Keywords
Rat
Spermatocyte
Spermatogenesis
Antioxidant
Submitted : 2024-07-10
Accepted : 2024-07-25
Published : 2024-08-09
Abstract
In animals, heat stress (HS) disrupts spermatogenesis, reducing sperm quality and, in severe cases, potentially inducing the loss of male reproductive function. Melatonin confers significant resistance to oxidative stress and apoptosis; however, its specific effects on rat spermatocytes and the mechanism underlying its anti-HS effects remain inadequately explored. Therefore, this study aimed to analyze the effects of melatonin at different concentrations on sperm cell activity in heat-stressed rats. Modeling heat stress injury, sperm viability and density assay, sperm plasma membrane integrity analysis, and oxidative stress assay of testicular tissue were conducted. The results revealed that HS caused sperm cell injury. However, the intraperitoneal injection of melatonin effectively improved spermatozoa quality, and a dose of 1 mM significantly alleviated the HS-induced damage. Moreover, HS increased the levels of oxidative and endoplasmic reticulum (ES) stress in rat testicular tissues, inducing germ cell apoptosis and pathological changes. Similarly, melatonin treatment improved sperm cell viability and density, inhibited germ cell apoptosis, and reduced oxidative and ES stress levels. Overall, melatonin effectively reduced the adverse effects of HS on rat sperm cells, and an intraperitoneal injection of 1 mM (0.6966 mg) melatonin facilitated the normal production of spermatozoa. Notably, its mechanism may involve reduced ES and oxidative stress levels in testicular tissues, increased expression of the anti-apoptotic protein Bcl-2, and inhibition of germ cell apoptosis.
References
Bjorndahl L, Kirkman Brown J, other Editorial Board Members of the WHO Laboratory Manual for the Examination and Processing of Human Semen, 2022, The Sixth Edition of the WHO Laboratory Manual for the Examination and Processing of Human Semen: Ensuring Quality and Standardization in Basic Examination of Human Ejaculates. Fertil Steril, 117(2): 246–251. https://doi.org/10.1016/j.fertnstert.2021.12.012
Ghantabpour T, Nashtaei MS, Nekoonam S, et al., 2022, The Effect of Astaxanthin on Motility, Viability, Reactive Oxygen Species, Apoptosis, and Lipid Peroxidation of Human Spermatozoa During the Freezing-Thawing Process. Biopreserv Biobank, 20(4): 367–373. https://doi.org/10.1089/bio.2021.0112
Agarwal A, Majzoub A, Baskaran S, et al., 2020, Sperm DNA Fragmentation: A New Guideline for Clinicians. World J Mens Health, 38(4): 412–471. https://doi.org/10.5534/wjmh.200128
Fontoura P, Mello MD, Gallo-Sá P, et al., 2017, Leptin Improves Sperm Cryopreservation via Antioxidant Defense. J Reprod Infertil, 18(1): 172–178.
