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

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.