OBJECTIVE: The aim of this study was to analyse the long-term radiation effects on human sperm. METHODS: In total, 104 samples of male donors from 2 regions of Ukraine were tested. Group 1 consisted of 32 donors from the Ivano-Frankivsk region, group 2 included 72 volunteers from the Zhytomyr region. The average age of donors in both groups was 35 ± 6 years (range 24-49). To assess the level of apoptosis, membrane mitochondrial potential, concentration of reactive oxygen species, and ploidy of sperm, flow cytometry was performed. RESULTS: The individual equivalent dose of group 1 was < 0.4 mSv and of group 2 ≥ 0.4 mSv. Live spermatozoa with signs of apoptosis were significantly higher (p < 0.05) in group 2 in comparison to group 1 (15.6% and 11.2%, respectively). Spermatozoa without violating integrity were 73.2% in group 1 and approximately 16% higher than the indices of group 2. The percentage of dead necrotic spermatozoa was twice as high in men with a predicted equivalent dose of ≥ 0.4 mSv than in comparison group. A higher percentage of spermatozoa with low mitochondrial membrane potential, di- and tetraploid was found in group 2. CONCLUSIONS: An equivalent individual dose of ≥ 0.4 mSv can cause a decrease in mitochondrial potential, an increase in the production of spermatozoa with pathological ploidy, as well as to provoke increasing apoptosis in cells.

Department of Obstetrics and Gynaecology Bukovinian State Medical University Chernivtsi Ukraine

Department of Obstetrics and Gynaecology Faculty of Medicine Pavol Jozef Safarik University in Kosice Kosice Slovak Republic

Zobrazit více v PubMed

Neto FT, Bach PV, Najari BB, Li PS, Goldstein M. Spermatogenesis in humans and its affecting factors. Semin Cell Dev Biol. 2016 Nov;59:10-26. doi: 10.1016/j.semcdb.2016.04.009. Epub 2016 Apr 30. PMID: 27143445. PubMed DOI

Rombaut C, Mertes H, Heindryckx B, Goossens E. Human in vitro spermatogenesis from pluripotent stem cells: in need of a stepwise differentiation protocol? Mol Hum Reprod. 2018 Feb 1;24(2):47-54. doi: 10.1093/molehr/gax065. PMID: 29244172. PubMed DOI

Cheburakov IuIu, Cheburakova OP. Narusheniia spermatogeneza u lits, uchastvovavshikh v likvidatsii posledstviĭ avarii na Chernobylśkoĭ AES [Disorders of spermatogenesis in people working at the clean-up of the Chernobyl nuclear power plant accident]. Radiats Biol Radioecol. 1993 Nov-Dec;33(6):771-4. Russian. PMID: 8293101. PubMed

Fischbein A, Zabludovsky N, Eltes F, Grischenko V, Bartoov B. Ultramorphological sperm characteristics in the risk assessment of health effects after radiation exposure among salvage workers in Chernobyl. Environ Health Perspect. 1997 Dec;105 Suppl 6(Suppl 6):1445-9. doi: 10.1289/ehp.97105s61445. PMID: 9467060; PMCID: PMC1469951. PubMed DOI

Kesari KK, Agarwal A, Henkel R. Radiations and male fertility. Reprod Biol Endocrinol. 2018 Dec 9;16(1):118. doi: 10.1186/s12958-018-0431-1. PMID: 30445985; PMCID: PMC6240172. PubMed DOI

Kumar D, Salian SR, Kalthur G, Uppangala S, Kumari S, Challapalli S, et al. Association between sperm DNA integrity and seminal plasma antioxidant levels in health workers occupationally exposed to ionizing radiation. Environ Res. 2014 Jul;132:297-304. doi: 10.1016/j.envres.2014.04.023. Epub 2014 May 14. PMID: 24834825. PubMed DOI

Clifton DK, Bremner WJ. The effect of testicular x-irradiation on spermatogenesis in man. A comparison with the mouse. J Androl. 1983 Nov-Dec;4(6):387-92. doi: 10.1002/j.1939-4640.1983.tb00765.x. PMID: 6654753. PubMed DOI

Howell SJ, Shalet SM. Spermatogenesis after cancer treatment: damage and recovery. J Natl Cancer Inst Monogr. 2005;(34):12-7. doi: 10.1093/jncimonographs/lgi003. PMID: 15784814. PubMed DOI

Azzam EI, Jay-Gerin JP, Pain D. Ionizing radiation-induced metabolic oxidative stress and prolonged cell injury. Cancer Lett. 2012 Dec 31;327(1-2):48-60. doi: 10.1016/j.canlet.2011.12.012. Epub 2011 Dec 17. PMID: 22182453; PMCID: PMC3980444. PubMed DOI

Halliwell B. Free radicals and antioxidants - quo vadis? Trends Pharmacol Sci. 2011 Mar;32(3):125-30. doi: 10.1016/j.tips.2010.12.002. Epub 2011 Jan 6. PMID: 21216018. PubMed DOI

