de Lamirande E, Gagnon C
Urology Research Laboratory, Royal Victoria Hospital, Montreal, Quebec, Canada.
J Androl. 1992 Sep-Oct;13(5):379-86.
Under moderate conditions, reactive oxygen species (ROS) have been shown to inhibit sperm motility after several hours of incubation. The rapid decrease in flagellar beat frequency observed within the first hour of contact between ROS and spermatozoa was associated with a rapid loss of intracellular adenosine triphosphate (ATP). Motility of intact spermatozoa ceased when their ATP concentration was reduced by 85 +/- 5%. Axonemal damage was confirmed when ROS-treated spermatozoa could not reactivate motility after demembranation in a medium containing magnesium adenosine triphosphate (Mg.ATP). However, in conditions allowing rephosphorylation of the axonemes (addition of cyclic adenosine monophosphate, or cAMP, and protein kinase or sperm extracts to the demembranation medium), the motility could reactivate. Three lines of evidence suggested that ATP depletion induced by ROS treatment was responsible for the effects observed in spermatozoa. First, the rapid decrease in intracellular ATP observed after ROS treatment was closely followed by a decrease in beat frequency, loss of intact sperm motility, and axonemal damage due to insufficient phosphorylation. Second, incubation of spermatozoa with the combination pyruvate-lactate allowed maintenance of sperm ATP at a normal level and prevented the effects of ROS; furthermore, spermatozoa immobilized after ROS treatment, then supplemented with pyruvate-lactate, were able to reinitiate motility in parallel with an increase in their ATP level. Third, treatment of spermatozoa with rotenone, an ATP depleting agent, produced effects similar to ROS treatment and could also be reversed by the addition of pyruvate-lactate. These data are consistent with the conclusion that ROS treatment produced axonemal damage mostly as a result of ATP depletion.(ABSTRACT TRUNCATED AT 250 WORDS)
在适度条件下，活性氧（ROS）已被证明在孵育数小时后会抑制精子活力。在ROS与精子接触的第一小时内观察到的鞭毛摆动频率的迅速下降与细胞内三磷酸腺苷（ATP）的快速损失有关。当完整精子的ATP浓度降低85±5%时，其活力停止。当用ROS处理的精子在含有镁三磷酸腺苷（Mg.ATP）的介质中去膜后无法重新激活活力时，轴丝损伤得到证实。然而，在允许轴丝重新磷酸化的条件下（向去膜介质中添加环磷酸腺苷，或cAMP，以及蛋白激酶或精子提取物），活力可以重新激活。三条证据表明，ROS处理诱导的ATP消耗是精子中观察到的这些效应的原因。首先，ROS处理后观察到的细胞内ATP的快速下降紧接着是摆动频率的下降、完整精子活力的丧失以及由于磷酸化不足导致的轴丝损伤。其次，将精子与丙酮酸 - 乳酸组合孵育可使精子ATP维持在正常水平，并防止ROS的影响；此外，用ROS处理后固定的精子，再补充丙酮酸 - 乳酸，能够随着ATP水平的增加重新启动活力。第三，用鱼藤酮（一种ATP消耗剂）处理精子产生的效应与ROS处理相似，并且也可以通过添加丙酮酸 - 乳酸来逆转。这些数据与以下结论一致，即ROS处理主要由于ATP消耗而导致轴丝损伤。（摘要截短于250字）
