang Y, Wang Z, Wang Z, Zhan S, et al. Protective effect of melatonin on alleviating early oxidative stress induced by DOX in mice spermatogenesis and sperm quality maintaining. Reprod Biol Endocrinol 2022; 20: 105.
[37] Succu S, Berlinguer F, Pasciu V, Satta V, Leoni GG, Naitana S. Melatonin protects ram spermatozoa from cryopreservation injuries in a dose-dependent manner. J Pineal Res 2011; 50: 310–318.
[38] Guz J, Gackowski D, Foksinski M, Rozalski R, Zarakowska E, Siomek A, et al. Comparison of oxidative stress/DNA damage in semen and blood of fertile and infertile men. PloS One 2013; 8: e68490.