The effects of antioxidant supplementation during Percoll preparation on human sperm DNA integrity |
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Authors: | Hughes CM; Lewis SE; McKelvey-Martin VJ; Thompson W |
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Institution: | Department of Obstetrics and Gynaecology, The Queens University of Belfast, Institute of Clinical Science, UK. |
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Abstract: | The integrity of sperm DNA is crucial for the maintenance of genetic
health. A major source of damage is reactive oxygen species (ROS)
generation; therefore, antioxidants may afford protection to sperm DNA. The
objectives of the study were, first, to measure the effects of antioxidant
supplementation in vitro on endogenous DNA damage in spermatozoa using the
single cell gel electrophoresis (comet) assay and, second, to assess the
effect of antioxidant supplementation given prior to X-ray irradiation on
induced DNA damage. Spermatozoa from 150 patients were prepared by Percoll
centrifugation in the presence of ascorbic acid (300, 600 microM), alpha
tocopherol (30, 60 microM), urate (200, 400 microM), or acetyl cysteine (5,
10 microM). DNA damage was induced by 30 Gy X-irradiation. DNA strand
breakage was measured using the comet assay. Sperm DNA was protected from
DNA damage by ascorbic acid (600 microM), alpha tocopherol (30 and 60
microM) and urate (400 microM). These antioxidants provided protection from
subsequent DNA damage by X-ray irradiation. In contrast, acetyl cysteine or
ascorbate and alpha tocopherol together induced further DNA damage.
Supplementation in vitro with the antioxidants ascorbate, urate and alpha
tocopherol separately has beneficial effects for sperm DNA integrity.
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