Hydrogen Peroxide Has Adverse Effects on Human Sperm Quality Parameters, Induces Apoptosis, and Reduces Survival
Dwi Ari Pujianto, Mona Oktarina, Ida Ayu Sharma Sharaswati, Yulhasri
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.