己酮可可碱和水溶性维生素E在精子优选过程中对精子质量的影响

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。

Effects of Adding Pentoxifylline and Water-soluble Vitamin E into Sperm Preparation Media on Selected Spermatozoa Qualities

Objective: To explore the value of sperm separation from seminal plasma by adding pentoxifylline(PF) and water-soluble vitamin E(WsVitE) into sperm preparation media for improving sperm quality.Methods: WsVitE and PF were added into traditional culture media which were employed to select sperm by gradient-density centrifugation and swim-up preparation techniques.Traditional culture media of same lot number was defined to the control.Traditional culture media(control group) and modified media containing WsVitE and PF additives(test group) were applied to separate sperm from 32 fresh semen samples by gradient-density centrifugation and swimup methods synchronically.The quality of prepared sperm fractions was compared between the test group and the control,and the advantage of sperm preparation techniques was assessed between gradient-density centrifugation and swim-up.Results: In the final sperm preparation of both gradient-density centrifugation and swim-up,the proportions of progressive motility(PM),the abilities of capacitated sperm protein tyrosine phosphorylation(TP) and acrosome reaction after ionophore challenge(ARIC) in test group were more excellent than those in the control(P<0.01).After selected sperm was incubated for 24 h at 25℃,the proportions of PM,index of motility,the rate of normal morphology were higher,and the levels of DNA fragmentation and lipid peroxidation(LPO) were lower in test group than in the control(P<0.01).Furthermore,the characteristics of selected sperm by swim-up were more perfect than those by gradient-density centrifugation(P<0.05).Conclusion: Sperm quality can be improved by adding antioxidant WsVitE and phosphodiesterase inhibitor PF into culture media.Swim-up preparation technique is more advantageous than gradient-density centrifugation in enhancing sperm quality and decreasing damage.