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

University of Ulster, Aontroim, Northern Ireland, United Kingdom
Human Reproduction (Impact Factor: 4.57). 06/1998; 13(5):1240-7. DOI: 10.1093/humrep/13.5.1240
Source: PubMed


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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Available from: Valerie McKelvey-Martin, Oct 04, 2015
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    • "In the same way, Long and Kramer (2003) demonstrated that the increase of lipid peroxidation during storage was unaffected by the addition of Vitamin E. However, other reports suggested a positive response of -tocopherol on equine semen during storage for 48 h at 5 • C (Ball and Vo, 2002) and in poultry (Donoghue and Donoghue, 1997). Hughes et al. (1998) found that in vitro treatment of sperm with 3 and 60 M -tocopherol reduced the magnitude of DNA damage. Jeong et al. (2009) suggested that tocopherol supplementation at 200 g/ml may protect sperm against excessive ROS generation by reducing lipid peroxidation and lowering the expression of apoptosis genes by reducing DNA fragmentation. "
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    ABSTRACT: Due to its high antioxidant content, the argan oil could play a beneficial role in liquid storage of ram semen. The aim of this study was to investigate effects of different concentration of argan oil (ARO) on spermatologic parameters, lipid peroxidation and DNA fragmentation during liquid storage of ram semen until 48h. Also effects of extenders and temperature on same parameters were assessed. For these aims, semen samples were collected from Boujaâd rams, extended with Tris egg yolk or skim milk extenders without (control) or supplemented with different concentrations of ARO (1%, 2%, 5% and 10% v/v) at a final concentration of 0.8×10(9) sperm/mL and stored until 48h at 5°C or 15°C. The sperm quality assessments were performed at different intervals during storage (0, 8, 24 and 48h). Sperm progressive motility started to decrease after 8h of storage in all temperatures - extenders combinations and dropped steadily during the 8-48h interval. However, sperm viability, progressive motility and membrane integrity were markedly higher in ARO groups (especially in 1% in Tris and 5% in skim milk) until 24h and 48h storage at both temperatures compared to controls. The argan oil also decreased the level of spontaneous and induced malondialdehyde (MDA) and the sperm DNA fragmentation until 48h storage. In conclusion, it was determined that addition of argan oil to conventional extenders may improve the quality of ram semen during liquid storage in different temperatures. Copyright © 2015 Elsevier B.V. All rights reserved.
    Animal reproduction science 07/2015; 160. DOI:10.1016/j.anireprosci.2015.07.003 · 1.51 Impact Factor
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    • "The involvement of ROS in male fertility is due to its capacity to induce detrimental chemical and structural modifications to sperm nuclear DNA, damage to the proteins and lipids in sperm and mitochondrial-membranes (Gharagozloo and Aitken, 2011). Antioxidants play an essential role in the maintenance of the motility and genetic integrity of spermatozoa (Hughes et al., 1998; Mata-Campuzano et al., 2012). Progressive motility is a sensitive parameter for detecting abnormal sperm motion (Horimoto et al., 2000). "
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    ABSTRACT: Background: Diabetes mellitus characterized by hyperglycaemia could affect sperm quality as a result of increased oxidative stress. This study was performed to investigate the effects of red palm oil (RPO), aqueous rooibos tea extracts (RTE) as well as their combination (RPO + RTE) on sperm motility parameters in streptozotocin-induced diabetic rats. Materials and methods: Diabetes was induced by a single administration of streptozotocin (50 mg/kg) and the rats were treated with red palm oil (2 ml/day) and / or aqueous rooibos tea extract (2%) for 7 weeks. Sperm motility parameters were measured using Computer Assisted Sperm Analyzer (CASA). Results: Hyperglycaemia negatively affected the sperm progressive motility significantly at p<0.05. There was a significant decrease (p<0.05) in sperm linearity (LIN) in the diabetic group when compared with the normal control group. RPO supplemented diabetic rats exhibited increased progressive sperm motility, sperm linearity (LIN) and wobble (WOB). Significant decreases (p<0.05) in straight line velocity (VSL) and average path velocity (VAP) of the sperms were observed in all the diabetic groups when compared to the control group. Significant (p<0.05) elevated levels of WOB and LIN were observed following RTE treatment and co-administration with RPO respectively. Conclusion: The present study suggests that red palm oil and / or rooibos administration exhibited no adverse effects on sperm motility parameters but rather showed some beneficial effects.
    African Journal of Traditional, Complementary and Alternative Medicines 09/2014; 11(5):8-15. DOI:10.4314/ajtcam.v11i5.2 · 0.56 Impact Factor
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    • "It is possible that the free radical scavengers within the semen extender were sufficient to protect the spermatozoa from oxidative stress generated during cryopreservation. In fact, the normal sperm DNA is less susceptible to this process (Zini et al., 2009), and in vitro antioxidant supplementations to normal DNA integrity spermatozoa have a limited consequence in DNA protection from ROS (Hughes et al., 1998; Donnelly et al., 1999; 2000; Chi et al., 2008). "
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    ABSTRACT: Cryopreservation of epididymal sperm is an effective technique to preserve genetic materials of domestic cats and wild felids when they unexpectedly die. However, this technique inevitably causes detrimental changes of cryopreserved-thawed spermatozoa, for example, by physical damage and excessive oxidative stress. L-carnitine is an antioxidant that has been used to improve sperm motility in humans and domestic animals. This study aimed to investigate the effects of L-carnitine on cat epididymal sperm quality following cryopreservation and thawing. After routine castration, cauda epididymides were collected from 60 cat testes. The epididymal spermatozoa from 3 cauda epididymides were pooled as 1 replicate. Spermatozoa samples (16 replicates) were examined for spermatozoa quality and then randomly divided into 4 groups: 0 mM L-carnitine (control), 12.5 mM, 25 mM and 50 mM L-carnitine. The sperm aliquots were then equilibrated and conventionally frozen. After thawing, sperm motility, plasma membrane integrity, DNA integrity and acrosome integrity were evaluated. The 25 mM L-carnitine significantly improved sperm motility compared with a control group (p<0.05), although this was not significantly different among other concentrations. In conclusion, supplementation of 25 mM L-carnitine in freezing extender improves cauda epididymal spermatozoa motility. The effects of L-carnitine on the levels of oxidative stress during freezing and thawing remains to be examined.
    Asian Australasian Journal of Animal Sciences 06/2014; 27(6):791-6. DOI:10.5713/ajas.2013.13565 · 0.54 Impact Factor
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