Human sperm superoxide anion generation and correlation with semen quality in patients with male infertility

Objective: To measure levels of reactive oxygen species (ROS) including H2O2 and O-2(.-) generation in infertile men and determine whether sperm quality is correlated with levels of ROS triggered by the exogenous reduced form of beta nicotinamide adenine dinucleotide phosphate (NADPH). Design: Prospective study. Setting: Male infertility clinic at a tertiary healthcare center. Patient(s): Eleven infertile men and six healthy donors. Intervention(s): Chemiluminescence assay using luminol and lucigenin as probes before and after incubating sperm samples with 5 mM and 10 mM of NADPH. Main Outcome Measure(s): The ROS generation (10(6) counted photons per minute/10(6) sperm). Result(s): Baseline levels of O-2(.-) generation were significantly higher in the infertile patients than in the healthy donors (r = 0.73, 95% confidence interval [median (25th, 75th percentiles): 0.73 (0.5, 5.5) vs. 0.2 (0.0, 0.5))] when lucigenin was used as the probe. Compared with basal levels, O-2(.-) generation was significantly higher after coincubation with NADPH (5 mM and 10 mM) in the entire combined study population, and patients only but not donors. The O-2(.-) generation was negatively correlated with sperm concentration (r = -0.75, 95% CI 0.38-1), motility (r = -0.69, 95% CI 0.28-1), and percentage of normal morphology (r = -0.78, 95% CI 0.36-1). Conclusion(s): Spermatozoa from infertile men produce higher levels of O-2(.-) in the presence of exogenous NADPH compared to healthy donors. The ability of spermatozoa to generate O-2(.-) increases as the semen quality declines. (C) 2004 by American Society for Reproductive Medicine.