Analyze Diet
Antioxidants (Basel, Switzerland)2024; 13(3); 322; doi: 10.3390/antiox13030322

Impact of Seminal Plasma Antioxidants on DNA Fragmentation and Lipid Peroxidation of Frozen-Thawed Horse Sperm.

Abstract: Cryopreservation is a stressful process for sperm, as it is associated with an increased production of reactive oxygen species (ROS). Elevated ROS levels, which create an imbalance with antioxidant capacity, may result in membrane lipid peroxidation (LPO), protein damage and DNA fragmentation. This study aimed to determine whether the membrane LPO and DNA fragmentation of frozen-thawed horse sperm relies upon antioxidant activity, including enzymes (superoxide dismutase (SOD), glutathione peroxidase (GPX), catalase (CAT) and paraoxonase type 1 (PON1)); non-enzymatic antioxidant capacity (Trolox-equivalent antioxidant capacity (TEAC), plasma ferric reducing antioxidant capacity (FRAP) and cupric reducing antioxidant capacity (CUPRAC)); and the oxidative stress index (OSI) of their seminal plasma (SP). Based on total motility and plasma membrane integrity (SYBR14/PI) after thawing, ejaculates were hierarchically ( < 0.001) clustered into two groups of good- (GFEs) and poor-(PFEs) freezability ejaculates. LPO and DNA fragmentation (global DNA breaks) were higher ( < 0.05) in the PFE group than in the GFE group, with LPO and DNA fragmentation (global DNA breaks) after thawing showing a positive relationship ( < 0.05) with SP OSI levels and ROS production. In addition, sperm motility and membrane integrity after thawing were negatively ( < 0.05) correlated with the activity levels of SP antioxidants (PON1 and TEAC). The present results indicate that LPO and DNA fragmentation in frozen-thawed horse sperm vary between ejaculates. These differences could result from variations in the activity of antioxidants (PON1 and TEAC) and the balance between the oxidant and antioxidant components present in the SP.
Publication Date: 2024-03-06 PubMed ID: 38539855DOI: 10.3390/antiox13030322Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research article investigates how the antioxidant activity in horse sperm affects the sperm’s DNA fragmentation and lipid peroxidation during the freezing and thawing process. The focus is on the role of several varying types of antioxidants in this process.

Study Objective

  • The main aim of this study was to ascertain if the extent of DNA fragmentation and lipid peroxidation, a process that damages sperm cells, in frozen-thawed horse sperm depends on the antioxidant activity in their seminal fluid.

Specific Parameters Analyzed

  • The study examined several factors in the seminal plasma (SP), including enzymatic antioxidants like superoxide dismutase (SOD), glutathione peroxidase (GPX), catalase (CAT), and paraoxonase type 1 (PON1).
  • The research also evaluated non-enzymatic antioxidant capability through Trolox-equivalent antioxidant capacity (TEAC), plasma ferric reducing antioxidant capacity (FRAP), and cupric reducing antioxidant capacity (CUPRAC).
  • The oxidative stress index (OSI) of the seminal plasma was also considered.

Methodology and Classification

  • The ejaculates were ranked into groups classified by their ‘freezability’. This categorization was based on total sperm motility and plasma membrane integrity after the freezing-thawing process.
  • The two groups were good-freezability ejaculates (GFEs) and poor-freezability ejaculates (PFEs).

Findings

  • The study found higher levels of lipid peroxidation and DNA fragmentation in the poor-freezability group compared to the good-freezability group.
  • Notably, these levels showed a positive correlation with the seminal plasma oxidative stress index and ROS production.
  • Furthermore, sperm motility and membrane integrity after thawing showed a negative correlation with the antioxidant activity in the seminal plasma, particularly in the case of PON1 and TEAC.

Conclusions

  • The research concludes that the extent of lipid peroxidation and DNA fragmentation in frozen-thawed horse sperm varies between ejaculates.
  • The variation could be caused by the difference in activity of the antioxidants PON1 and TEAC, and the balance between oxidant and antioxidant components present in the seminal plasma.

