Time course of hydrogen peroxide induced changes in the lipid peroxidation of human sperm membranes
Bell, M.; Sikka, S.; Rajasekaran, M.; Hellstrom, W.
Advances in Contraceptive Delivery Systems Cds 8(1-2): 144-150
1992
ISSN/ISBN: 1012-8689 PMID: 12285560 Document Number: 537388
The present study evaluates the changes in sperm motility due to H-2O-2 induced membrane damage. Washed human sperm suspended in HAM's F-10 (20-30 x 10-6/ml) were incubated (37 degree C) with varying concentrations (0-0.05%) of H-20-2 for up to 15 minutes. Sperm were analyzed for percent motility, percent 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 choroform:methanol (1:2) and used in colorimetric determination of cholesterol and phospholipid contents (mu-mol/10-6 sperm). Lipid peroxidation was measured by the production of malondialdehyde (nmol MDA/10-8 sperm). The results (mean +- SEM, n = 8) indicate a dose and time-dependent effect on % motility during the 15 minute incubation period. In comparison to control (8 +- 4%), samples incubated with 0.01% H-20-2 exhibited a 25 +- 3% decrease in % motility, while a complete loss of motility was observed with 0.05% H-20-2. No significant differences in decrease in sperm viability were observed between control (11 +- 4) and H-20-2 (0.01%) treated samples (14 +- 2%). An increase of (54 +- 5%) in lipid peroxidation was observed with 0.01% H-20-2, as compared with a 18+-1% increase in control samples at 15 minutes. The C/PL ratio increased by 46+-4% at 15 minutes in H-20-2 treated samples while showing a 34.3% decrease in control samples. H-20-2 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.