Aitken R J, Clarkson J S
MRC Reproductive Biology Unit, Edinburgh, U.K.
J Reprod Fertil. 1987 Nov;81(2):459-69. doi: 10.1530/jrf.0.0810459.
Addition of the divalent cation ionophore, A23187, to washed populations of human spermatozoa resulted in a sudden burst of production of reactive oxygen species which peaked within 3-5 min. This activity was dependent upon the presence of calcium in the external medium and was unaffected by the mitochondrial inhibitors, oligomycin, antimycin and rotenone. Studies with scavengers of reactive oxygen species revealed that, while reagents directed against singlet oxygen and the hydroxyl radical were without effect, cytochrome C reduced the response to A23187 by about 50%, suggesting that the superoxide anion radical is a major product of the activated human spermatozoon. The clinical implications of these studies stem from the considerable variation observed between individuals in the levels of reactive oxygen species produced by the spermatozoa. This variability was shown to be inversely related to the ability of the spermatozoa to exhibit sperm-oocyte fusion on exposure to A23187; defective samples exhibited a basal level of reactive oxygen species production which was 40 times that observed with normal functional cells.
向洗涤过的人类精子群体中添加二价阳离子载体A23187，会导致活性氧的产生突然爆发，并在3 - 5分钟内达到峰值。这种活性依赖于细胞外培养基中钙的存在，并且不受线粒体抑制剂寡霉素、抗霉素和鱼藤酮的影响。对活性氧清除剂的研究表明，虽然针对单线态氧和羟基自由基的试剂没有效果，但细胞色素C使对A23187的反应降低了约50%，这表明超氧阴离子自由基是活化的人类精子的主要产物。这些研究的临床意义源于观察到的个体之间精子产生的活性氧水平存在相当大的差异。这种变异性被证明与精子在暴露于A23187时表现出精卵融合的能力呈负相关；有缺陷的样本表现出的活性氧产生基础水平是正常功能细胞的40倍。
