Glutathione s-transferase as a marker of oxidative stress in human ejaculated spermatozoa from patients with pathospermia
AbstractIt is believed that the most common causes of male infertility are impairment of spermatogenesis and sperm functions. Glutathione S-transferases (EC 188.8.131.52) play an important role in sperm physiology, specifically in antioxidant protection against oxidative damage. The catalase decomposition of lipid hydro-peroxides forms as a result of oxidative stress. We used a model of superoxide anion-generating system Fe3+/ascorbate or H2O2-induced stress to study the activity of glutathione s transferase in human ejaculated spermatozoa from patients with pathospermia and products of lipid peroxidation (TBARS) as a marker of oxidative stress. In the present study, dose dependent increase in the level of lipid peroxidation was observed for treatment with Fe3+/ascorbate or H2O2. The TBARS level was higher for sperm cells incubated with superoxide anion-generating system Fe3+/ascorbate than for H2O2. GSTs activity increased in spermatozoa treated with increasing concentration of superoxide anion-generating system Fe3+/ascorbate and H2O. We found that both Fe3+/ascorbate and H2O2 displayed similar inhibitory effects on sperm GSTs activity, however H2O2 at low concentrations activated enzyme activity only in normozoospermic samples, which can be explained as a defence response to oxidative stress. The time course of incubation with 100 μM H2O2 showed a sharp decrease in the enzyme activity during the first 5 min of incubation for both normozoospermic and pathozoospermic men. Preincubation of spermatozoa with GSH completely prevented the ROS-induced inhibition on GSTs only in normozoospermic samples. On the other hand, in pathospermic samples protectory effect of GSH was observed only against non-radical (H2O) radical, but not against radical (superoxide anion-generating system Fe3+/ascorbate) species. The results of our study showed higher oxygen-free radical production, evidenced by increased TBARS level in spermatozoa obtained from infertile men than normozoospermic men. The inhibitory effect of the radical (superoxide anion-generating system Fe3+/ascorbate) species on sperm GSTs activity and products of lipid peroxidation in sperm cells of fertile and infertile men were more expressed compared to non-radical (H2O) species. Our results indicate that estimation of sperm GSTs enzyme assays can be used as a bioindicator for impaired male fertility. The obtained results argue for a biological role of sperm GSTs in susceptibility of spermatozoa to oxidative damage and maintaining sperm antioxidant status.
Aitken, R. J., Jones, K. T., & Robertson, S. A. (2012). Reactive oxygen species and sperm function – in sickness and in health. Journal of Andrology, 33(6), 1096–1106.
Aydemir, B., Onaran, I., Kiziler, A. R., Alici, B., & Akyolcu, M. C. (2007). Increased oxidative damage of sperm and seminal plasma in men with idiopathic infertility is higher in patients with glutathione S-transferase Mu-1 null genotype. Asian Journal of Andrology, 9(1), 108–115.
Ball, B. A., Vo, A. T., & Baumber, J. (2001). Generation of reactive oxygen species by equine spermatozoa. American Journal of Veterinary Research, 62(4), 508–515.
Buzadzic, B., Vucetic, M., Jankovic, A., Stancic, A., Korac, A., Korac, B., & Otasevic, V. (2015). New insights into male (in)fertility: The importance of NO. Brazilian Journal of Pharmacology, 172(6), 1455–1467.
Chen, S. S., Chang, L. S., Chen, H. W., & Wei, Y. H. (2002). Polymorphisms of glutathione S-transferase M1 and male infertility in Taiwanese patients with varicocele. Human Reproduction, 17, 718–725.
Cheng, H., Tchaikovskaya, T., Tu, Y., Chapman, J., Qian, B., Ching, W., Tien, M., Rowe, J. D., Patskovsky, Y. V., Listowsky, I., & Tu, C. D. (2001). Rat glutathione S-transferase M4-4: An isoenzymes with unique structural features including a redox-reactive cysteine-115 residue that forms mixed disulphides with glutathione. Biochemical Journal, 356, 403–441.
Esteves, S. C., & Agarwal, A. (2011). Novel concepts in male infertility. International Brazilian Journal of Urology, 37, 5–715.
Esteves, S. C., & Chan, P. (2015). A systematic review of recent clinical practice guidelines and best practice statements for the evaluation of the infertile male. International Urology and Nephrology, 47, 1441–1456.
Fafula, R. V., Meskalo, О. I., Lychkovskyy, E. I., & Vorobets, Z. (2017). Efekty gidrogen peroksydu na osnovni kinetychni parametry gidrolizu ATR ubai'nchutlyvoju Ca2+,Mg2+-ATR-azoju spermatozoi'div infertyl'nyh cholovikiv [Effect of hydrogen peroxide on the main kinetic parameters of ATP hydrolysis by Ca2+,Mg2+-ATP-ase in spermatozoa of infertile men]. Experimental and Clinical Physiology and Biochemistry, 80(4), 46–51 (in Ukrainian).
Fafula, R. V., Оnufrovych, О. K., & Iefremova, U. P. (2017). Intensyvnist' procesiv lipoperoksydacii' u spermatozoi'dah cholovikiv iz porushennjam [Intensity of processes of lipid peroxidation in sperm of men with impaired fertility ]. Bulletin of Problems Biology and Medicine, 135(1), 199–204 (in Ukrainian).
