Hydrogen Peroxide Has Adverse Effects on Human Sperm Quality Parameters, Induces Apoptosis, and Reduces Survival

Abstract

Background: One of the causes of male fertility disorders is the exposure of oxidative stress on the human sperm. Understanding the mechanism of disturbance is important to develop a better treatment for infertile or subfertile patients. Aims: The aim of this study was to analyze the effects of hydrogen peroxide (H 2 O 2 ) on human sperm quality parameters and cell survival. Settings and Design: This study used an experimental design. Materials and Methods: Sperm cells from 15 donors were washed in a Percoll gradient and dissolved in Biggers, Whitter, and Whittingham medium. Cells were incubated with H 2 O 2 at various concentrations from 0 to 250 μM for 2 h. Sperm viability was examined by eosin assay, sperm kinetic by computer-assisted sperm analyzer, sperm penetration by cervical mucus penetration assay, and membrane integrity by hypo-osmotic swelling test. Sperm capacitation, apoptosis, and cell survival were analyzed using western immunoblotting. Statistical Analysis Used: One-way ANOVA on SPSS 21 combined with post hoc LSD test was used to analyze differences among the groups. A P < 0.05 was considered significant. Results: Sperm viability and kinetic were significantly reduced at H 2 O 2 concentrations of 200 and 250 μM. H 2 O 2 reduced sperm capability to penetrate cervical mucus and also damage cell membrane integrity at all concentrations used. H 2 O 2 significantly inhibited sperm capacitation, indicated by reduced total tyrosine phosphorylation. H 2 O 2 exposure stimulated activation of caspase 3 and significantly reduced phosphorylated AKT at all concentrations used. Conclusions: H 2 O 2 comprehensively inhibits sperm qualities related to the capacity to fertilize oocyte, stimulates caspase activity, and inhibits cell survival.

The Impact of Origanum vulgare Supplementation on Human Asthenozoospermic Sperm Parameter Quality

“…Tis positive correlation between CAT activity and sperm motility can be explained by the importance of this enzyme in the protection of sperm from potentially toxic ROS levels [25]. Treatments with H 2 O 2 produced an increase in MDA levels and Te Scientifc World Journal 3 inhibited advanced sperm motility parameters [26]. However, the incubation of human sperm H 2 O 2 revealed that human catalase was able to counteract the adverse efects of H 2 O 2 on sperm motility [12].…”

Oxido-Reduction Potential as a Method to Determine Oxidative Stress in Semen Samples

“…Numerous studies can also be found in the case of H 2 O 2 treatment. In each of these, a decrease in the viability of sperm cells, deterioration of the kinetic parameters, reduced capacitation and penetration ability, and increased lipid peroxidation can be observed [85,86].…”

“…•− formation, H 2 O 2 exposures were applied as a direct OS source, and t-BuOOH treatments were performed to cause the peroxidation of lipids [77,86,88]. The samples of both groups were aliquoted into Eppendorf tubes (300 µL/tube) and the following treatments were performed: (i) 0 mM, 1 mM, 2 mM, and 5 mM H 2 O 2 , (ii) 0 mM, 1 mM, 2 mM, and 5 mM menadione, (iii) 0 mM, 1 mM, 2 mM, and 5 mM t-BuOOH.…”

Arsenic, Oxidative Stress and Reproductive System

“…The overproduction of NO has been observed in infertile women and men with varicocele. Common forms of ROS that affect sperm survival and its function include H 2 O 2 , OH• [ 23 , 24 ], peroxyl radicals, and hypochlorite ions [ 25 , 26 ]. ROS-induced damage to carbohydrates and lipids leads to the formation of ketoamines and ketoaldehydes [ 27 ], alkyl radicals, such as malondialdehyde (MDA), and 4-hydroxynonenal (HNE) (i.e., biomarkers of lipid peroxidation), respectively [ 28 ].…”