SYNERGISTIC INHIBITION OF HUMAN SPERM FUNCTION BY FUCOIDAN AND OUABAIN REVEALS A POTENT NON‑HORMONAL CONTRACEPTIVE STRATEGY
Abstract
The development of safe and effective non‑hormonal contraceptive options is a global priority, particularly given the mucosal irritation and inflammatory responses associated with chemical spermicides such as Nonoxinol‑9 and Sodium Lauryl Sulfate. Natural bioactive compounds have emerged as promising alternatives due to their diverse biological properties and potentially improved safety profiles. This study investigated the in vitro effects of Fucoidan, a sulphated polysaccharide from brown algae, and Ouabain, a cardiotonic glycoside that selectively inhibits the sperm‑specific Na⁺/K⁺‑ATPase α4 isoform, on human sperm motility and vitality. Semen samples from healthy volunteers were exposed to saline (negative control), Sodium Lauryl Sulfate (positive control), Fucoidan, Ouabain, or a Fucoidan + Ouabain combination, and evaluated at 0 and 15 minutes following WHO (2021) guidelines. All treatments reduced motility and vitality compared with the negative control. Fucoidan alone produced modest reductions, whereas Ouabain induced a more pronounced inhibitory effect. The combined treatment resulted in the greatest decline, reducing progressive motility and vitality by approximately 28% at both time points, indicating a synergistic interaction. Statistical analyses (Kruskal–Wallis and Wilcoxon tests) confirmed significant differences between treatments (p < 0.05). These findings support the potential of Fucoidan and Ouabain, particularly in combination, as candidates for the development of natural, non‑hormonal contraceptive formulations. Further studies are required to assess cytotoxicity, mucosal safety, and long‑term applicability.
Author Biographies
Graduated in Nursing and Obstetrics from the Federal University of Espírito Santo (UFES); Master's degree in Biotechnology (UFES), Postgraduate specialization in Women’s Health.
Graduated in Nursing and Obstetrics from the Federal University of Espírito Santo (UFES); PhD candidate in Public Health (UFES); Master's degree in Public Health (UFES).
Graduated in Pharmacy from the Federal University of Espírito Santo (UFES); Master's student in Biotechnology in Health (UFES).
Bachelor and Licensed in Biological Sciences from the Human and Molecular Genetics Center (NGHM) of the Department of Biological Sciences at the Federal University of Espírito Santo (UFES); Masters in Biotechnology (UFES). PhD student in Biotechnology (RENORBIO/UFES).
Graduated in Biological Sciences from the Federal University of Espírito Santo (UFES); PhD in Sciences/Genetics from the University of São Paulo (USP).Teacher, advisor and researcher in the Postgraduate Program in Biotechnology/UFES.
Postdoctoral degree in Biochemistry and Molecular Genetics (UAB) and postdoctoral degree in Cancer Molecular Genetics. Full Professor (UFES), supervisor of master's and doctoral students in Biotechnology. Coordinator of the Postgraduate Program in Biotechnology at UFES.
PhD in Physiological Sciences (UFES). Associate Professor IV of the Morphology Department of UFES; Coordinator of the doctorate of the Northeast Network of Biotechnology (RENORBIO) and advisor of master's degree/doctorate in Biotechnology (UFES).
Postdoctoral degree in Biosciences (UERJ). PhD in Nuclear Biosciences with emphasis on Molecular Oncology (UERJ/INCA). Master's degree in Pharmaceutical Sciences (UFRJ) and specialization in Biochemistry (UFES). Pharmacy degree (UFES). Professor and researcher in the Postgraduate Program in Biotechnology (UFES).
