目的:探索在精子优化分离过程中,己酮可可碱(pentoxifylline,PF)和水溶性维生素E(water-soluble vitamin E,WsVitE)对改善精子质量的价值。方法:分别以密度梯度法、上泳法常用精子分离液作为对照组,在同批号对照组分离液中添加PF和/或WsVitE后作为试验组;将32份新鲜精液标本每份分为4份,分别以密度梯度法、上泳法2种方式优化分离,每种分离方式均设对照组和试验组;比较分离后精子的质量差异。结果:2种分离方法处理后的新鲜精子,其前向运动率、获能后酪氨酸磷酸化水平、Ca2+载体诱发精子顶体反应率,试验组均优于对照组(P<0.01);25℃孵育24 h后,试验组较对照组的精子前向运动率、活动指数、正常形态率更高,而DNA碎片、脂质过氧化水平更低(P<0.01);上泳法试验组分离率精子各项指标总体优于密度梯度法试验组(P<0.05)。结论:在精子分离液中加入抗氧化剂VitE和磷酸二酯酶抑制剂PF可明显改善分离后精子的质量;上泳法分离后的精子质量更高、损伤更轻。 Objective: To explore the value of pentoxifylline (PF) and water-soluble vitamin E (WsVitE) in improving sperm quality during sperm optimization. Methods: The density gradient method and the upper sperm method were respectively used as the control group. PF and / or WsVitE were added to the same batch of control group as the experimental group. 32 fresh semen samples were divided into 4 parts, Two methods of density gradient method and upper swimming method were used respectively to optimize the separation. Each method was divided into control group and experimental group. The difference of sperm quality was compared. Results: The sperm motility rate, tyrosine phosphorylation level and sperm acrosome reaction rate induced by Ca2 + carrier of fresh sperm treated by the two methods were all better than those of the control group (P <0.01). After incubation at 25 ℃ for 24 h, the sperm motility, activity index and normal morphology of the experimental group were higher than that of the control group, while DNA fragmentation and lipid peroxidation were lower (P <0.01) Rate of sperm indicators overall better than the density gradient test group (P <0.05). CONCLUSIONS: Antioxidant VitE and phosphodiesterase inhibitor PF can significantly improve the sperm quality after sperm separation. The sperm isolated by upper swimming method has higher quality and less damage.