Severe deterioration in sperm parameters and testes of rats administered naproxen and diclofenac at pre-puberty: An experimental study
Background: Although naproxen and diclofenac are the most commonly used nonsteroidal drugs, their toxicity affecting male reproductivity has not been sufficiently studied, particularly when used in pre-puberty.
Objective: This study aims to investigate the toxic effects of naproxen and diclofenac on sperm parameters and testes in pre-pubertal rats.
Materials and Methods: In this experimental study, 15 pre-pubertal male albino rats (aged 5 wk, weighted 70-80 gr) were used. The animals were divided in to 3 groups (n = 5/each) of control (0.1 ml dimethyl sulfoxide), naproxen (50 mg/kg), and diclofenac sodium (5 mg/kg), and were orally administered with these drugs every day for 3 wk. Epididymal spermatozoa were taken to assess sperm count, viability, and morphology. After preparing tissue sections, testicular histopathological and histomorphometric analyses were performed.
Results: The body weights of rats in both naproxen and diclofenac groups significantly decreased (p < 0.001 and p = 0.03, respectively) in comparison to the control group. Remarkably, the testis weight and total sperm number significantly decreased (p = 0.002, and p = 0.004 respectively) in the naproxen-administered rats only. The sperm viability percentage decreased significantly in both diclofenac and naproxenadministered groups (p ≤ 0.001 and p = 0.03 respectively). Moreover, there was a significant increase (p < 0.001) in the percentage of sperm morphological anomalies in both drug-administered groups. Also, the histological and morphometric findings exhibited severe histopathological appearances in the testes and seminiferous tubules parameters in both drug-administered groups.
Conclusion: Naproxen and diclofenac administrations of rats before their puberty induce considerable harm to sperm parameters and testicular histology and morphometry. These severe toxic effects can lead to potential infertility.
Key words: Male infertility, Nonsteroidal drugs, Pathology, Spermatozoa, Testis.
 Griswold MD. Spermatogenesis: The commitment to meiosis. Physiol Rev 2016; 96: 1–17.
 Ding J, Shang X, Zhang Zh, Jing H, Shao J, Fei Q, et al. FDA-approved medications that impair human spermatogenesis. Oncotarget 2017; 8: 10714–10725.
 Wallace JL. Mechanisms, prevention and clinical implications of nonsteroidal anti-inflammatory drugenteropathy. World J Gastroenterol 2013; 19: 1861– 1876.
 Cooper C, Chapurlat R, Al-Daghri N, Herrero-Beaumont G, Bruyere O, Rannou F, et al. Safety of oral non-selective non-steroidal anti-inflammatory drugs in osteoarthritis: What does the literature say? Drugs Aging 2019; 36: 15–24.
 Trappe TA, Liu SZ. Effects of prostaglandins and COXinhibiting drugs on skeletal muscle adaptations to exercise. J Appl Physiol 2013; 115: 909–919.
 Pier B, Edmonds JW, Wilson L, Arabshahi A, Moore R, Bates GW, et al. Comprehensive profiling of prostaglandins in human ovarian follicular fluid using mass spectrometry. Prostaglandins Other Lipid Mediat 2018; 134: 7–15.
 Niringiyumukiza JD, Cai H, Xiang W. Prostaglandin E2 involvement in mammalian female fertility: Ovulation, fertilization, embryo development and early implantation. Reprod Biol Endocrinol 2018; 16: 43.
 Duffy DM. Novel contraceptive targets to inhibit ovulation: The prostaglandin E2 pathway. Hum Reprod Update 2015; 21: 652–670.
 Frungieri MB, Calandra RS, Mayerhofer A, Matzkin ME. Cyclooxygenase and prostaglandins in somatic cell populations of the testis. Reproduction 2015; 149: R169–180.
 Kaewmala K, Uddin MJ, Cinar MU, Große-Brinkhaus C, Jonas E, Tesfaye D, et al. Investigation into association and expression of PLCz and COX-2 as candidate genes for boar sperm quality and fertility. Reprod Domest Anim 2012; 47: 213–223.
 Campos Silva R, Coelho Britto DM, de Fátima Pereira W, Brito-Melo GEA, Machado CT, Pedreira MM. Effect of short- and medium-term toxicity of doxorubicin on spermatogenesis in adult Wistar rats. Reprod Biol 2018; 18: 169–176.