Farombi, E. O., Adelowo, O. A., & Ajimoko, Y. R. (2007). Biomarkers of oxidative stress and heavy metal levels as indicators of environmental pollution in African cat fish (Clarias gariepinus) from Nigeria Ogun River. International Journal of Environmental Research and Public Health, 4(2), 158–165.
Garrido, N., Meseguer, M., Simon, C., Pellicer, A., & Remohi, J. (2004). Prooxidative and antioxidative imbalance in human semen and its relation with male fertility. Asian Journal of Andrology, 6, 59–65.
Halliwell, B. (2007). Biochemistry of oxidative stress. Biochemical Society Transactions, 35(5), 1147–1150.
Halliwell, B., & Gutteridge, J. (2007). Free radicals in biology and medicine. Oxford University Press, Oxford.
Hayes, J. D., Flanagan, J. U., & Jowsey, I. R. (2005) Glutathione transferases. Annual review of pharmacology and toxicology, 45, 51–88.
Hemachand, T., Gopalakrishnan, B., Salunke, D. M., Totey, S. M., & Shaha, C. (2002). Sperm plasma-membrane-associated glutathione S-transferases as gamete recognition molecules. Journal of Cell Science, 115(10), 2053–2065.
Hesham, N., Moemen, L. A., & Abu Elela, M. H. (2008). Studing the levels of malondialdehyde and antioxidant parameters in normal and abnormal human seminal plasma. Australian Journal Basic and Applied Sciences, 2(3), 773–778.
Joffe, M. (2010). What has happened to human fertility? Human Reproduction, 25, 295–307.
Letelier, M. E., Molina-Berríos, A., Cortés-Troncoso, J., Jara-Sandoval, J. A., Müller, A., & Aracena-Parks, P. (2010). Comparative effects of superoxide anion and hydrogen peroxide on microsomal and cytosolic glutathione S-transferase activities of rat liver. Biological Trace Element Research, 134(2), 203–211.
Monaghan, P., Metcalfe, N. B., & Torres, R. (2009). Oxidative stress as a mediator of life history trade-offs: Mechanisms, measurements and interpretation. Ecology Letters, 12, 75–92.
Nissar, S., Sameer, A. S., Rasool, R., Chowdri, N. A., & Rashid, F. (2017). Glutathione S transferases: Biochemistry, polymorphism and role in colorectal carcinogenesis. Journal of Carcinogenesis and Mutagenesis, 8, 2.
Olshan, A. F., Luben, T. J., & Hanley, N. M. (2010). Preliminary examination of polymorphisms of GSTM1, GSTT1, and GSTZ1 in relation to semen quality. Mutation Research – Fundamental and Molecular Mechanisms of Mutagenesis, 688(1–2), 41–46.
Onaran, I., Guven, G., Ozaydin, A., & Ulutin, T. (2001). The influence of GSTM1 null genotype on susceptibility to in vitro oxidative stress. Toxicology, 157(3), 195–205.
Sahoo, D. K. (2011). Effects of thyroid hormone on testicular functions and antioxidant defence status. Biochemistry: An Indian Journal, 5(6), 337–334.
Said, T. M., Agarwal, A., Sharma, R. K., Thomas, A. J., & Sikka, S. C. (2005). Impact of sperm morphology on DNA damage caused by oxidative stress induced by beta-nicotinamide adenine dinucleotide phosphate. Fertility and Sterility, 83, 95–103.
Sakkas, D., Seli, E., Bizzaro, D., Tarozzi, N., & Manicardi, G. C. (2003). Abnormal spermatozoa in the ejaculate: Abortive apoptosis and faulty nuclear remodelling during spermatogenesis. Reproductive Biomedicine Online, 7, 428–432.
Storey, B. T. (2008). Mammalian sperm metabolism: Oxygen and sugar, friend and foe. International Journal of Developmental Biology, 52(5–6), 427–437.
Tang, K., Xue, W., Xing, Y., Xu, S., Wu, Q., Liu, R., Wang, X., & Xing, J. (2012). Genetic polymorphisms of glutathione S-transferase M1, T1, and P1, and the assessment of oxidative damage in infertile men with varicoceles from northwestern China. Journal Andrology, 33(2), 257–263.
Timirbulatov, R., & Seleznev, Е. (1981). Metody povyshenija intensivnosti svobodno-zadikal'nogo okislenija lipidosoderzhashhih komponentov krovi i ego diagnosticheskoe znachenie [Method of increasing intensity of free-radical oxidization of lipid components of blood and its diagnostic value]. Laboratory Business, 4, 209–211 (in Russian).
Vorobets, D., & Kocheshkova, N. (2008). Neplіdnіst' ta erektil'na disfunkcіja cholovіkіv: Bіohіmіchnі ta klіnіchnі aspekty [Infertility and erectile dysfunction men: Biochemical and clinical aspects]. Ukrmedknyha, Ternopil (in Ukrainian).
Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons «Attribution» 4.0 License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.