A Comparison Between Two Assays for Measuring Seminal Oxidative Stress and their Relationship with Sperm DNA Fragmentation and Semen Parameters

Abstract

Oxidative stress (OS) is a significant cause of DNA fragmentation and is associated with poor embryo development and recurrent miscarriage. The aim of this study was to compare two different methods for assessing seminal OS and their ability to predict sperm DNA fragmentation and abnormal semen parameters. Semen samples were collected from 520 men attending for routine diagnostic testing following informed consent. Oxidative stress was assessed using either a chemiluminescence assay to measure reactive oxygen species (ROS) or an electrochemical assay to measure oxidation reduction potential (sORP). Sperm DNA fragmentation (DFI) and sperm with immature chromatin (HDS) were assessed using sperm chromatin structure assay (SCSA). Semen analysis was performed according to WHO 2010 guidelines. Reactive oxygen species sORP and DFI are negatively correlated with sperm motility (p = 0.0012, 0.0002, <0.0001 respectively) and vitality (p < 0.0001, 0.019, <0.0001 respectively). The correlation was stronger for sORP than ROS. Reactive oxygen species (p < 0.0001), sORP (p < 0.0001), DFI (p < 0.0089) and HDS (p < 0.0001) were significantly elevated in samples with abnormal semen parameters, compared to those with normal parameters. Samples with polymorphonuclear leukocytes (PMN) have excessive ROS levels compared to those without (p < 0.0001), but sORP and DFI in this group are not significantly increased. DNA fragmentation was significantly elevated in samples with OS measured by ROS (p = 0.0052) or sORP (p = 0.004). The results demonstrate the multi-dimensional nature of oxidative stress and that neither assay can be used alone in the diagnosis of OS, especially in cases of leukocytospermia.

Protective effect of Berberine on reproductive function and spermatogenesis in diabetic rats via inhibition of ROS/JAK2/NFκB pathway

“…32,55 These diseases have rather different mechanisms, whereas their pathogenesis is closely related to oxidative stress. 7,8,56,57 At the same time, the ratio of p-JAK2 and JAK2…”

“…4 Studies have revealed increased oxidative stress in the pathology of DM 5,6 and that an excess amount of reactive oxygen species (ROS) exhibits pathological effects on spermatozoa, resulting in male infertility. 7,8 Janus kinase 2 (JAK2) is an important signaling molecule that is expressed in the testicles. As reported in a previous study, NOX5 and p22phox-derived ROS are involved in the activation of JAK2, which is upstream of the NFκB pathway.…”

Predictive value of oxidative stress testing in semen for sperm DNA fragmentation assessed by sperm chromatin dispersion test

“…24 Currently, the investigation of oxidative stress is based on the detection of intracellular ROS through chemiluminescence assay, 25 total antioxidant capacity (TAC), 26 quantification of the end products of lipid peroxidation (eg, malondialdehyde) 27 or DNA damage (8OHdG) 28 with strong correlations being reported between these different OS markers, and increased SDF in semen samples of infertile men. 19,29,30 Furthermore, SDF negatively affects sperm fertility potential and embryo development in natural conception and assisted reproductive techniques (ART). [31][32][33] In addition, evidence suggests that the assessment of oxidative stress could be important for infertile patients, who could take advantage of antioxidants supplementation or modification of the lifestyle (eg, smoking, exposure to pollutants, varicocoele, high BMI).…”

“…Bars show the standard error of the mean. Patients were subdivided into groups, namely low (≤1.34 mV/10 6 spermatozoa/mL) or high (>1.34 mV/10 6 spermatozoa/mL) ORP, and low (≤30%) or high (>30%) SDF between SDF and seminal ROS levels, but no correlation was observed between SDF and ORP in a small group of 47 patients with unknown fertility 30. In the latter study, the small sample size may be the explanation for these apparently contrasting results.…”

Form from Function, Order from Chaos in Male Germline Chromatin

“…Finally, we are particularly proud of the use of novel methods for studying the interplay between chromatin structure and the susceptibility to DNA damage and mutation. Indeed, this Special Issue boasts three new methodological approaches with Sheryl Homa and colleagues comparing two means of measuring oxidative stress (concluding that both used in tandem are better than one in isolation) [9] and both the Skinner and Champroux papers taking novel approaches to quantify the localization of chromosome territories in asymmetrical nuclei [5,6].…”