摘要
目的 观察密度梯度离心法联合上游法优选精子前后各项参数的变化情况,证实优选技术对精液参数的改善作用,探讨它们之间的内在联系.方法 密度梯度离心法联合上游法精子优选前后,使用计算机辅助精子活动分析仪检测精子活动力变化;使用流式细胞仪,检测早期凋亡、MMP和细胞内ROS;使用单细胞凝胶电泳观察nDNA完整性变化.结果 优选后精子运动能力、正常形态率和nDNA完整性明显提高,差异具有统计学意义;优选前,MMP与a级精子百分比、VSL具有正相关（相关系数分别为0.641,0.708,P＜0.05）;当白细胞≤1 × 10^6/ml时,优选前和优选后正常形态率与ROS呈负相关（相关系数分别为-0.629,-0.685,P＜0.05）.结论 密度梯度离心法联合上游法优选精子是安全有效的方法;MMP能反映精子的运动能力;形态不正常的精子是ROS的重要来源.
Objective To observe sperm changes before and after the preparation of the density gradient centrifugation and swim-up,to confirm the role of the preferred technology in improvement the quality of the sperm,and to explore internal relations between them.Methods Semen samples were prepared by sperm prepared technology （the density gradient centrifugation and swim-up）.Then early apoptosis,mitochondrial membrane potential,reactive oxygen specials were evaluated by flow cytometry analysis.Sperm parameters were examined by computer assisted sperm analysis.The nDNA integrity was evaluated by the single cell gel electrophoresis.Results Sperm motility and the percentage morphologically normal spermatozoa improved significantly after the density gradient centrifugation and swim-up compared with raw semen.MMP were significantly correlated with the percentage a sperm and VSL （r=0.641,0.708,P ＜ 0.05）.ROS had the negative correlation with the percentage morphologically normal spermatozoa in non-leucocytospermia patients （r=-0.629,-0.685,P ＜ 0.05）.Conclusion The method of density gradient centrifugation and swim-up was safe and effective; MMP reflect sperm motility; morphologically unnormal spermatozoa sperm is an important source of ROS in non-leucocytospermia patients.
出处
《国际医药卫生导报》
2014年第9期1221-1224,共4页
International Medicine and Health Guidance News
关键词
精子
优选
凋亡
线粒体膜电位
活性氧
单细胞凝胶电泳
Spermatozoa
Apoptosis
Mitochondrial membrane potential
Reactive oxygen specials
Single cell gel electrophoresis
