冷冻干燥法对人精子DNA的影响  

Effect of freeze drying on the DNA of human spermatozoa

作　　者：沈大庆[1] 王沛涛[2] 李强[2] 刘忠强[2] 舒志全[3] 付秀艳[2] 李华琴[2] 

机构地区：[1]青岛大学医学院附属医院泌尿外科,266003 [2]青岛大学医学院附属医院低温医学科暨泌尿男科研究所,266003 [3]中国科学技术大学热科学与能源工程系,合肥230027

出　　处：《中国医药生物技术》2008年第1期49-53,共5页Chinese Medicinal Biotechnology

摘　　要：目的探讨冷冻干燥法用于人类精子保存的安全性。方法取健康志愿者合格精液40份,平均分为4组,其中3组分别加入不同的冻干保护剂(ETBS;ETBS+海藻糖;ETBS+海藻糖+蛋黄)后给予冷冻干燥处理,在4℃冰箱中保存3周;1组作为新鲜精液对照组。对4组标本分别以原位缺口末端标记(TUNEL)法和彗星试验进行DNA断裂精子百分率检测。结果ETBS、ETBS+海藻糖、ETBS+海藻糖+蛋黄组和新鲜精液组DNA断裂精子百分率以TUNEL法检测,分别为(6.39±1.46)%、(5.75±1.29)%、(5.20±1.38)%、(4.94±1.86)%;以彗星试验检测,分别为(6.48±1.58)%、(5.83±1.48)%、(5.28±1.42)%、(5.12±1.65)%。冷冻干燥保存后的各组与新鲜精液相比较,DNA断裂精子百分率差异均无统计学意义(P>0.05)。结论以ETBS或ETBS加海藻糖、蛋黄为保护剂的冷冻干燥法对人精子DNA无明显损伤,可有效地保护人精子的DNA。Objective To explore the security of freeze drying in human semen preservation. Methods Forty qualified semen samples were obtained from healthy volunteers, and divided equally into 4 groups. Three semen sample groups were added with freeze drying protective agents, ETBS, ETBS ＋ trehalose, or ETBS ＋ trehalose ＋ egg yolk, respectively, and then conserved in a refrigerator at 4℃ for 3 weeks. Another group of semen sample was used as a fresh sperm control. In situ nick-end labeling （TUNEL） and comet assay were used to detect the sperm DNA damage. Results The percentage of TUNEL-positive spermatozoa in the ETBS, ETBS ＋ trehalose, ETBS ＋ trehalose ＋ egg yolk, and control groups were （6.39± 1.46）%, （5.75±1.29）%, （5.20±1.38）%, and （4.94±1.86）%, respectively; while these detected by Comet assay were （6.48 ±1.58）%, （5.83±1.48）%, （5.28±1.42）%, and （5.12±1.65）%, respectively. No significant difference was found in the percentage of spermatozoa with damaged DNA between fresh and freeze dried spermatozoa （P〉0.05）. Conclusion Freeze drying using ETBS, ETBS ＋ trehalose, or ETBS ＋ trehalose ＋ egg yolk have no damage to the DNA of human spermatozoa, which can protect the DNA of human spermatozoa effectively.

关 键 词：冷冻干燥法 精液保存 原位缺口末端标记 彗星试验 

分 类 号：R318.52[医药卫生—生物医学工程] R321[医药卫生—基础医学]