Supraphysiological ROS levels can lead to apoptosis, lipid peroxidation, and DNA and protein damage. This pilot study aimed to investigate the sperm oxidative damage in subfertile men, to describe the relationship between the antioxidant system and ROS. Sixty‐four semen samples were categorised according to the evaluated routine parameters (WHO, WHO laboratory manual for the examination and processing of human semen, 2010). Results were cross‐referenced with the DNA damage [Comet (n = 53) and TUNEL (n = 49) assays], antioxidant enzyme activity [SOD (n = 51), CAT (n = 48) and GST (n = 48)], and content of total thiols (n = 36), lipid hydroperoxides (n = 35) and MDA (n = 31). Compared to pathospermic samples, normozoospermic presented 40%–45% fewer spermatozoa with fragmented DNA, 19% fewer hydroperoxides, and slightly higher total thiols and MDA levels. Asthenozoospermic/asthenoteratozoospermic samples had the lowest GST activity. SOD and CAT showed a similar trend. Our results evidenced significant positive correlations between DNA damage and immotile spermatozoa; SOD and CAT, GST and total thiols; CAT and GST; total thiols and sperm concentration; and MDA levels and head/midpiece abnormalities and hydroperoxides. This work contributes to the existing body of knowledge by showing that the oxidative status correlates with the classic sperm analysis parameters. Oxidative stress and DNA damage evaluation might be a valuable diagnostic and prognostic tool in cases of idiopathic male subfertility.
This study aimed to evaluate the impact of a vapor freezing protocol on antioxidant enzyme activity (superoxide dismutase (SOD) and glutathione reductase (GR)), sperm with active mitochondria, DNA damage, and spermatic parameters. Twenty-six semen samples from men undergoing infertility investigation were cryopreserved in liquid nitrogen (LN) vapors and plunged into LN, with (method A) and without (method B) a commercial sperm freezing medium (SFM) and inherent removal with a sperm preparation medium (SPM). Most parameters were assessed before and after freezing, except for SOD and GR activity, which were only assessed after freezing. Although method A promoted better results than method B, the percentage of spermatozoa with active mitochondria, motility, vitality, and normal morphology decreased significantly. DNA damage (determined by comet assay) increased similarly with both methods, but the percentage of spermatozoa with fragmented DNA (by TUNEL assay) remained similar to fresh values when method A was applied. GR activity was higher and SOD activity lower with method A. The addition of SFM coupled with the sperm wash with SPM seems essential to preserve the quality of most of the analyzed spermatic parameters and active mitochondria. The detrimental effects promoted by freezing were shown to depend on the quality of the fresh semen, according to correlation coefficients. Interestingly, thawed samples of both methods shared similar DNA damage. These results highlight the need to find more effective protocols, especially for the freezing of low-quality semen samples.
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