Alpha-Lipoic Acid Ameliorates Sperm DNA Damage and Chromatin Integrity in Men with High DNA Damage: A Triple Blind Randomized Clinical Trial

Document Type : Original Article

Authors

1 Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran

2 Department of Animal Biotechnology, Reproductive Biomedicine Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran

3 Isfahan Fertility and Infertility Center, Isfahan, Iran

4 Department of Andrology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran

Abstract

Objective: Evidence suggests the contributory role of oxidative stress (OS) to sperm DNA damage and eventually, male infertility. Antioxidant supplementation has exhibited favorable results regarding seminal OS, sperm DNA damage, and chromatin integrity. We aimed to evaluate the effect of alpha-lipoic acid (ALA) supplementation on semen analysis, sperm DNA damage, chromatin integrity, and seminal/intracellular OS in infertile men with high sperm DNA damage.
Materials and Methods: In this randomized triple-blind placebo-controlled clinical trial study, we opted for a triple-blind controlled clinical trial design. Considering the study’s inclusion criteria for the level of sperm DNA fragmentation (higher than the threshold of 30 and 15%), 70% of participants were selected for this clinical research study. Subjects were divided into case and control groups receiving oral ALA (600 mg/day) and placebo for eighty days, respectively. Sperm parameters and functional tests were examined and compared before and after treatment. The final sample size was 34 and 29 for ALA and placebo receivers, respectively.
Results: No significant differences were observed about anthropometrics and baseline measures of semen analysis, DNA damage, OS, and chromatin integrity between the two groups. Conventional semen parameters were enhanced insignificantly in both groups (P>0.05). DNA damage decreased significantly in the ALA group, as per sperm chromatin structure assay (SCSA, P<0.001). Moreover, chromomycin A3 (CMA3) staining results indicated a decrease in nuclear protamine deficiency post-ALA therapy (P=0.004). Lipid peroxidation decreased significantly after treatment with ALA
(P=0.003). Further, seminal antioxidant capacity/activity did not differ significantly in either of the groups (registration number: IRCT20190406043177N1).
Conclusion: An 80-day course of oral ALA supplementation (600 mg/day) alleviates sperm OS, DNA damage, and chromatin integrity in men with high sperm DNA damage.

