Human sperm have been documented to display apoptosis-like features such as externalization of phosphatidylserine (EPS), disruption of the transmembrane mitochondrial potential (MMP) and activation of caspases. Our aim was to evaluate possible association between activation of the apoptosis cascade in human sperm and its oocyte penetration capacity using the zona free hamster oocyte penetration assay (SPA). Semen specimens from 76 unselected donors were subjected to double density gradient centrifugation followed by incubation under capacitating conditions for 3 h and SPA. Apoptosis signalling was monitored by assessment of EPS, disruption of MMP and activation of caspase-3 by flow cytometry. Semen samples with subnormal SPA values (<20% penetrated oocytes) contained significantly higher amounts of spermatozoa with EPS, disrupted MMP and activated caspase-3 compared with those samples with normal SPA values (>20% penetrated oocytes, p < 0.01). All three apoptosis markers showed a significantly negative correlation with the percentage of penetrated oocytes (p < 0.01). Apoptosis-related signalling appears to have a negative association with sperm-oocyte penetration. The exclusion of sperm presenting with those apoptosis-related features during assisted reproduction may improve success rates of procedures such as intrauterine insemination and in vitro fertilization.

译文

人类精子已被证明具有凋亡样特征,例如磷脂酰丝氨酸 (EPS) 的外部化,跨膜线粒体电位 (MMP) 的破坏和胱天蛋白酶的激活。我们的目的是使用无带仓鼠卵母细胞渗透试验 (SPA) 评估人类精子中凋亡级联的激活与其卵母细胞渗透能力之间的可能关联。对来自76个未选择的供体的精液标本进行双密度梯度离心,然后在容量条件下孵育3小时和SPA。通过流式细胞术评估EPS,MMP破坏和caspase-3激活来监测凋亡信号。与具有正常SPA值 (>20% 穿透卵母细胞,p <0.01) 的那些样品相比,具有低于正常SPA值 (<20% 穿透卵母细胞) 的精液样品含有显著更高量的具有EPS、破坏的MMP和活化caspase-3的精子。3种凋亡标记物均与穿透卵母细胞的百分比呈显著负相关 (p <0.01)。凋亡相关信号似乎与精子卵母细胞的渗透呈负相关。在辅助生殖过程中排除具有凋亡相关特征的精子可能会提高子宫内人工授精和体外受精等手术的成功率。

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