Time course of hydrogen peroxide induced changes in the lipid peroxidation of human sperm membranes

The present study evaluates the changes in sperm motility due to H202 induced membrane damage. Washed human sperm suspended in HAM's F-10 (20-30 x 10 6/ml) were incubated (37 degrees Celsius) with varying concentrations (0-0.05%) of H202 for up to 15 minutes. Sperm were analyzed for % motility, % viability, the ratio of cholesterol to phospholipids (C/PL), and the degree of lipid peroxidation (LPO). Motility was monitored manually and viability was evaluated by the Eosin Y staining method. Total lipids were extracted with chloroform:methanol (1:2) and used in colorimetric determination of cholesterol and phospholipid contents (mcmol/106 sperm). Lipid peroxidation was measured by the production of malondialdehyde (nmol MDA/108 sperm). The results (mean +or- SEM, n=8) indicate a dose a time-dependent effect on % motility during the 15 minute incubation period. In comparison to control (8 +or- 4%), samples incubated with 0.01% H202 exhibited a 25 +or- 3% decrease in % motility, while a complete loss of motility was observed with 0.05% H202. No significant differences in decrease in sperm viability were observed between control (211 +or- 4) and H202 (0.01%) treated samples (14 +or- 2%). An increase of (54 +or- 5%) in lipid peroxidation was observed with 0.01% H202, as compared with an 18 +or- 1% increase in control samples at 15 minutes. The C/PL ratio increased by 46 +or- 4% at 15 minutes in H202 treated samples while showing a 34.3% decrease in control samples. H202 inhibited sperm motility while increasing membrane LPO and C/PL, without altering sperm viability. It would appear that lipid peroxidation and alteration of sperm membrane composition lead to the loss of sperm motility.