Keywords

References

  1. Zegers-Hochschild F, Adamson GD, Dyer S, Racowsky C, de Mouzon J, Sokol R, et al. The International Glossary on Infertility and Fertility Care, 2017. Fertil Steril. 2017; 108(3): 393-406.
  2. Agarwal A, Mulgund A, Hamada A, Chyatte MR. A unique view on male infertility around the globe. Reprod Biol Endocrinol. 2015; 13: 37.
  3. World Health Organization. WHO laboratory manual for the examination and processing of human semen. 6th ed. Geneva: World Health Organization; 2021.
  4. Moghissi K, Wallach E. Unexplained infertility. Fertil Steril. 1983; 39(1): 5-21.
  5. Haddock L, Gordon S, Lewis SEM, Larsen P, Shehata A, Shehata H. Sperm DNA fragmentation is a novel biomarker for early pregnancy loss. Reprod Biomed Online. 2021; 42(1): 175-184.
  6. Aitken RJ, Bakos HW. Should we be measuring DNA damage in human spermatozoa? New light on an old question. Hum Reprod. 2021; 36(5): 1175-1185.
  7. Wagner H, Cheng JW, Ko EY. Role of reactive oxygen species in male infertility: an updated review of literature. Arab J Urol. 2017; 16(1): 35-43.
  8. Pasqualotto FF, Sharma RK, Kobayashi H, Nelson DR, Thomas AJ, Agarwal A. Oxidative stress in normospermic men undergoing infertility evaluation. J Androl. 2001; 22(2): 316-322.
  9. du Plessis SS, Agarwal A, Halabi J, Tvrda E. Contemporary evidence on the physiological role of reactive oxygen species in human sperm function. J Assist Reprod Genet. 2015; 32(4): 509-520.
  10. Rashki Ghaleno L, Alizadeh A, Drevet JR, Shahverdi A, Valojerdi MR. Oxidation of sperm DNA and male infertility. Antioxidants (Basel). 2021; 10(1): 97.
  11. Gualtieri R, Kalthur G, Barbato V, Longobardi S, di Rella F, Adiga SK, et al. Sperm oxidative stress during in vitro manipulation and its effects on sperm function and embryo development. Antioxidants. 2021; 10(7): 1025.
  12. Lewis SEM, Simon L. Clinical implications of sperm DNA damage. Hum Fertil (Camb). 2010; 13(4): 201-207.
  13. Duca Y, Calogero AE, Cannarella R, Condorelli RA, la Vignera S. Current and emerging medical therapeutic agents for idiopathic male infertility. Expert Opin Pharmacother. 2019; 20(1): 55-67.
  14. Salehi B, Berkay Yılmaz Y, Antika G, Boyunegmez Tumer T, Fawzi Mahomoodally M, Lobine D, et al. Insights on the use of α-lipoicacid for therapeutic purposes. Biomolecules. 2019; 9(8): 356.
  15. Shay KP, Moreau RF, Smith EJ, Smith AR, Hagen TM. Alpha-lipoic acid as a dietary supplement: Molecular mechanisms and therapeutic potential. Biochim Biophys Acta. 2009; 1790(10): 1149-1160.
  16. Prathima P, Pavani R, Sukeerthi S, Sainath SB. α-Lipoic acid inhibits testicular and epididymal oxidative damage and improves fertility efficacy in arsenic-intoxicated rats. J Biochem Mol Toxicol. 2018; 32(2).
  17. Truong T, Gardner DK. Antioxidants improve IVF outcome and subsequent embryo development in the mouse. Hum Reprod. 2017; 32(12): 2404-2413.
  18. Haghighian HK, Haidari F, Mohammadi-Asl J, Dadfar M. Randomized, triple-blind, placebo-controlled clinical trial examining the effects of alpha-lipoic acid supplement on the spermatogram and seminal oxidative stress in infertile men. Fertil Steril. 2015; 104(2): 318-324.
  19. Abbasi B, Molavi N, Tavalaee M, Abbasi H, Nasr-Esfahani MH. Alpha-lipoic acid improves sperm motility in infertile men after varicocelectomy: a triple-blind randomized controlled trial. Reprod Biomed Online. 2020; 41(6): 1084-1091.
  20. Evenson D, Wixon R. Meta-analysis of sperm DNA fragmentation using the sperm chromatin structure assay. Reprod Biomed Online. 2006; 12(4): 466-472.
  21. Henkel R, Hajimohammad M, Stalf T, Hoogendijk C, Mehnert C, Menkveld R, et al. Influence of deoxyribonucleic acid damage on fertilization and pregnancy. Fertil Steril. 2004; 81(4): 965-972.
  22. Evenson DP. Sperm chromatin structure assay (SCSA ® ). Methods Mol Biol. 2013; 927: 147-164.
  23. Auger J, Mesbah M, Huber C, Dadoune JP. Aniline blue staining as a marker of sperm chromatin defects associated with different semen characteristics discriminates between proven fertile and suspected infertile men. Int J Androl. 1990; 13(6): 452-462.
  24. Iranpour FG, Nasr-Esfahani MH, Valojerdi MR, Taki Al-Taraihi TM. Chromomycin A3 staining as a useful tool for evaluation of male fertility. J Assist Reprod Genet. 2000; 17(1): 60-66.
  25. Sivandzade F, Bhalerao A, Cucullo L. Analysis of the mitochondrial membrane potential using the cationic JC-1 dye as a sensitive fluorescent probe. Bio Protoc. 2019; 9(1): e3128.
  26. Agarwal A, Varghese AC, Sharma RK. Markers of oxidative stress and sperm chromatin integrity. Methods Mol Biol. 2009; 590: 377- 402.
  27. Bender Atik R, Christiansen OB, Elson J, Kolte AM, Lewis S, Middeldorp S, et al. ESHRE guideline: recurrent pregnancy loss. Hum Reprod Open. 2018; 2018(2): hoy004.
  28. Agarwal A, Parekh N, Panner Selvam MK, Henkel R, Shah R, Homa ST, et al. Male oxidative stress infertility (MOSI): Proposed terminology and clinical practice guidelines for management of idiopathic male infertility. World J Mens Health. 2019; 37(3): 296-312.
  29. Arafa M, Agarwal A, Majzoub A, Panner Selvam MK, Baskaran S, Henkel R, et al. Efficacy of antioxidant supplementation on conventional and advanced sperm function tests in patients with idiopathic male infertility. Antioxidants. 2020; 9(3): 219.
  30. Hekmatdoost A, Lakpour N, Sadeghi MR. Sperm chromatin integrity: etiologies and mechanisms of abnormality, assays, clinical importance, preventing and repairing damage. Avicenna J Med Biotechnol. 2009; 1(3): 147-160.
  31. Aoki VW, Moskovtsev SI, Willis J, Liu L, Mullen JBM, Carrell DT. DNA integrity is compromised in protamine-deficient human sperm. J Androl. 2005; 26(6): 741-748.
  32. Manochantr S, Chiamchanya C, Sobhon P. Relationship between chromatin condensation, DNA integrity and quality of ejaculated spermatozoa from infertile men. Andrologia. 2012; 44(3): 187-199.
  33. Cronan JE. Biotin and lipoic acid: synthesis, attachment, and regulation. EcoSal Plus. 2014; 6(1): 10.1128/ecosalplus.ESP-0001- 2012.
  34. Henkel R, Hoogendijk CF, Bouic PJD, Kruger TF. TUNEL assay and SCSA determine different aspects of sperm DNA damage. Andrologia. 2010; 42(5): 305-313.
  35. Gallon F, Marchetti C, Jouy N, Marchetti P. The functionality of mitochondria differentiates human spermatozoa with high and low fertilizing capability. Fertil Steril. 2006; 86(5): 1526-1530.
  36. 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; 52(9): 953-960.
  37. Mahfouz RZ, du Plessis SS, Aziz N, Sharma R, Sabanegh E, Agarwal A. Sperm viability, apoptosis, and intracellular reactive oxygen species levels in human spermatozoa before and after induction of oxidative stress. Fertil Steril. 2010; 93(3): 814-821.
  38. Dutta S, Majzoub A, Agarwal A. Oxidative stress and sperm function: a systematic review on evaluation and management. Arab J Urol. 2019; 17(2): 87-97.
  39. Agarwal A, Virk G, Ong C, du Plessis SS. Effect of oxidative stress on male reproduction. World J Mens Health. 2014; 32(1): 1-17.

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