Evaluation for the effect of seminal plasma reactive oxygen species on infertility during sperm dna fragmentation and apoptosis

The source for spermatozoon DNA damage and it?s significance in infertility have been explained with various mechanisms by researchers so far. The emphasized subjects were protamination, apoptosis and oxidative stress. While there are papers submitting that smoking affects the semen quality in human, there are also papers which establish no significant relation between the parameters of smoking and spermatozoon. On this study, we intend to evaluate the effect of seminal plasma reactive oxygen species on infertility during sperm DNA fragmentation and apoptosis.Totally 50 cases were evaluated in the scope of the project. Semen samples were analyzed from twenty seven patients with asthenozoospermia and twenty three patients with normozoospermia (Smoking and non-smoking). Sperm concentration and motility rates were evaluated microscopically in accordance with World Health Organization (WHO) criterias while its morphology was assessed according to the Kruger?s strict criterias by using STAT III stain. For vitality, Eosin Y assay applied to the semen samples and assessed with light microscopy. Semen samples were separated to three groups after each case. TUNEL assay was used to detect of DNA fragmentation, MDA assessment for ROS, GPX for antioxidant enzyme respectively. Annexin V assay used to determine apoptosis in TUNEL positive cases when apoptosis rate was >%10. Apoptosis was evaluated ultrastructurally by transmission electron microscopy.We saw that there is no significantly difference between smoking and semen parameters (concentration, motility, morphology) in cases with normozoospermia and asthenozoospermia . There was difference DNA fragmentation between asthenozoospermia and control group (normozoospermia). A negative correlation was found between DNA fragmentation and semen parameters, vitality. A significantly difference was found between the levels of MDA and GPX in two groups. MDA was no significant in spite of difference between smoking and non smoking groups. There was no significant difference between ROS and semen parameters, vitality in two groups. A positive correlation was found between ROS and DNA fragmentation. A negative correlation was found between ROS and GPX. These findings were supported by ultrastructurally.Consequently, on the contrary with papers claiming that ROS levels increased in smokers induce apoptosis and so directly affect the infertility, in this paper, the effect of smoking on ROS levels is not found statistically significant and so its effect on apoptosis and infertility is not established.