References
ABE, K. et al. Molecular structure of the Na+,K+-ATPase α4β1 isoform in its ouabain-bound conformation. International Journal of Molecular Sciences, v. 25, n. 22, p. 12397, 2024. DOI: https://doi.org/10.3390/ijms252212397
AMATYA, B. et al. SNX19 interacts with caveolin-1 and flotillin-1 to regulate D1R endocytosis and signaling. Biomedicines, v. 13, n. 2, p. 481, 2025. DOI: https://doi.org/10.3390/biomedicines13020481
BARTON, B. et al. Serine protease inhibitor disrupts sperm motility leading to reduced fertility in female mice. Biology of Reproduction, v. 103, p. 400–410, 2020. DOI: https://doi.org/10.1093/biolre/ioaa049
BENJAMINI, Y.; YEKUTIELI, D. The control of the false discovery rate in multiple testing under dependency. The Annals of Statistics, v. 29, n. 4, p. 1165–1188, 2001. DOI: https://doi.org/10.1214/aos/1013699998
CHIANESE, R.; PIERANTONI, R. Mitochondrial reactive oxygen species (ROS) production alters sperm quality. Antioxidants, v. 10, n. 1, p. 92, 2021. DOI: https://doi.org/10.3390/antiox10010092
CHOLLET, L. et al. Fucoidans in nanomedicine. Marine Drugs, v. 14, n. 8, p. 145, 2016. DOI: https://doi.org/10.3390/md14080145
CONTRERAS, R. et al. Na+/K+-ATPase: More than an electrogenic pump. International Journal of Molecular Sciences, v. 25, n. 11, p. 6122, 2024. DOI: https://doi.org/10.3390/ijms25116122
HAINEAULT, C.; LECLERC, P.; GAGNON, C. Characterization of human sperm surface antigens involved in gamete interaction. Molecular Human Reproduction, v. 9, n. 5, p. 279–286, 2003. DOI: https://doi.org/10.1093/molehr/gag037
HASSAN, M. et al. Reversibility of the reproductive toxicity of gossypol in peripubertal bulls. Theriogenology, v. 61, n. 6, p. 1171–1179, 2004. DOI: https://doi.org/10.1016/j.theriogenology.2003.07.007
HATCHER, R. A. Contraceptive technology. 21. ed. Managing Contraception, LLC, 2018.
HE, Y. et al. Anti-hyperlipidemic effect of fucoidan fractions prepared from Iceland brown algae Ascophyllum nodosum in a hyperlipidemic mice model. Marine Drugs, v. 21, 2023. DOI: https://doi.org/10.3390/md21090468
HEREDERO-JIMÉNEZ, S. et al. Caveolin-1 regulates parathyroid hormone-related protein actions on PTH receptor type 1 in bone cells. Journal of Cellular Physiology, v. 240, n. 1, 2025. DOI: https://doi.org/10.1002/jcp.70067
HIFNAWY, M. S. et al. Nature-inspired male contraceptive and spermicidal products. Phytochemistry Reviews, v. 20, p. 797–843, 2020.
JOHNSTON, D.; GOLDBERG, E. Preclinical contraceptive development for men and women. Biology of Reproduction, v. 103, n. 2, p. 147–156, 2020. DOI: https://doi.org/10.1093/biolre/ioaa076
KARANWAL, S. et al. Higher abundance of DLD protein in buffalo bull spermatozoa causes elevated ROS production leading to early sperm capacitation and reduction in fertilizing ability. Journal of Animal Science and Biotechnology, v. 15, n. 1, 2024. DOI: https://doi.org/10.1186/s40104-024-01085-6
KARTHIKEYAN, S. et al. Statin drugs and lipid modulation: mechanistic basis considering lipid rafts, kinase signaling, myopathy, and cancer. Pharmacological Research, p. 107912, 2025. DOI: https://doi.org/10.1016/j.phrs.2025.107912
KENT, K. et al. Toward development of the male pill: a decade of potential non-hormonal contraceptive targets. Frontiers in Cell and Developmental Biology, v. 8, p. 61, 2020. DOI: https://doi.org/10.3389/fcell.2020.00061
