Agarwal A, Ikemoto I, Loughlin K R
Division of Urology, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts.
Arch Androl. 1994 Nov-Dec;33(3):157-62. doi: 10.3109/01485019408987819.
The possibility was evaluated that the production of reactive oxygen species (ROS) by human sperm is stimulated by the repeated cycles of centrifugation and resuspension involved in conventional sperm preparation. ROS generation by human sperm was monitored before and after the washing of sperm from 55 men (43 men with suspected subfertility and 12 normal volunteers). The ROS activity of all 55 specimens before washing was inversely correlated with original sperm motility (r = .278, p < .05). The mean level of ROS activity was significantly higher after washing than before processing (p < .05) for the 26 specimens with normal sperm motility, the 20 specimens with normal sperm morphology, and the 12 specimens with both normal motility and normal morphology. In contrast, the mean ROS level was not significantly changed after washing in the 27 specimens with poor sperm motility, the 16 specimens with abnormal sperm morphology, or the 13 specimens with both abnormal motility and abnormal morphology. It would appear that repeated centrifugation, resuspension, and vortexing cause excessive generation of ROS in the motile sperm population of the washed specimen. Washing procedures involving excessive manipulation of sperm may, in fact, cause the most harm to motile sperm, i.e., those that the method is trying to select. Procedures that minimize multiple centrifugation, resuspension, and vortexing steps should therefore be used for the preparation of semen specimens for assisted-reproduction techniques.
评估了常规精子制备过程中涉及的反复离心和重悬操作是否会刺激人类精子产生活性氧（ROS）。监测了55名男性（43名疑似生育力低下男性和12名正常志愿者）精子洗涤前后的ROS生成情况。所有55个样本洗涤前的ROS活性与原始精子活力呈负相关（r = 0.278，p < 0.05）。对于精子活力正常的26个样本、精子形态正常的20个样本以及精子活力和形态均正常的12个样本，洗涤后的ROS活性平均水平显著高于处理前（p < 0.05）。相比之下，精子活力差的27个样本、精子形态异常的16个样本或精子活力和形态均异常的13个样本，洗涤后的ROS平均水平没有显著变化。似乎反复离心、重悬和涡旋会导致洗涤后样本中有活力精子群体过度产生ROS。实际上，涉及对精子过度操作的洗涤程序可能对活力精子造成最大伤害，即该方法试图挑选的精子。因此，在为辅助生殖技术制备精液样本时，应采用尽量减少多次离心、重悬和涡旋步骤的程序。