Baumber J, Ball BA, Linfor JJ, et al., 2003, Reactive Oxygen Species and Cryopreservation Promote DNA Fragmentation in Equine Spermatozoa. J Androl, 24(4): 621–628. https://doi.org/10.1002/j.1939-4640.2003.tb02714.x
Taylor K, Roberts P, Sanders K, et al., 2009, Effect of Antioxidant Supplementation of Cryopreservation Medium on Post-Thaw Integrity of Human Spermatozoa. Reprod Biomed Online, 18(2): 184–189. https://doi.org/10.1016/s1472-6483(10)60254-4
Reiter RJ, Rosales-Corral SA, Manchester LC, et al., 2013, Peripheral Reproductive Organ Health and Melatonin: Ready for Prime Time. Int J Mol Sci, 14(4): 7231–7272. https://doi.org/10.3390/ijms14047231
Bocheva G, Slominski RM, Janjetovic Z, et al., 2022, Protective Role of Melatonin and Its Metabolites in Skin Aging. Int J Mol Sci, 23(3): 1238. https://doi.org/10.3390/ijms23031238
Vasey C, McBride J, Penta K, 2021, Circadian Rhythm Dysregulation and Restoration: The Role of Melatonin. Nutrients, 13(10): 3480. https://doi.org/10.3390/nu13103480
Shin JW, 2023, Neuroprotective Effects of Melatonin in Neurodegenerative and Autoimmune Central Nervous System Diseases. Encephalitis, 3(2): 44–53. https://doi.org/10.47936/encephalitis.2022.00094
Atayik MC, Cakatay U, 2022, Melatonin-Related Signaling Pathways and Their Regulatory Effects in Aging Organisms. Biogerontology, 23(5): 529–539. https://doi.org/10.1007/s10522-022-09981-y
Alomari T, Al-Abdallat H, Hamamreh R, et al., 2024, Assessing the Antiviral Potential of Melatonin: A Comprehensive Systematic Review. Rev Med Virol, 34(1): e2499. https://doi.org/10.1002/rmv.2499
Hernandez-Ruiz J, Ruiz-Cano D, Giraldo-Acosta M, et al., 2022, Melatonin in Brassicaceae: Role in Postharvest and Interesting Phytochemicals. Molecules, 27(5): 1523. https://doi.org/10.3390/molecules27051523
Liu WX, Tan SJ, Wang YF, et al., 2022, Melatonin Promotes the Proliferation of Primordial Germ Cell-Like Cells Derived from Porcine Skin-Derived Stem Cells: A Mechanistic Analysis. J Pineal Res, 73(4): e12833. https://doi.org/10.1111/jpi.12833
Li Y, Zhang J, Wan J, et al., 2020, Melatonin Regulates ABeta Production/Clearance Balance and ABeta Neurotoxicity: A Potential Therapeutic Molecule for Alzheimer’s Disease. Biomed Pharmacother, 132: 110887. https://doi.org/10.1016/j.biopha.2020.110887
Guo YL, Wei XJ, Zhang T, et al., 2023, Molecular Mechanisms of Melatonin-Induced Alleviation of Synaptic Dysfunction and Neuroinflammation in Parkinson’s Disease: A Review. Eur Rev Med Pharmacol Sci, 27(11): 5070–5082. https://doi.org/10.26355/eurrev_202306_32624
Lang U, Rivest RW, Schlaepfer LV, et al., 1984, Diurnal Rhythm of Melatonin Action on Sexual Maturation of Male Rats. Neuroendocrinology, 38(4): 261–268. https://doi.org/10.1159/000123901
Salem M, Khadivi F, Feizollahi N, et al., 2024, Melatonin Promotes Differentiation of Human Spermatogonial Stem Cells Cultured on Three-Dimensional Decellularized Human Testis Matrix. Urol J, 21(4): 250–264. https://doi.org/10.22037/uj.v20i.7846
Deng SL, Zhang Y, Yu K, et al., 2017, Melatonin Up-Regulates the Expression of the GATA-4 Transcription Factor and Increases Testosterone Secretion from Leydig Cells through RORAlpha Signaling in An In Vitro Goat Spermatogonial Stem Cell Differentiation Culture System. Oncotarget, 8(66): 110592–110605. https://doi.org/10.18632/oncotarget.22855
Haghi-Aminjan H, Asghari MH, Farhood B, et al., 2018, The Role of Melatonin on Chemotherapy-Induced Reproductive Toxicity. J Pharm Pharmacol, 70(3): 291–306. https://doi.org/10.1111/jphp.12855
Gao Y, Wu X, Zhao S, et al., 2019, Melatonin Receptor Depletion Suppressed hCG-Induced Testosterone Expression in Mouse Leydig Cells. Cell Mol Biol Lett, 24: 21. https://doi.org/10.1186/s11658-019-0147-z
Trost N, Mbengue N, Kaessmann H, 2023, The Molecular Evolution of Mammalian Spermatogenesis. Cells Dev, 175: 203865. https://doi.org/10.1016/j.cdev.2023.203865
Yildiz C, Coskun Cetin N, Yavas I, et al., 2022, The Effect of Different Preservation Media and Temperatures on Sperm Quality and DNA Integrity in Mouse Cauda Spermatozoa. Cryo Letters, 43(6): 349–356.