KHAMAMKAR, A. et al. Disrupting sperm maturation: a non-hormonal contraceptive approach, targeting GSK3α in the epididymal spermatozoa. Human Reproduction, v. 40, supl. 1, 2025. DOI: https://doi.org/10.1093/humrep/deaf097.353
KIM, K. J.; JEON, Y. J.; LEE, S. H. Anti-obesity effects of fucoidan. Journal of Nutritional Biochemistry, v. 25, n. 5, p. 456–463, 2014. DOI: https://doi.org/10.1016/j.jnutbio.2013.12.006
LASTRA-VARGAS, L. M. et al. Endogenous ouabain: a possible link between high salt intake and male infertility. Reproductive System & Sexual Disorders: Current Research, v. 10, p. 1–4, 2021. DOI: https://doi.org/10.35248/2161-038X.21.10.256
LEE, K.; HWANG, J. Ca2+ homeostasis and male fertility: a target for a new male contraceptive system. Animal Cells and Systems, v. 28, n. 4, p. 171–183, 2024. DOI: https://doi.org/10.1080/19768354.2024.2345647
LEE, S. et al. 17BIPHE2, an engineered cathelicidin antimicrobial peptide with low susceptibility to proteases, is an effective spermicide and microbicide against Neisseria gonorrhoeae. Human Reproduction, 2022. DOI: https://doi.org/10.1093/humrep/deac188
LI, B.; LU, F.; WEI, X.; ZHAO, R. Fucoidan: structure and bioactivity. Molecules, v. 13, n. 8, p. 1671–1695, 2008. DOI: https://doi.org/10.3390/molecules13081671
MARIANI, N. et al. Advances in non-hormonal male contraception targeting sperm motility. Human Reproduction Update, v. 29, n. 5, 2023. DOI: https://doi.org/10.1093/humupd/dmad008
MATEO-OTERO, Y. et al. Embryo development is impaired by sperm mitochondrial-derived ROS. Biological Research, v. 57, n. 1, 2024. DOI: https://doi.org/10.1186/s40659-024-00483-4
MCDERMOTT, J. P. et al. Green fluorescence protein driven by the Na,K-ATPase α4 isoform promoter is expressed only in male germ cells of mouse testis. Journal of Assisted Reproduction and Genetics, v. 29, n. 12, p. 1313–1325, 2012. DOI: https://doi.org/10.1007/s10815-012-9876-x
MCDERMOTT, J. P. et al. Na,K-ATPase α4, and not Na,K-ATPase α1, is the main contributor to sperm motility, but its high ouabain binding affinity site is not required for male fertility in mice. Journal of Membrane Biology, v. 254, n. 5, p. 549–561, 2021. DOI: https://doi.org/10.1007/s00232-021-00181-2
MILEWSKI, D.; JAMES, P. Alpha4 Na,K-ATPase localization and expression are dynamic aspects of spermatogenesis and in sperm incubated under capacitating conditions. International Journal of Molecular Sciences, v. 26, n. 5, p. 1817, 2025. DOI: https://doi.org/10.3390/ijms26051817
MO, L. et al. The mechanism of oxidative stress in asthenozoospermia and antioxidant strategies: a review. Frontiers in Endocrinology, v. 16, p. 1670762, 2025. DOI: https://doi.org/10.3389/fendo.2025.1670762
OISHEE, M. et al. Na+,K+-ATPase α isoforms in sperm show a highly structured and distinct pattern of distribution. Reproduction, v. 170, n. 3, 2025. DOI: https://doi.org/10.1530/rep-25-0114
OLIVEIRA, J. et al. Glycerol kinase 2 as a metabolic sentinel for human sperm motility and male fertility. Biomolecules, v. 15, n. 9, p. 1249, 2025. DOI: https://doi.org/10.3390/biom15091249
PATANKAR, M. S. et al. A revised structure for fucoidan may explain some of its biological activities. Journal of Biological Chemistry, v. 268, n. 29, p. 21770–21776, 1993.