Cite This Article

APA
Catalán J, Yánez-Ortiz I, Torres-Garrido M, Ribas-Maynou J, Llavanera M, Barranco I, Yeste M, Miró J. (2024). Impact of Seminal Plasma Antioxidants on DNA Fragmentation and Lipid Peroxidation of Frozen-Thawed Horse Sperm. Antioxidants (Basel), 13(3), 322. https://doi.org/10.3390/antiox13030322

Publication

ISSN: 2076-3921
NlmUniqueID: 101668981
Country: Switzerland
Language: English
Volume: 13
Issue: 3
PII: 322

Researcher Affiliations

Catalán, Jaime
  • Biotechnology of Animal and Human Reproduction (TechnoSperm), Institute of Food and Agricultural Technology, University of Girona, ES-17003 Girona, Spain.
  • Unit of Cell Biology, Department of Biology, Faculty of Sciences, University of Girona, ES-17003 Girona, Spain.
  • Equine Reproduction Service, Department of Animal Medicine and Surgery, Faculty of Veterinary Sciences, Autonomous University of Barcelona, ES-08193 Cerdanyola del Vallès, Spain.
Yánez-Ortiz, Iván
  • Biotechnology of Animal and Human Reproduction (TechnoSperm), Institute of Food and Agricultural Technology, University of Girona, ES-17003 Girona, Spain.
  • Unit of Cell Biology, Department of Biology, Faculty of Sciences, University of Girona, ES-17003 Girona, Spain.
  • Equine Reproduction Service, Department of Animal Medicine and Surgery, Faculty of Veterinary Sciences, Autonomous University of Barcelona, ES-08193 Cerdanyola del Vallès, Spain.
  • School of Veterinary Medicine, Faculty of Medical, Health and Life Sciences, International University of Ecuador, Quito 170411, Ecuador.
Torres-Garrido, Marc
  • Biotechnology of Animal and Human Reproduction (TechnoSperm), Institute of Food and Agricultural Technology, University of Girona, ES-17003 Girona, Spain.
  • Unit of Cell Biology, Department of Biology, Faculty of Sciences, University of Girona, ES-17003 Girona, Spain.
Ribas-Maynou, Jordi
  • Biotechnology of Animal and Human Reproduction (TechnoSperm), Institute of Food and Agricultural Technology, University of Girona, ES-17003 Girona, Spain.
  • Unit of Cell Biology, Department of Biology, Faculty of Sciences, University of Girona, ES-17003 Girona, Spain.
Llavanera, Marc
  • Biotechnology of Animal and Human Reproduction (TechnoSperm), Institute of Food and Agricultural Technology, University of Girona, ES-17003 Girona, Spain.
  • Unit of Cell Biology, Department of Biology, Faculty of Sciences, University of Girona, ES-17003 Girona, Spain.
Barranco, Isabel
  • Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, University of Murcia, ES-30100 Murcia, Spain.
Yeste, Marc
  • Biotechnology of Animal and Human Reproduction (TechnoSperm), Institute of Food and Agricultural Technology, University of Girona, ES-17003 Girona, Spain.
  • Unit of Cell Biology, Department of Biology, Faculty of Sciences, University of Girona, ES-17003 Girona, Spain.
  • Catalan Institution for Research and Advanced Studies (ICREA), ES-08010 Barcelona, Spain.
Miró, Jordi
  • Equine Reproduction Service, Department of Animal Medicine and Surgery, Faculty of Veterinary Sciences, Autonomous University of Barcelona, ES-08193 Cerdanyola del Vallès, Spain.

