Villegas J, Kehr K, Soto L, Henkel R, Miska W, Sánchez R
Department of Medicine, Universidad de La Frontera, Temuco, Chile.
Andrologia. 2003 Aug;35(4):227-32. doi: 10.1046/j.1439-0272.2003.00564.x.
Leucocytospermia has been associated with reduced sperm motility and decreased capacity for sperm-egg interaction. This effect could be mediated through reactive oxygen species (ROS), which, at high concentrations, induce lipid peroxidation and cellular death. The high impact on sperm capacitation reported in other mammalians should be more accurately assessed in the human because premature activation could affect sperm fertilizing capacity. The aim of this study was to evaluate both the effect of ROS on sperm capacitation and the protective role of seminal plasma. Spermatozoa selected by Percoll gradient were incubated with polymorphonuclear (PMN) granulocytes isolated from blood and activated by phorbol-12-myristate 13-acetate (PMA). Different seminal plasma concentrations were added immediately or after 3-h incubation. Afterwards, ROS production was evaluated by luminescence and sperm capacitation by chlortetracycline stain. In PMN granulocytes and sperm suspensions, the basal ROS production was < 32 x 103 relative luminescence units (RLU). After stimulation with PMA, the rate of ROS production by PMN increased to 1,287 x 103 RLU. Incubation of sperm with activated PMN resulted in an increase of sperm capacitation (37% versus 19% in the control). Immediate addition of seminal plasma caused a significant reduction in ROS (P < 0.01) and prevented sperm from capacitating. A higher effect in inhibition of sperm capacitation was observed when seminal plasma had been added after 3-h incubation. The results suggest that human sperm capacitation can prematurely be induced by exogenous ROS and this effect can be reversed by seminal plasma. Thus, human sperm capacitation is another functional parameter that may be affected by nonphysiological ROS production.
白细胞精子症与精子活力降低及精卵相互作用能力下降有关。这种效应可能是通过活性氧（ROS）介导的，高浓度的ROS会诱导脂质过氧化和细胞死亡。其他哺乳动物中报道的ROS对精子获能的重大影响在人类中应得到更准确的评估，因为过早激活可能会影响精子的受精能力。本研究的目的是评估ROS对精子获能的影响以及精浆的保护作用。通过Percoll梯度选择的精子与从血液中分离并经佛波酯-12-肉豆蔻酸酯13-乙酸酯（PMA）激活的多形核（PMN）粒细胞一起孵育。立即或在孵育3小时后加入不同浓度精浆。之后，通过发光评估ROS的产生，并通过金霉素染色评估精子获能。在PMN粒细胞和精子悬液中，基础ROS产生量<32×103相对发光单位（RLU）。用PMA刺激后，PMN产生ROS的速率增加到1287×103 RLU。将精子与活化的PMN一起孵育导致精子获能增加（37% 对比对照组的19%）。立即加入精浆会使ROS显著降低（P<0.01）并阻止精子获能。在孵育3小时后加入精浆时，观察到对精子获能的抑制作用更强。结果表明外源性ROS可过早诱导人类精子获能，而这种效应可被精浆逆转。因此，人类精子获能是另一个可能受非生理性ROS产生影响的功能参数。
