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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 (H2O2) 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 H2O2 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 H2O2 concentrations of 200 and 250 μM. H2O2 reduced sperm capability to penetrate cervical mucus and also damage cell membrane integrity at all concentrations used. H2O2 significantly inhibited sperm capacitation, indicated by reduced total tyrosine phosphorylation. H2O2 exposure stimulated activation of caspase 3 and significantly reduced phosphorylated AKT at all concentrations used. Conclusions: H2O2 comprehensively inhibits sperm qualities related to the capacity to fertilize oocyte, stimulates caspase activity, and inhibits cell survival. [ABSTRACT FROM AUTHOR]

Objective: Progesterone receptor is present in human sperm. Here, we determined if progesterone (P4) improves capacitation and stimulates sperm survival by suppressing apoptosis to evaluate its relevance to be used as a preservation agent to increase and prolong sperm viability and motility. Materials and Methods: Semen samples were obtained from healthy human donors. The sample was washed by Percoll gradient centrifugation and was divided into five groups of 500 µL, each containing approximately 10 million sperm cells. P4 was added to each group at the final concentrations of 0 (control), 250, 500, 750, and 1000 ng/mL, respectively, and was incubated at 37°C for 2 h. Sperm motility was assessed using a computer-assisted sperm analyzer. Tyrosine phosphorylation, Akt phosphorylation, and caspase-3 activation in sperm were analyzed by western immunoblotting. Results: P4 did not increase the sperm motility. Western blot analysis revealed that P4 at the concentration of 500 ng/mL increased tyrosine phosphorylation, indicating increased sperm capacitation. P4 at 750 mg/ml also increased Akt phosphorylation. Interestingly, the activation of Akt was associated with a decrease in caspase-3 activity, indicating the prosurvival effect of P4. Conclusion: This study results suggest that P4 does not influence sperm motility; however, it increases sperm tyrosine phosphorylation (at a specific concentration of 500 ng/ul), activates AKT (Protein kinase B), and suppresses caspase-3 (at a specific concentration of 750 ng/ul). The biochemical effects of P4 on sperm may warrant its use as a preservation agent to increase sperm viability. [ABSTRACT FROM AUTHOR]