摘要
目的观察优选技术在精液冷冻保存中对常规精液参数、线粒体膜电位(△Ψm)和活性氧(ROS)的影响。方法分别用计算机辅助精子分析仪(CASA)、流式细胞仪(FCM)检测优选或未优选的精子在冷冻前后常规精液参数、△Ψm和精子细胞内ROS的变化。结果各冻融组精子活率、a级精子、(a+b)级精子、正常形态率均比冷冻前有不同程度降低,有显著性差异;而优选冻融组和优选加精浆冻融组间常规精液参数无显著性差异(P>0.05),但都明显高于原始冻融组(P<0.01)。原始精液冷冻后比冷冻前△Ψm下降(P<0.05),ROS显著上升(P<0.01)。优选冻融组和优选加精浆冻融组与冷冻前相比,以及组间无显著性差异(P>0.05);但与原始冻融组分别比较,△Ψm均显著提高(P<0.05),ROS均显著减少(P<0.01)。各冻融组△Ψm和ROS都有明显相关性,相关系数(r)依次为-0.528、-0.537、-0.606(P<0.001)。结论精液优选后冷冻可以在保持良好的△Ψm同时,减少ROS生成,因此建议临床可应用该技术冷冻保存精子。
Objective： To investigate effects of cryopreservation and preparation on sperm parameters, mitochondrial membrane potential （△ψm） and reactive oxygen species （ROS）.
Methods： The △ψm, ROS and parameters of sperm samples （raw semen, prepared sperm, frozenthawed raw semen, frozen-thawed prepared sperm and frozen-thawed prepared sperm in plasma） were analyzed by flow cytometry （FCM） and computer-assisted semen analysis（CASA）.
Results： The sperm motility, viability and percentage of sperm with normal morphology of the frozenthawed samples decreased significantly, while the significant differences were not found between the frozen-thawed prepared sperm and thawed prepared sperm in plasma （P〉0.05）. The sperm parameters of the frozen-thawed prepared samples whether in plasma or not increased significantly compared to the frozen-thawed raw semen （P〈0.01）. Compared to that of raw semen, the i^m of frozen-thawed sperm decreased significantly （P〈0.05）, while the ROS increased significantly （P〈0.01）. The significant differ ences were not found among the frozen-thawed prepared sperm, frozen-thawed prepared sperm in plasma and prepared sperm （P〉0.05）. The △ψm of frozen-thawed prepared sperm whether in plasma or not increased significantly （P〈0.05） and ROS of the same groups decreased significantly （P〈0.01） when compared with that of frozen-thawed raw semen. In groups of frozen-thawed raw sperm, frozen-thawed pre- pared sperm and frozen-thawed prepared sperm in plasma, the correlation coefficients （r） of △ψm and ROS were -0. 528, -0. 537 and -0. 606 respectively. The differences were significant （P〈0.001）.
Conclusions. Cryopreservation of prepared sperm can significantly decrease ROS production and keep high △ψm. We recommend this technology for the cryopreservation of human sperm.
出处
《生殖医学杂志》
CAS
2006年第4期237-240,共4页
Journal of Reproductive Medicine
基金
广东省科技厅项目(2005B34201013)
汕头市科技厅项目(200525)
关键词
精液
冷冻
线粒体膜电位
活性氧
常规精液参数
流式细胞术
Human sperm
Cryopreservation
Mitochondrial membrane potentiaL
Reactive oxygen species
Sperm parameters
Flow cytometry