PRACTICE COMMITTEE OF THE AMERICAN SOCIETY FOR REPRODUCTIVE MEDICINE. Combined hormonal contraception and the risk of venous thromboembolism: a guideline. Fertility and Sterility, v. 107, n. 1, p. 43–51, 2017. DOI: https://doi.org/10.1016/j.fertnstert.2016.09.027
PRADHAN, B. et al. Multifunctional role of fucoidan, sulfated polysaccharides in human health and disease: a journey under the sea in pursuit of potent therapeutic agents. International Journal of Biological Macromolecules, v. 164, p. 4263–4278, 2020. DOI: https://doi.org/10.1016/j.ijbiomac.2020.09.019
QIU, S. et al. Bioactive polysaccharides from red seaweed as potent food supplements: a systematic review of their extraction, purification, and biological activities. Carbohydrate Polymers, v. 275, p. 118696, 2022. DOI: https://doi.org/10.1016/j.carbpol.2021.118696
RAJASEKARAN, M. et al. Spermicidal activity of an antifungal saponin obtained from the tropical herb Mollugo pentaphylla. Contraception, v. 47, n. 4, p. 401–412, 1993. DOI: https://doi.org/10.1016/0010-7824(93)90037-8
REN, D. et al. Hypolipidemic effects of fucoidan fractions from Saccharina sculpera (Laminariales, Phaeophyceae). International Journal of Biological Macromolecules, v. 140, p. 188–195, 2019. DOI: https://doi.org/10.1016/j.ijbiomac.2019.08.002
RITHAPORN, T.; MONGA, M.; RAJASEKARAN, M. Curcumin: a potential vaginal contraceptive. Contraception, v. 68, n. 3, p. 219–223, 2003. DOI: https://doi.org/10.1016/s0010-7824(03)00163-x
RUZZI, F. et al. Lipid rafts, caveolae, and epidermal growth factor receptor family: friends or foes? Cell Communication and Signaling, v. 22, n. 1, 2024. DOI: https://doi.org/10.1186/s12964-024-01876-4
SABRI, H. et al. The yin and yang of sodium lauryl sulfate use for oral and periodontal health: a literature review. Journal of Dentistry, v. 24, p. 262–276, 2023. DOI: https://doi.org/10.30476/dentjods.2022.95108.1836
SALICIONI, A. et al. Testis-specific serine kinase protein family in male fertility and as targets for non-hormonal male contraception. Biology of Reproduction, v. 103, n. 2, p. 264–274, 2020. DOI: https://doi.org/10.1093/biolre/ioaa064
SANJEEWA, K. et al. Anti-inflammatory mechanisms of fucoidans to treat inflammatory diseases: a review. Marine Drugs, v. 19, 2021. DOI: https://doi.org/10.3390/md19120678
SENGUPTA, P. et al. Oxidative stress affects sperm health and fertility—time to apply facts learned at the bench to help the patient: lessons for busy clinicians. Reproductive Medicine and Biology, v. 23, n. 1, 2024. DOI: https://doi.org/10.1002/rmb2.12598
SONG, X. X. et al. Effect of monosaccharide L-fucose and polysaccharide fucoidan on sperm α-L-fucosidase activity and relation to sperm-oocyte interaction in pig. Asian-Australasian Journal of Animal Sciences, v. 20, n. 3, p. 351–358, 2007. DOI: https://doi.org/10.5713/ajas.2007.351
SUTYAK, K. et al. Spermicidal activity of the safe natural antimicrobial peptide subtilosin. Infectious Diseases in Obstetrics and Gynecology, 2008. DOI: https://doi.org/10.1155/2008/540758
SYEDA, S. et al. Strategies to develop Na,K-ATPase-α4 inhibitors as male contraceptives. International Journal of Molecular Sciences, v. 26, n. 12, p. 5646, 2025. DOI: https://doi.org/10.3390/ijms26125646
SYEDA, S. S. et al. The Na+ and K+ transport system of sperm (ATP1A4) is essential for male fertility and an attractive target for male contraception. Biology of Reproduction, v. 103, n. 2, p. 343–356, 2020. DOI: https://doi.org/10.1093/biolre/ioaa093
TANPHAICHITR, N. et al. Potential use of antimicrobial peptides as vaginal spermicides/microbicides. Pharmaceuticals, v. 9, 2016. DOI: https://doi.org/10.3390/ph9010013
THUNDATHIL, J. C.; RAJAMANICKAM, G. D.; KASTELIC, J. P. Na/K-ATPase and regulation of sperm function. Animal Reproduction, v. 15, supl. 1, p. 711–720, 2018. DOI: https://doi.org/10.21451/1984-3143-AR2018-0024
WANG, T. et al. Core fucosylation regulates the ovarian response via FSH receptor during follicular development. Journal of Advanced Research, v. 67, p. 105–120, 2024. DOI: https://doi.org/10.1016/j.jare.2024.01.025
WANG, Y.; FU, X.; LI, H. Mechanisms of oxidative stress-induced sperm dysfunction. Frontiers in Endocrinology, v. 16, p. 1520835, 2025. DOI: https://doi.org/10.3389/fendo.2025.1520835
WOO, A. L.; JAMES, P. F.; LINGREL, J. B. Sperm motility is dependent on a unique isoform of the Na,K-ATPase. Journal of Biological Chemistry, v. 275, n. 27, p. 20693–20699, 2000. DOI: https://doi.org/10.1074/jbc.M002323200
WORLD HEALTH ORGANIZATION. WHO laboratory manual for the examination and processing of human semen. 6. ed. Geneva: WHO, 2021.
