The effect of molecular hydrogen and protein kinase C inhibitor on the functional parameters of bovine sperm

Keywords: molecular hydrogen, spermatozoa, staurosporin, proteinkinase C, malondialdehyde, motility, ATP

Abstract

Background.  Studies on the use of molecular hydrogen as a cryoprotector for cattle spermatozoa have shown positive results, an increase in cell mobility and viability, and stabilization of their membranes have been noted. Despite this, the mechanisms of action of molecular hydrogen on spermatozoa remain unclear.

Purpose. Investigation of the mechanisms of action of molecular hydrogen and the inhibitor of protein kinase C – staurosporin on the functional activity of bovine sperm.

Materials and methods. The object of the study was semen samples from Holstinized bulls, subjected to various treatments. Sperm diluted with a standard diluent "BioXcell" was used; sperm diluted with a diluent "BioXcell" enriched with molecular hydrogen; sperm incubated using staurosporin – a protein kinase C inhibitor; sperm treated with molecular hydrogen and subsequent incubation with staurosporin. For each sample in the sperm, an analysis of kinetic parameters, energy status and intensity of free radical processes was carried out.

Results. The inhibitor of proteinkinase C – staurosporin reduced the metabolic and kinetic parameters of spermatozoa, which confirms the direct participation of proteinkinase C in maintaining the structural and functional integrity and activity of spermatozoa. Molecular hydrogen in the presence of staurosporin had a positive effect on the studied parameters of bovine sperm, therefore, in addition to proteinkinase C, other mechanisms that are insensitive to staurosporin are involved in the transduction of molecular hydrogen-induced signals.

Conclusion. Identification of the mechanisms determining the stimulating effects of molecular hydrogen on bull sperm will improve the parameters of fresh sperm and the technology of cryopreservation of bull sperm.

EDN: BAOVMS

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Author Biographies

Marina N. Ivashchenko, Nizhny Novgorod State Agrotechnological University named after L.Y. Florentyev

Cand. Sc. (Biology), Associate Professor

Anna V. Deryugina, National Research Lobachevsky State University of Nizhny Novgorod

Dr. Sc. (Biology), Associate Professor

Andrey A. Belov, Nizhny Novgorod State Agrotechnological University named after L.Y. Florentyev

Cand. Sc. (Biology), Associate Professor

Pavel S. Ignatiev, Production Association "Ural Optical and Mechanical Plant" named after E.S. Yalamov

Cand. Sc. (Phys.-Math.)

Mikhail I. Latushko, Production Association "Ural Optical and Mechanical Plant" named after E.S. Yalamov

Cand. Sc. (Engineering)

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O’Connell, M., McClure, N., Lewis, S.E.M. (2002). The effects of cryopreservation on sperm morphology, motility and mitochondrial function. Hum Reprod, 17, 704-709. https://doi.org/10.1093/humrep/17.3.704

Ohno, K., Ito, M., Ichihara, M., Ito, M. (2012). Molecular hydrogen as an emerging therapeutic medical gas for neurodegenerative and other diseases. Oxid Med Cell Longev, 24, 353152. https://doi.org/10.1155/2012/353152

Ohsawa, I., Ishikawa, M., Takahashi, K., Watanabe, M., Nishimaki, K., Yamagata, K. (2007). Hydrogen acts as a therapeutic antioxidant by selectively reducing cytotoxic oxygen radicals. Nat Med, 13(6), 688-694. https://doi.org/10.1038/nm1577

Paoli, D., Lombardo, F., Lenzi, A. (2014). Sperm cryopreservation: effects on chromatin structure. Adv Exp Med Biol, 791, 137-150. https://doi.org/10.1007/978-1-4614-7783-9_9

Pahune, P.P., Choudhari, A.R., Muley, P.A. (2013). The total antioxidant power of semen and its correlation with the fertility potential of human male subjects. J Clin Diagn Res, 7(6), 991-995. https://doi.org/10.7860/JCDR/2013/4974.3040

Paudel, B., Gervasi, M.G., Porambo, J., Caraballo, D. (2018). Sperm capacitation is associated with phosphorylation of the testis-specific radial spoke protein Rspha. Biol Reprod, 100(2), 440-454. https://doi.org/10.1093/biolre/ioy202

Qiu, P., Liu, Y., Zhang, J. (2019). Recent advances in studies of molecular hydrogen against sepsis. Int J Biol Sci, 15(6), 1261. https://doi.org/10.7150/ijbs.30741

Roten, R., Paz, G. F., Homonnai, Z. T. (1992). Ca2±independent induction of acrosome reaction by protein kinase C in human sperm. Endocrinology, 131(5), 2235-2243. https://doi.org/10.1210/endo.131.5.1425422

Ward, N. E., O’Brian, C. A. (1992). Kinetic analysis of protein kinase C inhibition by staurosporine: evidence that inhibition entails inhibitor binding at a conserved region of the catalytic domain but not competition with substrates. Mol Pharmacol, 41(2), 387-392. https://doi.org/10.1016/S0026-895X(25)08889-3


Published
2025-08-31
How to Cite
Ivashchenko, M., Deryugina, A., Belov, A., Ignatiev, P., & Latushko, M. (2025). The effect of molecular hydrogen and protein kinase C inhibitor on the functional parameters of bovine sperm. Siberian Journal of Life Sciences and Agriculture, 17(3), 43-55. https://doi.org/10.12731/2658-6649-2025-17-3-1225
Section
Human and Animal Physiology