Shekarriz M, Thomas A J, Agarwal A
Department of Urology, Cleveland Clinic Foundation, Ohio.
Urology. 1995 Jan;45(1):103-7. doi: 10.1016/s0090-4295(95)97088-6.
Excessive formation of reactive oxygen species (ROS) in human semen has been associated with impaired sperm function and fertility potential. The presence of ROS in semen specimens from normal fertile men emphasizes the importance of defining a normal range of ROS formation. The purpose of this study was to establish a normal range of ROS generation and to investigate the effect of sperm concentration on the ROS level.
ROS was determined in 15 healthy donors and 20 men with suspected infertility. After the sperm concentration in normal donors was adjusted to 20 x 10(6)/mL, ROS was measured by chemiluminescence using luminol in a Berthold luminometer. A specimen was regarded as positive (abnormal) when the value was at least 10 x 10(4) counts per minute (cpm). ROS was also evaluated at 4 sperm concentrations (60, 30, 15, and 7.5 x 10(6)/mL) from samples obtained from the patients with suspected infertility. In addition, ROS was measured in 7 ROS-positive specimens at a sperm concentration of 15 x 10(6)/mL and 60 x 10(6)/mL.
Results showed that ROS formation was negative in all 15 healthy donors (median, 0.9 x 10(4) cpm; interquartile range, 0 to 1.48 x 10(4) cpm). The ROS formation value among all the donors was less than 5.5 x 10(4) cpm. ROS formation was positive in 8 (40%) of the suspected infertile patients. ROS levels were significantly lower at sperm concentrations of 15 x 10(6)/mL or 7.5 x 10(6)/mL compared with 30 x 10(6)/mL or 60 x 10(6)/mL (P = 0.05). The ROS level increased after centrifugation for 10 minutes at 500 g in all 7 specimens at both 15 and 60 x 10(6)/mL. However, the increase in ROS formation at 60 x 10(6)/mL was significantly greater than that at 15 x 10(6)/mL (P < 0.001).
A range of ROS formation of 0 to 5.5 x 10(4) cpm at a sperm concentration of 20 x 10(6)/mL may be considered as normal for healthy donor semen. The positive relationship between ROS formation and sperm concentration at the time of measurement emphasizes the importance of concentration adjustment before analysis when comparing ROS levels between different specimens.
人类精液中活性氧（ROS）的过度生成与精子功能受损和生育潜能降低有关。正常可育男性精液标本中ROS的存在强调了确定ROS生成正常范围的重要性。本研究的目的是建立ROS生成的正常范围，并研究精子浓度对ROS水平的影响。
对15名健康供者和20名疑似不育男性的精液进行ROS测定。将正常供者的精子浓度调整至20×10⁶/mL后，使用鲁米诺在贝托尔德发光计中通过化学发光法测量ROS。当值至少为每分钟10×10⁴计数（cpm）时，标本被视为阳性（异常）。还对从疑似不育患者获得的样本在4种精子浓度（60、30、15和7.5×10⁶/mL）下评估ROS。此外，在7个ROS阳性标本中，分别在精子浓度为×10⁶/mL和60×10⁶/mL时测量ROS。
结果显示，所有15名健康供者的ROS生成均为阴性（中位数，0.9×10⁴ cpm；四分位间距，0至1.48×10⁴ cpm）。所有供者的ROS生成值均小于5.5×10⁴ cpm。8名（40%）疑似不育患者的ROS生成呈阳性。与30×10⁶/mL或60×10⁶/mL相比，精子浓度为15×10⁶/mL或7.5×10⁶/mL时ROS水平显著降低（P = 0.05）。在15×10⁶/mL和60×10⁶/mL的所有7个标本中，500 g离心10分钟后ROS水平均升高。然而，60×10⁶/mL时ROS生成的增加显著大于15×10⁶/mL时（P < 0.001）。
精子浓度为20×10⁶/mL时，ROS生成范围为0至5.5×10⁴ cpm可被视为健康供者精液的正常范围。测量时ROS生成与精子浓度之间的正相关关系强调了在比较不同标本的ROS水平时，分析前进行浓度调整的重要性。
