精液保存和优化处理方法对精子DNA完整性的影响  

Influence of semen preservation and processing methods on sperm DNA integrity

作　　者：郑毅春[1] 梁嘉颖[1] 杜鹏[1] 莫国柱[1] 汪李虎[1] 刘风华[1] 

机构地区：[1]广东省妇幼保健院生殖健康与不孕症科,广东广州510010

出　　处：《中华男科学杂志》2016年第5期432-436,共5页National Journal of Andrology

摘　　要：目的：探讨不同精液保存和处理方法对精子DNA完整性的影响。方法：筛选2015年1~12月就诊的100例精液正常志愿者,将每例志愿者的精液混匀后分别对其进行不同的保存和优化处理;保存方法包括直接冷冻法、试剂冷冻法、室温保存4 h和24 h等,优化处理方法包括简单洗涤法、直接上游法和非连续密度梯度离心法。保存和处理后均采用精子染色体扩散实验（SCD）分析其精子DNA碎片指数（DFI）;优化处理各组继续培养24 h,培养后再分析其精子活动率和DFI。结果：精液保存条件两两组间分析显示,直接冷冻法、试剂冷冻法和室温保存4 h的精子DFI分别为（27.3±6.4）%、（26.9±6.1）%和（24.7±6.8）%,结果无显著性差异（P〉0.05）,而室温保存24 h则显著增加精子DFI[（35.6±9.0）%]（P〈0.05）。与简单洗涤法的精子DFI[（13.7±2.0）%]相比,直接上游法的精子DFI[（9.1±1.3）%]和非连续密度梯度离心法的精子DFI[（8.0±2.5）%]均显著降低（P〈0.05）;再培养24 h后,非连续密度梯度离心法处理的精子DFI[（11.5±4.2）%]最低（P〈0.05）。结论：保存条件和优化处理方法对精子DNA完整性有影响,临床工作中需选取最合适的精液处理方法。Objective： To investigate the influence of different methods of semen preservation and processing on sperm DNA integrity. Methods： We collected semen samples from 100 normozoospermic male volunteers and,following homogeneous mixing,preserved them by means of snap freezing,slow freezing,or at the room temperature for 4 and 24 hours. Meanwhile we processed the semen by washing,swim-up,and density gradient centrifugation（ DGC）. Then we obtained the sperm DNA fragmentation index（ DFI）by sperm chromatin dispersion test and measured total sperm motility and DFI after cultured for 24 hours following processing. Results： The sperm DFIs after 4 hours of preservation by snap freezing,slow freezing,and at the room temperature were（ 27. 3 ±6. 4） %,（ 26. 9 ± 6. 1） %,and（ 24. 7 ± 6. 8） %,respectively,and that after preserved at the room temperature for 24 hours was（ 35. 6 ± 9. 0） %,with statistically significant differences between the first three and the 24-hour room temperature preservation groups（ P〈0. 05） but not among the former three groups（ P〉0. 05）. The sperm DFI was significantly higher in the samples processed by washing（ [13. 7 ± 2. 0]%） than in those processed by swim-up（ [9. 1 ± 1. 3]%） and DGC（ [8. 0 ± 2. 5]%）（ P〈0. 05）,and it was the lowest in the DGC group after 24-hour culture（ [11. 5 ± 4. 2]%） as compared with the other groups（ P〈0. 05）. Conclusion： Sperm DNA integrity is influenced by different semen preservation conditions and processing methods.

关 键 词：精液保存 精液优化处理 DNA碎片 精子染色体扩散实验 

分 类 号：R698.2[医药卫生—泌尿科学]