Сharacteristics of Glutathione Peroxidase of Spermatozoa of Infertile Men

Authors

DOI:

https://doi.org/10.29038/2617-4723-2018-381-77-81

Keywords:

glutathione peroxidase, antioxidant system, male infertility, pathospermia

Abstract

It is well known that the unbalancing between the intensity of lipoperoxidation processes and the activity of antioxidant defense systems leads to the development of oxidative stress, which is a potential prerequisite for the development of sperm dysfunction. A special role in the system of antioxidant protection belongs to the glutathione system, in particular glutathione peroxidise, which inactivates the active forms of Oxygen with the simultaneous oxidation of reduced glutathione. The aim of present work is to study the kinetic properties of glutathione peroxidise of spermatozoa in fertile and infertile men with different forms of pathospermia.

The activity of glutathione peroxidise was determined by the amount of reduced glutathione (GSH) that was used to neutralize hydrogen peroxide in the glutathione peroxidase reaction. The apparent kinetic parameters characterizing the glutathione peroxidise reaction - the apparent affinity constant for GSH and the maximum reaction rate were determined in the Lineweaver–Burk plot (or double reciprocal plot).
It was found that increasing the concentration of GSH in the incubation medium leads to a gradual increase in enzymatic activity of glutathione peroxidise reaching the plateau. The maximum activity of glutathione peroxidise of sperm cells in fertile and infertility men was noted in the presence of 5 mM GSH in the incubation medium. The value of maximum reaction rate for glutathione peroxidise of infertility men with different forms of pathospermia was 2,4–4,1 fold lower compared to this value in fertile men with normosospermia. At the same time, there was no statistically significant difference in maximum reaction rate between different forms of pathospermia. The value of the apparent affinity constant for GSH in the spermatozoa of the infertility men was 1,9–4,9 fold higher that of the spermatozoa of the normozoospermic males, indicating a decrease in the enzyme affinity for GSH in the pathospermia.
In interpreting the obtained kinetic parameters determined by GSH, it has been shown that in the pathospermia the inhibition of glutathione peroxidise occurs in a mixed type-both by reducing the reaction rate of the enzyme and by reducing the affinity constant to the substrate.

References

1. Abdallah, B. F.; Fetoui, H.; Zribi, N.; Fakfakh, F.; AmmarKeskes, L. (2011) Antioxidant supplementations in vitro improve rat sperm parameters and enhance antioxidant enzyme activities against dimethoate-induced sperm damages. Andrologia, 44(1), pp 272–279. doi: 10.1111/j.1439-0272.2011.01177
2. de Lamirande, E.; O'Flaherty, C. (2008) Sperm activation: role of reactive oxygen species and kinases. Biochimica et Biophysica Acta, 1784(1), pp 106–115. DOI:10.1016/j.bbapap.2007.08.024
3. Agarwal, A.; Virk, G.; Ong, Ch.; S du Plessis S. (2014) Effect of Oxidative Stress on Male Reproduction. The World Journal of Men's Health, 32(1), pp 1–17. doi: 10.5534/wjmh.2014.32.1.1
4. Aitken, R. J.; Baker, M. A. (2013) Oxidative stress, spermatozoa and leukocytic infiltration: Relationships forged by the opposing forces of microbial invasion and the search for perfection. Journal of Reproductive Immunology, 100(1), pp 11–19. doi: 10.1016/j.jri.2013.06.005
5. Cacciatore, I.; Cornacchia, C; Pinnen, F.; Mollica, A.; Di Stefano, A. (2010) Prodrug approach for
increasing cellular Glutathione levels. Molecules, 15, ( 3), pp 1242–1264. doi: 10.3390/molecules15031242
6. Lavryshyn, Yu. Yu.; Varkholiak, I. S.; Martyshuk, T. V.; Huta, Z. A.; Ivankiv, L. B.; Paladiichuk, O. R.; Murska, S. D.; Hutyi, B. V.; Hufrii, D. F. (2016) Biolohichne znachennia systemy antyoksydantnoho zakhystu orhanizmu tvaryn [Biological value of the system of antioxidant protection of an organism of animals]. Naukovyi visnyk Lvivskoho natsionalnoho universytetu veterynarnoi medytsyny ta biotekhnolohii imeni S. Z. Gzhytskoho, 18, 2(66), s. 100–111. doi:10.15421/nvlvet6622 (in Ukrainian)
7. Morielli, T.; O'Flaherty, C. (2015) Oxidative stress impairs function and increases redox protein modifications in human spermatozoa. Reproduction, 149(1), pp 113–123. doi: 10.1530/REP-14-0240
8. Onufrovych, O. K.; Fafula, R. V.; Vorobets, D. Z. (2013) Stan hlutationovoi antyoksydantnoi systemy spermatozoidiv pry ekskretorno-toksychnii formi neplidnosti cholovikiv [State of glutathione antioxidant system of spermatozoa with excretory-toxic form of infertility of men]. Zdobutky klinichnoi i eksperymentalnoi medytsyny, 2, s. 148-151. (in Ukrainian)
9. Onufrovych, O. K.; Fafula, R. V.; Nakonechnyi, Y. A.; Vorobets, D. Z.; Yefremova, U. P.; Vorobets, Z. D. (2016) Aktyvnist hlutationzalezhnykh enzymiv spermatozoidiv za umov patospermii [The activity of glutathion-dependent enzymes of sperm in conditions of pathospermia]. Medychna ta klinichna khimiia; 18(4), s. 5–10. doi:10.11603/mcch.2410-681X.2016. v0.i4. 7246 (in Ukrainian)
10. WHO Laboratory Manual for the examination and processing of human semen, in 5th ed. World Health Organization Geneva; 2010, 271 p.
11. Moin, V. M. (1986) Prostoj i specificheskij metod opredeleniya aktivnosti glutationperoksidazy v ehritrocitah [A simple and specific method for determining the activity of glutathione peroxidase in erythrocytes]. Laboratornoe delo, 12, s. 124–126. (in Russian)
12. Keleti, T. (1986) Basic enzyme kinetics Academiai Kiado; Budapest, pp 112−129.
13. Muller, F. L.; Lustgarten, M. S.; Jang, Y.; Richardson, A.;Van Remmen, H. (2007) Trends in oxidative aging theories. Free Radical Biology and Medicine, 43(4), pp 477–503 doi:10.1016/j. freeradbiomed.2007.03.034
14. Freedman, J. E.; Frei, B; Welch, G. N.; Loscalzo, J. (1995) Glutathione peroxidase potentiates the inhibition of platelet function by S-nitrosothiols. Journal of Clinical Investigation; 96(1), pp 394–400. doi: 10.1172/JCI118047
15. Concetta, E. A.; Mantilacci, L.; Mauro, N.; Principato, G. (2000) Association of glutathione peroxidase activity with an acidic glutathione S‐transferase in carp liver. Italian Journal of Zoology; 67(1), pp 39–43. doi: 10.1080/11250000009356292

Published

2018-12-26

How to Cite

Fafula, R., Оnufrovych О., Iefremova, U., & Vorobets, Z. (2018). Сharacteristics of Glutathione Peroxidase of Spermatozoa of Infertile Men. Notes in Current Biology, (8(381), 77–81. https://doi.org/10.29038/2617-4723-2018-381-77-81