The effects of antioxidant supplementation during Percoll preparation on human sperm DNA integrity

ISSN： 1460-2350

Source： Human Reproduction, Vol.13, Iss.5, 1998-05, pp. : 1240-1247

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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 μM), alpha tocopherol (30, 60 μM), urate (200, 400 μM), or acetyl cysteine (5, 10 μM). 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 μM), alpha tocopherol (30 and 60 μM) and urate (400 μM). 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. Keywords: alpha tocopherol/ascorbic acid/comet assay/DNA damage/urate