Fu Y, Shen W, Bai H, et al., 2024, Roles of Y-27632 on Sheep Sperm Metabolism. J Anim Sci, 102: skae020. https://doi.org/10.1093/jas/skae020
Khan FA, Gartley CJ, Khanam A, 2018, Canine Cryptorchidism: An Update. Reprod Domest Anim, 53(6): 1263–1270. https://doi.org/10.1111/rda.13231
Gao Y, Wang C, Wang K, et al., 2022, The Effects and Molecular Mechanism of Heat Stress on Spermatogenesis and the Mitigation Measures. Syst Biol Reprod Med, 68(5–6): 331–347. https://doi.org/10.1080/19396368.2022.2074325
Shahat AM, Rizzoto G, Kastelic JP, 2020, Amelioration of Heat Stress-Induced Damage to Testes and Sperm Quality. Theriogenology, 158: 84–96. https://doi.org/10.1016/j.theriogenology.2020.08.034
Zarkovic N, 2020, Roles and Functions of ROS and RNS in Cellular Physiology and Pathology. Cells, 9(3): 767. https://doi.org/10.3390/cells9030767
Maya-Cano DA, Arango-Varela S, Santa-Gonzalez GA, 2021, Phenolic Compounds of Blueberries (Vaccinium spp) as a Protective Strategy Against Skin Cell Damage Induced by ROS: A Review of Antioxidant Potential and Antiproliferative Capacity. Heliyon, 7(2): e06297. https://doi.org/10.1016/j.heliyon.2021.e06297
Du Plessis SS, Agarwal A, Halabi J, et al., 2015, Contemporary Evidence on the Physiological Role of Reactive Oxygen Species in Human Sperm Function. J Assist Reprod Genet, 32(4): 509–520. https://doi.org/10.1007/s10815-014-0425-7
de Lamirande E, Jiang H, Zini A, et al., 1997, Reactive Oxygen Species and Sperm Physiology. Rev Reprod, 2(1): 48–54. https://doi.org/10.1530/ror.0.0020048
Zhang HM, Zhang Y, 2014, Melatonin: A Well-Documented Antioxidant with Conditional Pro-Oxidant Actions. J Pineal Res, 57(2): 131–146. https://doi.org/10.1111/jpi.12162
Ziaeipour S, Piryaei A, Aliaghaei A, et al., 2021, Chronic Scrotal Hyperthermia Induces Azoospermia and Severe Damage to Testicular Tissue in Mice. Acta Histochem, 123(4): 151712. https://doi.org/10.1016/j.acthis.2021.151712
Garcia CK, Robinson GP, Gambino BJ, et al., 2022, The Impact of Castration on Physiological Responses to Exertional Heat Stroke in Mice. PLoS One, 17(10): e0275715. https://doi.org/10.1371/journal.pone.0275715
Hoang-Thi AP, Dang-Thi AT, Phan-Van S, et al., 2022, The Impact of High Ambient Temperature on Human Sperm Parameters: A Meta-Analysis. Iran J Public Health, 51(4): 710–723. https://doi.org/10.18502/ijph.v51i4.9232
Capela L, Leites I, Romao R, et al., 2022, Impact of Heat Stress on Bovine Sperm Quality and Competence. Animals (Basel), 12(8): 975. https://doi.org/10.3390/ani12080975
Hamilton TRDS, Siqueira AFP, de Castro LS, et al., 2018, Effect of Heat Stress on Sperm DNA: Protamine Assessment in Ram Spermatozoa and Testicle. Oxid Med Cell Longev, 2018: 5413056. https://doi.org/10.1155/2018/5413056
Abdollahi F, Farhang Dehghan S, Amanpour S, et al., 2021, Effect of Co-exposure to Heat and Psychological Stressors on Sperm DNA and Semen Parameters. Toxicol Rep, 8: 1948–1954. https://doi.org/10.1016/j.toxrep.2021.11.015
Sciorio R, Tramontano L, Esteves SC, 2022, Effects of Mobile Phone Radiofrequency Radiation on Sperm Quality. Zygote, 30(2): 159–168. https://doi.org/10.1017/S096719942100037X
