国际生殖健康/计划生育杂志

国际生殖健康/计划生育 ›› 2016, Vol. 35 ›› Issue (5): 369-371.

• 论著 • 上一篇    下一篇

体外操作对人精子运动参数及细胞内活性氧簇的影响

史潇，王婷，周瑶，刘厉，詹晓敏，陈思梅，全松   

  1. 510515 广州，南方医科大学南方医院生殖中心
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2016-10-25 发布日期:2016-10-25
  • 通讯作者: 全松

The Effect of Physical Stress on Human Sperm Motility and Intracellular ROS Generation

SHI Xiao，WANG Ting，ZHOU Yao，LIU Li，ZHAN Xiao-min，CHEN Si-mei，QUAN Song   

  1. Center for Reproductive Medicine，Nanfang Hospital，Southern Medical University，Guangzhou 501515，China
  • Received:1900-01-01 Revised:1900-01-01 Published:2016-10-25 Online:2016-10-25
  • Contact: QUAN Song

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摘要： 目的：研究体外物理操作（离心以及微量加样器抽吸）对人精子运动参数以及细胞内活性氧簇（ROS）的影响，旨在优化人精子体外处理方法。方法：7例精液常规参数正常的标本，采用不同离心力（200 g，600 g）及离心时间（5 min，15 min）以及不同微量加样器抽吸次数（2，6，10次）进行处理，测试精子运动参数和细胞内ROS的变化。结果：①活动精子百分比以及活精子细胞内ROS水平主要受到离心时间的影响（P＜0.05）；前向运动精子百分比（PR）以及平均运动速率（VAP）均不受离心力及离心时间的影响（P＞0.05）。单因素方差分析多重比较结果提示，当离心时间为5 min以上时精子活动力显著下降，而ROS水平显著增加（P＜0.05）。②加样器抽吸2次以上可致人精子活动力显著下降（P＜0.05），抽吸6次以上可导致人精子PR及VAP显著降低（P＜0.05）；但抽吸操作并未显著影响精子细胞内ROS的水平。结论：在人精子体外操作时应尽量缩短离心时间并控制加样器抽吸次数从而保证正常的精子运动参数，也应尽量缩短离心时间从而减少体外操作所致的细胞内ROS升高。

关键词: 精子, 精子能动性, 离心法, 梯密度, 抽吸, 氧化性应激, 受精, 体外

Abstract: Objective: To study the effects of centrifugation and pipette on human motility and intracellular ROS, so as to optimize the sperm handling procedure. Methods：Seven sperm samples with normal seminal parameters were treated with various centrifugation force （200 g，600 g) and centrifugation duration (5 min，15 min） and different pipetting times （2，6，10 times）. Motility parameter and intracellular ROS were measured. Results：For the centrifuge procedure, the centrifugation duration could significantly influence the sperm motility and intercellular ROS level （P＜0.05）, while centrifugation did not significantly influence the percentage of progressive motile sperm （ＰＲ） and the velocity of average path（VAP）（P＞0.05）. The centrifugation duration more than 5 minutes could significantly decrease the sperm motility and intracellular ROS level（P＜0.05）. For the pipetting procedure, the pipetting more than 2 times would significantly decrease the sperm motility （P＜0.05），while the pipetting over 6 times would significantly decrease the PR and VAP （P＜0.05）. However, the intracellular ROS level was not significantly changed by the repeated pipetting. Conclusions：The special causion is that the centrifuge over 5 minutes and pipetting over 2 times should be avoided during in vitro handling human sperm sample.

Key words: Spermatozoa, Sperm motility, Centrifugation, density gradient, Suction, Oxidative stress, Fertilization in vitro