Ahmad IM, Temme JB, Abdalla MY, Zimmerman MC. Redox status in workers occupationally exposed to long-term low levels of ionizing radiation: A pilot study. Redox Rep. 2016 May;21(3):139-45. doi: 10.1080/13510002.2015.1101891. Epub 2016 Feb 5. PMID: 26817988; PMCID: PMC5011313. PubMed DOI

Aitken RJ. Reactive oxygen species as mediators of sperm capacitation and pathological damage. Mol Reprod Dev. 2017 Oct;84(10):1039-1052. doi: 10.1002/mrd.22871. Epub 2017 Sep 5. PMID: 28749007. PubMed DOI

Likhtariov I.A. Dosimetry and radiation hygiene / I.A. Likhtariov, B.G. Bebeshko // Bulletin of Research Centre for Radiation Medicine AMS of Ukraine. - 2005. - Vol. 5. - P. 5-10.

Malko M.V. Assessment of the Chernobyl radiological consequences / M.V. Malko; [In the report: Research activities about radiological consequences of the Chernobyl NPS accident and social activities to assist the sufferers by the accident. Edited by Imanaka T. Research Reactor Institute]. - Kyoto University, Japan. KURRI-KR-21. ISSN 1342-0852. - 1998. - P. 65-90.

Kenigsberg J.E. Radiation effects on the population of Belarus after the Chernobyl accident and the prediction of stochastic effects / J.E. Kenigsberg, V.F. Minenko, E.E. Buglova // World Health Statistics Quarterly. - 1996. - Vol. 49(1). - P. 58-61.

Glander HJ, Schaller J. Binding of annexin V to plasma membranes of human spermatozoa: a rapid assay for detection of membrane changes after cryostorage. Mol Hum Reprod. 1999 Feb;5(2):109-15. doi: 10.1093/molehr/5.2.109. PMID: 10065865. PubMed DOI

Vermes I, Haanen C, Steffens-Nakken H, Reutelingsperger C. A novel assay for apoptosis. Flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled Annexin V. J Immunol Methods. 1995 Jul 17;184(1):39-51. doi: 10.1016/0022-1759(95)00072-i. PMID: 7622868. PubMed DOI

Zou T, Liu X, Ding S, Xing J. Evaluation of sperm mitochondrial function using rh123/PI dual fluorescent staining in asthenospermia and oligoasthenozoospermia. J Biomed Res. 2010 Sep;24(5):404-10. doi: 10.1016/S1674-8301(10)60054-1. PMID: 23554656; PMCID: PMC3596687. PubMed DOI

Evenson DP, Darzynkiewicz Z, Melamed MR. Simultaneous measurement by flow cytometry of sperm cell viability and mitochondrial membrane potential related to cell motility. J Histochem Cytochem. 1982 Mar;30(3):279-80. doi: 10.1177/30.3.6174566. PMID: 6174566. PubMed DOI

Zielonka J, Vasquez-Vivar J, Kalyanaraman B. Detection of 2-hydroxyethidium in cellular systems: a unique marker product of superoxide and hydroethidine. Nat Protoc. 2008;3(1):8-21. doi: 10.1038/nprot.2007.473. PMID: 18193017. PubMed DOI

De Iuliis GN, Wingate JK, Koppers AJ, McLaughlin EA, Aitken RJ. Definitive evidence for the nonmitochondrial production of superoxide anion by human spermatozoa. J Clin Endocrinol Metab. 2006 May;91(5):1968-75. doi: 10.1210/jc.2005-2711. Epub 2006 Feb 28. PMID: 16507629. PubMed DOI

Levek-Motola N, Soffer Y, Shochat L, Raziel A, Lewin LM, Golan R. Flow cytometry of human semen: a preliminary study of a non-invasive method for the detection of spermatogenetic defects. Hum Reprod. 2005 Dec;20(12):3469-75. doi: 10.1093/humrep/dei247. Epub 2005 Aug 25. PMID: 16123093. PubMed DOI

Lazaros L, Kaponis A, Vartholomatos G, Hatzi E, Botsari S, Plachouras N, et al. Using semen flow cytometry to evaluate association of ploidy status and chromatin condensation of spermatozoa with conventional semen parameters: clinical application in intrauterine insemination. Fertil Steril. 2011 Jan;95(1):110-5. doi: 10.1016/j.fertnstert.2010.05.012. Epub 2010 Jun 18. PMID: 20667800. PubMed DOI

Agnihotri SK, Agrawal AK, Hakim BA, Vishwakarma AL, Narender T, Sachan R, et al. Mitochondrial membrane potential (MMP) regulates sperm motility. In Vitro Cell Dev Biol Anim. 2016 Oct;52(9):953-960. doi: 10.1007/s11626-016-0061-x. Epub 2016 Jun 23. PMID: 27338736. PubMed DOI

Nugent SM, Mothersill CE, Seymour C, McClean B, Lyng FM, Murphy JE. Increased mitochondrial mass in cells with functionally compromised mitochondria after exposure to both direct gamma radiation and bystander factors. Radiat Res. 2007 Jul;168(1):134-42. doi: 10.1667/RR0769.1. PMID: 17722997. PubMed DOI