Kesari KK, Agarwal A, Henkel R, 2018, Radiations and Male Fertility. Reprod Biol Endocrinol, 16(1): 118. https://doi.org/10.1186/s12958-018-0431-1
Jimoh OA, Daramola OT, Okin-Aminu HO, et al., 2023, Effect of Phytogenic Supplements on the Reproductive Physiology and Metabolic Hormones of Rabbits Exposed to Heat Stress Conditions. J Therm Biol, 112: 103438. https://doi.org/10.1016/j.jtherbio.2022.103438
Abdelnour SA, Swelum AA, Sindi RA, et al., 2022, Responses of Sperm Mitochondria Functionality in Animals to Thermal Stress: The Mitigating Effects of Dietary Natural Antioxidants. Reprod Domest Anim, 57(10): 1101–1112. https://doi.org/10.1111/rda.14193
El-Gindy YM, Sabir SA, Zahran SM, et al., 2023, Effect of Dietary Onion (Allium cepa L.) Powder as An Antioxidant on Semen Quality, Blood Biochemicals, and Reproductive Parameters, as well as Immunological Variables of Rabbit Bucks Under Severe Heat Stress. Trop Anim Health Prod, 55(6): 380. https://doi.org/10.1007/s11250-023-03788-w
Shadmehr S, Fatemi Tabatabaei SR, Hosseinifar S, et al., 2018, Attenuation of Heat Stress-Induced Spermatogenesis Complications by Betaine in Mice. Theriogenology, 106: 117–126. https://doi.org/10.1016/j.theriogenology.2017.10.008
Zhang Y, Liu T, Yang H, et al., 2022, Melatonin: A Novel Candidate for the Treatment of Osteoarthritis. Ageing Res Rev, 78: 101635. https://doi.org/10.1016/j.arr.2022.101635
Bondy SC, 2023, Melatonin and Aging. Subcell Biochem, 103: 291–307. https://doi.org/10.1007/978-3-031-26576-1_13
Repova K, Baka T, Krajcirovicova K, et al., 2022, Melatonin as a Potential Approach to Anxiety Treatment. Int J Mol Sci, 23(24): 16187. https://doi.org/10.3390/ijms232416187
Zhang X, Zheng Y, Wang Z, et al., 2023, Melatonin as a Therapeutic Agent for Alleviating Endothelial Dysfunction in Cardiovascular Diseases: Emphasis on Oxidative Stress. Biomed Pharmacother, 167: 115475. https://doi.org/10.1016/j.biopha.2023.115475
Gan M, Jing Y, Xie Z, et al., 2023, Potential Function of Testicular MicroRNAs in Heat-Stress-Induced Spermatogenesis Disorders. Int J Mol Sci, 24(10): 8809. https://doi.org/10.3390/ijms24108809
Zhao F, Whiting S, Lambourne S, et al., 2021, Melatonin Alleviates Heat Stress-Induced Oxidative Stress and Apoptosis in Human Spermatozoa. Free Radic Biol Med, 164: 410–416. https://doi.org/10.1016/j.freeradbiomed.2021.01.014
Qin DZ, Cai H, He C, et al., 2021, Melatonin Relieves Heat-Induced Spermatocyte Apoptosis in Mouse Testes by Inhibition of ATF6 and PERK Signaling Pathways. Zool Res, 42(4): 514–524. https://doi.org/10.24272/j.issn.2095-8137.2021.041
Yoon YM, Kim HJ, Lee JH, et al., 2019, Melatonin Enhances Mitophagy by Upregulating Expression of Heat Shock 70 kDa Protein 1L in Human Mesenchymal Stem Cells under Oxidative Stress. Int J Mol Sci, 20(18): 4545. https://doi.org/10.3390/ijms20184545
Li C, Ren C, Chen Y, et al., 2023, Changes on Proteomic and Metabolomic Profiling of Cryopreserved Sperm Effected by Melatonin. J Proteomics, 273: 104791. https://doi.org/10.1016/j.jprot.2022.104791