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* To whom correspondence should be addressed. E-mail: hallakj{at}androscience.com.br.
Although reactive oxygen species (ROS) are clearly implicated in the pathogenesis of male infertility, few studies have attempted to define the basal levels of ROS in fertile men. Levels of ROS are highly influenced by the presence of leukocytes and are associated with decreased seminal parameters. The objective of our study was to determine the normal ROS reference values in neat and washed semen of a fertile population and to correlate the leukocyte concentrations with seminal parameters. We evaluated 114 fertile men seeking vasectomy and 47 subfertile patients as a positive control. All samples were subjected to semen analysis and Endtz testing; chemiluminescence assay was used to determine ROS levels. All seminal parameters were significantly higher in the fertile men than in the subfertile patients. In non-leukocytospermic samples, ROS levels were lower in the fertile men than in the subfertile patients in neat (0.29 [0.18, 0.54] vs. 0.94 [0.38, 1.51]) (p=0.001) and washed semen (5.73 [1.90, 14.71] vs. 23.4 [9.46, 115.55]) (p=0.001). Similarly, in samples with leukocytes (Entdz<1x106/mL), ROS levels were lower in the fertile men in neat (0.75 [0.27, 1.71] vs. 2.0 [0.97, 27.41]) (p=0.001) and washed semen (15.85 [4.18, 62.16] vs. 239.83 [33.4, 1193.75]) (p<0.0001). As expected, samples with leukocytes had significantly higher ROS values in washed and neat semen. In the fertile population, ROS were positively correlated with leukocyte and negatively correlated with sperm count and motility. In semen samples without leukocytes, the normality cutoff of ROS was 0.55x104cpm with 76.4% AUC (area under the curve) in the neat samples and 10.0 with 77% AUC in the washed samples. In semen samples with leukocytes: the cutoff for ROS in neat samples was 1.25 with 72.7% AUC and 51.5 with 81% AUC in the washed samples. We defined the cutoff levels of ROS in a fertile population. Seminal leukocyte levels below 1x106/mL were associated with increased ROS. ROS levels were positively correlated with leukocytes and negatively correlated with sperm motility and concentration. Patients with normal seminal parameters and lower seminal leukocyte levels may benefit from therapeutic interventions that improve semen quality.
Key words: Endtz test •
male infertility •
oxidative stress •
spermatozoa •
standardization
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  K. Tremellen Oxidative stress and male infertility--a clinical perspective Hum. Reprod. Update, May 1, 2008; 14(3): 243 - 258. [Abstract] [Full Text] [PDF]
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