Document Type : Original Article
BIOLOGY DEPARTMENT, SHAHREKORD UNIVERSITY SHAHREKORD, IRAN
Introduction: In the present investigation, we aimed to study the effect of different concentrations of ascorbate/vitamin C, on membrane integrity, viability, and acrosome reaction of human spermatozoa in presence of ferrous (Fe2+) ions as lipid peroxidation (LPO) promoter.
Material and Methods: We used 12 semen samples from healthy donors in Shahrekord. The samples were then washed twice with Tyrode's ringer and centrifuged (500 xg, for 10 min) to separate semen plasma from sperm cells (dissolved in Tyrode's ringer). LPO test was performed to assess generated MDA in media spectrophotometrically. Eosin staining method was used to detect dead or alive sperm cells. To assess the ability of sperm cells undergoing acrosome reaction, the gelatin digestion test was performed in which iron and ascorbate were added to sperm cells and stained with comassie blue dye. Number of sperm cells with/without halo around their heads was recorded. We used Hest as statistical analysis.
Results: Ascorbate in concentrations below 1000 micromolar protects spermatozoa from free radical damages as evidenced by improvement in their membrane integrity (data of LPO test), elevated acrosome reactions, and increased viability. Concomitantly, there is also witnessed depletion of malondialdehyde (MDA) (an end product of LPO) generation following ascorbate supplementation. Ascorbate at 1000 micromolar concentration and above, however, is not protective, as evidenced by abrupt fall in sperm membrane integrity and rate of acrosome reactions and lowered percent of alive sperm cells.
Conclusion: Collectively, ascorbate acts as a double-edged sward in different concentrations and researchers must keep in mind this negative side of ascorbate application in their research fields like in vitro fertilization, cell culture, and preservation of gametes to minimize its deleterious impacts on biological systems.