 Momeni HR, Eskandari N. Effect of vitamin E on sperm parameters and DNA integrity in sodium arsenitetreated rats. Iran J Reprod Med 2012; 10: 249–256.
 Uzun B, Atli O, Perk BO, Burukoglu D, Ilgin S. Evaluation of the reproductive toxicity of naproxen sodium and meloxicam in male rats. Hum Exp Toxicol 2015; 34: 415–429.
 Mousa AA, Elweza AE, Elbaz HT, Tahoun EAE-A, Shoghy KM, Elsayed I, et al. Eucalyptus Globulus protects against diclofenac sodium induced hepatorenal and testicular toxicity in male rats. J Tradit Complement Med 2020; 10: 521–528.
 Adeyemi WJ, Omoniyi JA, Olayiwola A, Ibrahim M, Ogunyemi O, Olayaki LA. Elevated reproductive toxicity effects of diclofenac after withdrawal: Investigation of the therapeutic role of melatonin. Toxicol Rep 2019; 6: 571–577.
 Vyas A, Purohit A, Ram H. Assessment of dosedependent reproductive toxicity of diclofenac sodium in male rats. Drug Chem Toxicol 2019; 42: 478–486.
 Oumaima A, Tesnim A, Zohra H, Amira S, Ines Z, Sana C, et al. Investigation on the origin of sperm morphological defects: Oxidative attacks, chromatin immaturity, and DNA fragmentation. Environ Sci Pollut Res Int 2018; 25: 13775–13786.
 Jakubik-Uljasz J, Gill K, Rosiak-Gill A, Piasecka M. Relationship between sperm morphology and sperm DNA dispersion. Transl Androl Urol 2020; 9: 405–415.
 Ammar O, Mehdi M, Muratori M. Teratozoospermia: Its association with sperm DNA defects, apoptotic alterations, and oxidative stress. Andrology 2020; 8: 1095–1106.
 Shukla KK, Mahdi AA, Rajender S. Apoptosis, spermatogenesis and male infertility. Front Biosci 2012; 4: 746–754.
 Yousefalizadegan N, Mousavi Z, Rastegar T, Razavi Y, Najafizadeh P. Reproductive toxicity of manganese dioxide in forms of micro- and nanoparticles in male rats. Int J Reprod BioMed 2019; 17: 361–370.
 Moridian M, Khorsandi L, Talebi AR. Morphometric and stereological assessment of the effects of zinc oxide nanoparticles on the mouse testicular tissue. Bratisl Lek Listy 2015; 116: 321–325.
 Abarikwu SO, Simple G, Onuoha CS. Morphometric evaluation of the seminiferous tubules and the antioxidant protective effects of gallic acid and quercetin in the testis and liver of Butyl Phthalate treated rats. Indian J Clin Biochem 2020; 35: 20–31.
 Tripathi UK, Chhillar S, Kumaresan A, Aslam MKM, Rajak SK, Nayak S, et al. Morphometric evaluation of seminiferous tubule and proportionate numerical analysis of Sertoli and spermatogenic cells indicate differences between crossbred and purebred bulls. Vet World 2015; 8: 645–650.
 Ma L, Guo Y, Yuan Y, Li Y-G, Deng X-Z, Yang Z-W. Morphometric study of the testis and reproductive tract (including sperm granuloma) after vasectomy in mature rats. Asian J Androl 2016; 18: 66–73.
 Motawi TMK, Bustanji Y, El-Maraghy SA, Taha MO, Al Ghussein MAS. Naproxen and cromolyn as new glycogen synthase kinase 3β inhibitors for amelioration of diabetes and obesity: An investigation by docking simulation and subsequent in vitro/in vivo biochemical evaluation. J Biochem Mol Toxicol 2013; 27: 425–436.
 Villarroel-Espindola F, Maldonado R, Mancilla H, vander Stelt K, Acuna AI, Covarrubias A, et al. Muscle glycogen synthase isoform is responsible for testicular glycogen synthesis: Glycogen overproduction induces apoptosis in male germ cells. J Cell Biochem 2013; 114: 1653–1664.
 Vyas A, Ram H, Purohit A, Jatwa R. Adverse effects of subchronic dose of aspirin on reproductive profile of male rats. J Pharm 2016; 2016: 6585430.