Citations

This article has been cited 12 times.
  1. Sengupta P, Dutta S, Jallo MK, Rosas IM, Roychoudhury S. Seminal Plasma and Extracellular Vesicles as Molecular Gatekeepers: Oxidative Stress, Endocrine Crosstalk, and Biomarker Discovery in Male Infertility. Curr Issues Mol Biol 2026 Jan 22;48(1).
    doi: 10.3390/cimb48010117pubmed: 41614947google scholar: lookup
  2. Dabiri M, Goss DM, Ramasamy R, Bhakat M, Gardner DK, de Graaf SP, Ebrahimi Warkiani M. Sperm DNA fragmentation and its influence on mammalian reproduction. Nat Rev Urol 2026 Jan 28;.
    doi: 10.1038/s41585-025-01123-6pubmed: 41606137google scholar: lookup
  3. Strassner FM, Demattio L, Siuda M, Malama E, Muffels G, Bollwein H. Relationships Between Metabolism of Cryopreserved Equine Sperm Determined by the Seahorse Analyzer and Sperm Characteristics Measured by Flow Cytometry and Computer-Assisted Analysis of Motility. Vet Sci 2025 Nov 21;12(12).
    doi: 10.3390/vetsci12121109pubmed: 41472089google scholar: lookup
  4. Wang S, Shi L, Zhang Z, Liu J, Xing J, Yang J, Duan J, Li B, Cao G. NT5C1B Improves Fertility of Boar Spermatozoa by Enhancing Quality and Cryotolerance During Cryopreservation. Animals (Basel) 2025 Dec 8;15(24).
    doi: 10.3390/ani15243530pubmed: 41463814google scholar: lookup
  5. Dena SM, Adeleye AO, Mohlala K, Langa BC, Opuwari CS. The Impact of Diabetes Mellitus-Related Oxidative Stress on Male Fertility: A Review. J Diabetes 2025 Oct;17(10):e70157.
    doi: 10.1111/1753-0407.70157pubmed: 41123473google scholar: lookup
  6. Boni R, Ruggiero R, De Luca F, Serritella ML, Di Palma T, Cecchini Gualandi S. Repeatability of Selected Parameters Related to Stallion Sperm Quality and Cryotolerance. Animals (Basel) 2025 Sep 26;15(19).
    doi: 10.3390/ani15192805pubmed: 41096400google scholar: lookup
  7. Ullah A, Chen W, Shi L, Wang M, Geng M, Na J, Akhtar MF, Khan MZ, Wang C. Challenges and Enhancing Strategies of Equine Semen Preservation: Nutritional and Genetic Perspectives. Vet Sci 2025 Aug 25;12(9).
    doi: 10.3390/vetsci12090807pubmed: 41012733google scholar: lookup
  8. Rong J, Leng X, Jiang K, Tan J, Dong M. Systemic impacts of diabetes on spermatogenesis and intervention strategies: multilayered mechanism analysis and cutting-edge therapeutic approaches. Reprod Biol Endocrinol 2025 Sep 24;23(1):122.
    doi: 10.1186/s12958-025-01454-4pubmed: 40993627google scholar: lookup
  9. Janati S, Behmanesh MA, Biabani E, Poormoosavi SM. Investigating the Relationship Between Semen Paraoxonase-3 Concentration, Sperm Parameters, and DNA Fragmentation in Fresh or Post-Thaw Semen of Normozoospermic Men. J Reprod Infertil 2025 Jan-Mar;26(1):13-18.
    doi: 10.18502/jri.v26i1.18777pubmed: 40757043google scholar: lookup
  10. Al-Kass Z, Morrell JM, Ntallaris T. Effect of Centrifugation of Stallion Semen Through a Low Density Colloid Prior to Freezing on Sperm Cryosurvival. Animals (Basel) 2025 Jun 25;15(13).
    doi: 10.3390/ani15131881pubmed: 40646780google scholar: lookup
  11. Onochie C, Evi K, O'Flaherty C. Role of Redox-Induced Protein Modifications in Spermatozoa in Health and Disease. Antioxidants (Basel) 2025 Jun 12;14(6).
    doi: 10.3390/antiox14060720pubmed: 40563353google scholar: lookup
  12. Catalán J, Padilla L, Maside C, Martínez-Hernández J, Yánez-Ortiz I, Tvarijonaviciute A, Barranco I, Bonet S, Miró J, Yeste M. Redox profiling of preovulatory follicular fluid in the donkey is species-specific, and contributes to modulate sperm function. Sci Rep 2025 Feb 23;15(1):6522.
    doi: 10.1038/s41598-025-91422-0pubmed: 39988588google scholar: lookup