【摘要】 目的探讨常规精液冷冻技术对人类精子线粒体DNA的影响。方法收集符合精子库捐精条件的正式志愿者精液样品,将每份样品分为2份:1份进行精液冷冻复苏处理,为实验组;1份新鲜精液,为对照组。采用Markler计数板联合CASA法评估冷冻复苏前后精子活力;两组精液均采用实时荧光定量PCR和长链PCR技术,分别检测精子线粒体DNA的拷贝数和完整性。结果共收集22份精液标本,纳入志愿者年龄为(27.8±3.0)岁,禁欲天数(6.1±0.9)d;精液体积(5.0±1.5)ml,精子浓度(75.8±15.8)×106/ml,前向运动精子百分比为(68±6)%,总活动精子复苏率为(72±8)%。冷冻保存后,精子活力显著降低:前向运动精子百分比减少[(49.0±6.5)%vs.(68.0±6.1)%,P<0.05),平均路径速率(VAP)[(35.8±6.8)vs.(46.8±9.5),P<0.05]、直线速率(VSL)[(27.3±3.3)vs.(35.1±8.3),P<0.05]和曲线速率(VCL)[(57.6±6.9)vs.(91.8±10.2),P<0.05]较前下降。与新鲜精液相比,冷冻复苏后精子的线粒体DNA拷贝数显著增加[(10.12±8.41)vs.(5.66±5.53),P<0.05],完整性比较无统计学差异[(29.69±15.04)vs.(32.78±16.0),P=0.077]。结论在正式捐精志愿者人群内,常规精液冷冻技术降低精子活力,增加人类精子mtDNA的拷贝数,但未显著改变mtDNA的完整性。更多还原

【Abstract】 Objective:To detect the effect of routine semen cryopreservation on human sperm mitochondrial DNA.Methods:The semen samples from normal donors were collected in Beijing Human Sperm Bank.Each sample was divided into 2 parts:the cryopreservation experience group and the control group(fresh semen without cryopreservation).The sperm motility was examined by Marker and CASC.The copy number and integrity of sperm mtDNA were detected by real-time fluorescence quantitative(Q-PCR)and long polymerase chain reaction(L-PCR)respectively.Results:A total of 22 sperm samples were collected.The age of the donors was(27.8±3.0)years,thenumber of abstinence days(6.1±0.9)days,the volume of semen(5.0±1.5)ml,the sperm concentration(75.8±15.8)×106/ml,the percentage of the progressive motility sperm(68±6)%,and the recovery rate of motile sperm(72±8)%.After cryopreservation,sperm motility including the percentage of RP[(49.0±6.5)% vs.(68.0±6.1)%,P<0.05]and sperm motility parameters VAP[(35.8±6.8)vs.(46.8±9.5),P<0.05],VSL[(27.3±3.3)vs.(35.1±8.3),P<0.05]and VCL[(57.6±6.9)vs.(91.8±10.2),P<0.05]was decreased significantly.The copy number of mtDNA of sperm with cryopreservation was increased significantly[(10.12±8.41)vs.(5.66±5.53),P<0.05].However,the sperm mtDNA integrity was not changed after cryopreservation[(29.69±15.04)vs.(32.78±16.0),P=0.077].Conclusions:Sperm cryopreservation technology decreases the sperm motility and increases the copy number of human sperm mtDNA,but does not change the sperm mtDNA integrity,in the eligible sperm donors.更多还原