Abstract
The presence of high reactive oxygen species (ROS) levels in semen is a major factor involved in the decline of male fertility. In seminal plasma, ROS are mainly produced by activated leucocytes. Spermatozoa were the first cell type reported to show a potential susceptibility to oxidative damage. The aim of our study was to evaluate the impact of leucocytospermia on basal and FMLP (Formyl-Methionyl-Leucyl-Phenylalanine) induced oxidative status in semen of infertile men. We also analyzed the correlations of the spermatic parameters with amounts of ROS in semen. Our study included 50 semen samples of infertile men. Sperm analysis was performed using WHO standardized method. Seminal leucocytes were quantified using peroxidase technique. The measurement of ROS levels in semen was made by chemiluminescence assay. We measure respectively ROS amounts in neat semen and in washed sperm cells suspension from the same ejaculate. We also applied the test of provocation of leucocytes by FMLP on neat and washed samples to assess the spermatic oxidative status after leucocyte stimulation. Our results showed significant correlations between ROS levels in neat semen and many sperm parameters: motility, sperm concentration, leucocytes concentration and the rate of sperm cytoplasmic droplets. The studied samples were divided into 2 groups: (G1) composed of 36 samples without leucocytospermia and (G2) composed of 14 leucospermic samples. ROS levels were significantly lower in G1 than in G2 (p = 0.002). ROS production was significantly increased after application of FMLP in washed leucospermic samples (p = 0.001). The measurement of ROS in neat semen is a considerable contribution to explore the impairment of semen quality in infertile men. ROS levels in washed semen reflect the oxidative status generated by sperm preparation techniques used in assisted reproductive procedures. Levels of ROS are highly influenced by the presence of leucocytes and associated with decreased seminal parameters.
Highlights
Oxidative stress induced by reactive oxygen species (ROS) plays crucial roles in a wide range of physiological processes and is implicated in various diseases, such as cancer, cardiovascular pathology, neurodegenerative disorders, and other chronic conditions 1,2
We found significant and positive correlations of levels of basal ROS in neat semen with the leucocytes concentration in semen firstly and with the rates of cytoplasmic droplets in the mid piece area of spermatozoa secondly (Table 2; Figures 1(d) and (e))
It is well established that the production of excessive quantities of reactive oxygen species in the male genital tract, essentially by leucocytes, can overwhelm the seminal antioxidant defenses and give rise to a harmful oxidative stress altering the microenvironment in which spermatozoa develop and mature 5,18,27
Summary
Oxidative stress induced by reactive oxygen species (ROS) plays crucial roles in a wide range of physiological processes and is implicated in various diseases, such as cancer, cardiovascular pathology, neurodegenerative disorders, and other chronic conditions 1,2. The controlled generation of very low amounts of ROS appears to regulate sperm normal functions 3,4. Oxidative stress appears in semen once an imbalance between the production of ROS and their destruction by different enzymatic and non-enzymatic seminal antioxidant systems is created 5. Spermatozoa were the first cell type reported to show a potential susceptibility to oxidative damage 6. While the presence of high levels of ROS in the ejaculate is among the risk factors involved in reducing male fertility 7,8. It has evolved that three inter-related mechanisms account for OPEN ACCESS
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