马春杰 庄嘉明 邓顺美 唐运革 秦卫兵 王奇玲 张欣宗

[摘要]目的 研究甘油-卵黃-柠檬酸盐（GEYC）冷冻保护剂中添加不同摩尔浓度二十二碳六烯酸（DHA）对人精子冷冻前后运动学参数的影响，探索n-3多不饱和脂肪酸在人精子冷冻中的应用价值。方法 选取2016年3月～2017年4月广东省计划生育专科医院人类精子库的筛查期供精者的33例精液标本，以GEYC冷冻保护剂为基础，外源性DHA在精液-GEYC混合液中的终浓度分别为对照组0 mmol/L、DHA 1组0.317 mmol/L、DHA 2组0.634 mmol/L、DHA 3组1.268 mmol/L。采用SCA 5.2计算机辅助精子分析系统分别检测添加冷冻保护剂后以及冷冻复苏后0～8 h的精子运动学参数。结果 冷冻前精液加入各组保护剂后，仅DHA 3组的VCL和VAP显著低于对照组（P<0.05），其他各组间的各项运动学参数差异均无统计学意义（P>0.05）；冷冻复苏后0、4、5、6、8 h，仅见DHA 3组偶有运动学参数下降，与其他3组比较差异有统计学意义（P<0.05），冻后0～8 h大部分运动学参数各组间差异均无统计学意义（P>0.05）。结论 甘油-卵黄-柠檬酸盐冷冻保护剂中添加DHA未见冷冻前后人精子运动学参数的提高，适量添加DHA可能通过其他途径对冷冻精子产生影响，大量添加DHA可能对冷冻前后部分运动学参数有负面影响。

[关键词]n-3多不饱和脂肪酸；精液冷冻；精子运动学参数；二十二碳六烯酸

[中图分类号] R691.9 [文献标识码] A [文章编号] 1674-4721（2018）4（a）-0004-05

Influence of addition of different molar concentration of DHA into Glycerol-Egg Yolk-Citrate Cryopreservative Mediums on the kinematic parameters of human sperm before and after cryosurgery

MA Chun-jie1 ZHUANG Jia-ming1 DENG Shun-mei1 TANG Yun-ge2 QIN Wei-bing1，2 WANG Qi-ling1 ZHANG Xin-zong1

1.Department of Andriatrics，Guangdong Institute of Family Planning Science and Technology （Guangzhou Family Planning Specialty Hospital），Guangzhou 510600，China；2.Key Laboratory of Male Reproductive and Heredity，State Health and Family Planning Commission，Guangdong Institute of Family Planning Science and Technology，Guangzhou 510600，China

[Abstract]Objective To study the influence of addition of different molar concentration of docosahexaenoic acid （DHA） into Glycerol-Egg Yolk-Citrate （GEYC） cryopreservative mediums on the kinematic parameters of human sperm before and after cryosurgery so as to explore the application value of n-3 polyunsaturated fatty acids in cryosurgery of human sperm.Methods Altogether 33 cases of semen samples which came from semen donors during human sperm bank screening period in Guangzhou Family Planning Specialty Hospital from March 2016 to April 2017 were selected.Based on GEYC cryopreservative mediums，the concentration of exogenous DHA in mixed semen-GEYC liquor were 0 mmol/L in the control group，0.317 mmol/L in the DHA 1 group，0.634 mmol/L in the DHA 2 group and 1.268 mmol/L in the DHA 3 group.SCA 5.2 computer-assisted sperm analysis system was used to detect the sperm kinematics parameters 0-8 hours after addition of cryopreservation and frozen resuscitation.Results After addition of cryopreservation and frozen resuscitation，only curve line velocity （VCL） and average path velocity （VAP） of the DHA 3 group were significantly lower than those of the control group （P<0.05），there was no statistically significant difference in kinematic parameters between the other groups （P>0.05）.After frozen recovery at 0，4，5，6，8 hours，only kinematic parameters of the DHA 3 group decreased occasionally，with statistically significant difference in contrast with 3 groups.After cryosurgery at 0-8 hours，most of the kinematic parameters had no statistically significant difference between each group （P>0.05）.Conclusion Addition of DHA into GEYC cryo-preservative mediums do not increase the sperm kinematic parameters before and after cryosurgery.The proper addition of DHA may affect frozen semen through other ways.Addition of a large number of DHA may have a negative impact on the part of the kinematic parameters before and after cryosurgery.

[Key words]n-3 polyunsaturated fatty acids；Semen cryosurgery；Sperm kinematics parameters；Docosahexaenoic acid

精液的冷冻保存是精子库和辅助生殖技术实验室的一项重要工作。冷冻复温过程能够破坏人精子结构的完整性，从而影响精子功能。很多动物实验显示，饮食添加[1-6]和冷冻稀释液[7]中直接添加n-3多不饱和脂肪酸（n-3 polyunsaturated fatty acids，n-3 PUFA），尤其是添加二十二碳六烯酸（docosahexaenoic acid，DHA）（n-3 PUFA的一种）可以提高哺乳动物新鲜和冷冻后精液质量。Vireque等[8]的研究显示，含有卵黄的人精子冷冻保护剂中添加富含n-6和n-3 PUFA的大豆卵磷脂可以使人精子有效抵御冷冻损伤。……