WU, Y. et al. A high-resolution N-glycoproteome landscape of aging mouse ovary. Redox Biology, v. 81, p. 103584, 2025. DOI: https://doi.org/10.1016/j.redox.2025.103584
XU, M. et al. Effects of nonoxynol-9 (N-9) on sperm functions: systematic review and meta-analysis. Reproduction and Fertility, v. 3, n. 1, p. R19–R33, 2022. DOI: https://doi.org/10.1530/RAF-21-0024
XU, Z. et al. Mitochondrial regulation of spermatozoa function: metabolism, oxidative stress and therapeutic insights. Animals, v. 15, n. 15, p. 2246, 2025. DOI: https://doi.org/10.3390/ani15152246
YANG, Y. H. et al. Relationship between ouabain and asthenozoospermia. Journal of Huazhong University of Science and Technology [Medical Sciences], v. 34, n. 1, p. 87–90, 2014. DOI: https://doi.org/10.1007/s11596-014-1236-x
YE, W. et al. Spin and wavelength multiplexed nonlinear metasurface holography. Nature Communications, v. 7, n. 1, p. 11930, 2016. DOI: https://doi.org/10.1038/ncomms11930
YONIS, H. et al. Association of contemporary hormonal contraception and the risk of arterial thrombosis. European Heart Journal, v. 45, supl. 2, 2024. DOI: https://doi.org/10.1093/eurheartj/ehae666.1535
ZAHARIEV, N.; KATSAROV, P.; LUKOVA, P.; PILICHEVA, B. Fucoidan pharmaceutical formulations. Polymers, v. 15, n. 15, p. 3242, 2023. DOI: https://doi.org/10.3390/polym15153242
ZAYED, A. et al. Phenomenological investigation of the cytotoxic activity of fucoidan isolated from Fucus vesiculosus. Process Biochemistry, v. 84, p. 158–165, 2019.
ZHANG, R. et al. Proteomics and metabolomics analyses of mechanism underlying bovine sperm cryoinjury. BMC Genomics, v. 26, n. 1, p. 11258, 2025. DOI: https://doi.org/10.1186/s12864-025-11258-w
ZHERNOV, Y. V. et al. Microbicides for topical HIV immunoprophylaxis: current status and future prospects. Pharmaceuticals (Basel, Switzerland), v. 17, n. 6, p. 668, 2024. DOI: https://doi.org/10.3390/ph17060668
ZHU, Z. et al. Negative effects of ROS generated during linear sperm motility on gene expression and ATP generation in boar sperm mitochondria. Free Radical Biology and Medicine, v. 141, p. 159–171, 2019. DOI: https://doi.org/10.1016/j.freeradbiomed.2019.06.018
License
Copyright (c) 2026 RECIMA21 - Revista Científica Multidisciplinar - ISSN 2675-6218
This work is licensed under a Creative Commons Attribution 4.0 International License.
Os direitos autorais dos artigos/resenhas/TCCs publicados pertecem à revista RECIMA21, e seguem o padrão Creative Commons (CC BY 4.0), permitindo a cópia ou reprodução, desde que cite a fonte e respeite os direitos dos autores e contenham menção aos mesmos nos créditos. Toda e qualquer obra publicada na revista, seu conteúdo é de responsabilidade dos autores, cabendo a RECIMA21 apenas ser o veículo de divulgação, seguindo os padrões nacionais e internacionais de publicação.