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.
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