三种精子质膜完整性检测方法的比较  

Comparative study of three methods for evaluating membrane integrity of human sperm

作　　者：舒如明[1,2] 卢文红[2] 李鸿[2] 周芳[2] 许剑锋[2] 王小强[1] 谷翊群[2] 

机构地区：[1]北京协和医学院研究生院,北京100730 [2]国家人口计生委科学技术研究所男性临床研究室,北京100081

出　　处：《生殖医学杂志》2009年第6期535-539,共5页Journal of Reproductive Medicine

基　　金：中央级公益性科研所基本科研业务费专项(2007计生所JZ01)

摘　　要：目的对6-羧基二乙酸荧光素／碘化丙啶（6-CFDA／PI）双荧光标记方法进行改良并应用于人精子质膜完整性的检测，并与单用伊红Y染色、低渗肿胀试验（HOS）两种检测方法进行比较。方法随机选择30例本所人类精子库志愿者精液标本（密度≥20×10^6／ml、活力（a＋b）≥50%），分别采用单用伊红Y染色、HOS和6-CFDA／PI双荧光标记试验检测精子质膜完整性。结果30例精液标本伊红Y染色活精子比率、HOS肿胀精子数比率、6-CFDA／PI双荧光标记结果分别为（68．8±7．6）％、（68．7±7．8）％和（67．9±7．8）％，两两比较，无显著性差异（P均〉0．05），但双荧光标记能显示死活精子的过渡状态。相关性分析结果显示，三种方法检测的活精子比率均与前向运动（a＋b）精子呈显著正相关。结论6-CFDA／PI双荧光标记检测方法与单用伊红染色、HOS两种常用检测精子质膜完整性方法具有一致性，同时能鉴定出精子过渡的中间状态。Objective： To modify a method for clinical application to assess membrane integrity of human sperm by means of dual fluorescent staining technique using 6-carboxyfluorescein diacetate （6-CFDA）and propidium iodide（PI）in comparison with the techniques of single Eosin Y staining test or hypo-osmotic swelling test. Methods： Semen specimen（concentration≥20 × 10^6/ml, motility a＋ b≥50 %） was randomly obtained from 30 healthy volunteers in human sperm bank of National Research Institute for Family Planning. Membrane integrity of human sperm was evaluated by single Eosin Y staining, hypo osmotic swelling and dual fluorescent staining technique, respectively. Results： The rates of live sperm detected by single Eosin Y staining, hypo-osmotic swelling and dual fluorescent staining technique were（68.8± 7. 6）%, （68.7 ± 7.8） % and（67. 9 ± 7.8） %, respectively（P〉 0.05） while dual fluorescent staining technique using 6-CFDA and PI could demonstrate the transition from live sperm to dead sperm. Correlation analysis showed that progressive motility（a＋b）of sperm was significantly correlated with the rates of live sperm detected by three methods. Conclusions： Dual fluorescent staining technique using 6 CFDA and PI could be used to assess membrane integrity of human sperm and the result is consistent with the single Eosin Y staining or hypo-osmotic swelling. In addition, it is characterized by evaluating intermediate state between live sperm and dead sperm.

关 键 词：完整性评价 精子膜 人类 评估 服装设计师 染色技术 染色试验 曙红Y 

分 类 号：TS941.2[轻工技术与工程—服装设计与工程] TD4[矿业工程—矿山机电]