彗星实验在不明原因复发性流产中对精子DNA完整性的研究  

Study of comet assay for detecting sperm DNA integrity of patients with unexplained recurrent spontaneous abortion

作　　者：覃太洲 徐克惠[2] 刘莉 许文明[3] 李福平[2] QIN Taizhou;XU Kehui;LIU Li;XU Wenming;LI Fuping(Chengdu Women and Children＇s Central hospital, Chendu , Sichuan , 610031;West China Second University Hospital of Sichuan University;Joint Laboratory of Reproductive Medicine, Sichuan University-The Chinese U- niversity of Hong Kong Chengdu)

机构地区：[1]四川省成都市妇女儿童中心医院,610031 [2]四川大学华西第二医院 [3]四川大学-香港中文大学生殖医学联合实验室

出　　处：《中国计划生育学杂志》2018年第4期260-265,269,共7页Chinese Journal of Family Planning

基　　金：国家973计划项目(2014CB943301)

摘　　要：目的:采用单细胞凝胶电泳技术(又称彗星实验)对精子DNA完整性与不明原因复发性流产的相关性进行探讨,进一步明确精子DNA损伤的类型与不明原因复发性流产的关系。方法:收集2014年6月—12月在四川大学华西第二医院妇产科就诊的不明原因复发性流产女性的配偶作为流产组;同时收集有正常生育史的男性作为生育组。采用精液常规分析、中性彗星实验、碱性彗星实验等技术,检测精液常规参数和精子DNA完整性。同时分别人为诱导精子DNA单、双链损伤作为对照。结果:流产组和生育组各纳入50例男性,年龄差异无统计学意义,精液常规参数对比,流产组的精子浓度低于生育组[(91.0±56.8)×10~6/ml,(121.0±50.4)×10~6/ml](P<0.05),前向运动[(52.5±13.5)%,(54.3±10.8)%]、总活力[(58.9±12.7)%,(61.3±10.5)%]、正常形态精子百分率[(3.4±2.6)%,(3.3±2.2)%]两组差异均无统计学意义(P>0.05)。两组的精子DNA完整性对比,流产组比生育组拥有更高的精子DNA碎片指数[(29.5±4.3)%,(18.2±3.5)%]和精子单链DNA损伤率[(19.0±3.8)%,(8.9±3.0)%](P<0.05)。两组的精子双链DNA损伤率[(10.5±3.4)%,(9.2±2.4)%]未见差异(P>0.05)。结论:精子DNA损伤率过高,特别是DNA单链损伤过多可能是导致不明原因复发性流产的相关原因之一。精子常规参数可能对提示不明原因复发性流产的病因价值较小。Objective：To use single cell gel electrophoresis（or comet assay）to explore the connections between sperm DNA integrity and unexplained recurrent spontaneous abortion,and to further identify the relations between the type of sperm DNA breaks and unexplained recurrent spontaneous abortion.Methods：Fifty semen samples were obtained as abortion group,which were from spouses of patients with unexplained recurrent spontaneous abortion in the obstetrics and gynecology clinic of west China second hospital of Sichuan university from June to December 2014,and another fifty semen samples were collected as fertility group from male with healthy fertility.Three major methods were adopted to detect the semen parameters and sperm DNA integrity in this essay,which including semen analysis,neutral comet assay and alkaline comet assay.At the same time,results of which were compared with single-and double-stranded sperm DNA breaks induced by human factors.Results：There was no statistical significance different in age of spouses of patients.According to the comparison of semen parameters in the two groups,it is noticed that the sperm concentration in abortion group was much lower than that in fertility group（91.0 2± 56.8 vs.121.1±50.4）,which had statistically significant different（P 〈0.05）.There were no statistical significance（P 〉 0.05）in progressive motility（ 52.5 ± 13.5% vs.54.3±10.8%）,total motility（58.9±12.7% vs.61.3±10.5%）,and the percentage of morphology（normal forms）（3.4±2.6% vs.3.3±2.2%）.Compared with fertility group,abortion group had a significant higher sperm DNA fragmentation（SDF）index（29.5 ± 4.3 % vs.18.2± 3.5 %）and a higher rate of single-stranded sperm DNA breaks（19.0±3.8% vs.8.9±3.0%）.However,no statistical significance different（P〈0.05）was observed in the double-stranded sperm DNA breaks rates of both groups（10.5 ± 3.4 % vs.9.2 ± 2.4 %）.Conclusion：An excessively high rate of sperm DNA breaks,especially the rate of single-stranded sperm D

关 键 词：复发性流产 精子 DNA完整性 彗星实验 DNA单链损伤 

分 类 号：R714.21[医药卫生—妇产科学]