男性不育患者精子染色体畸变及精子DNA完整性分析

目的 探讨男性不育患者精子染色体和精子DNA完整性的改变.方法 精子染色质扩散(sperm chromatin dispersion,SCD)实验分析精子DNA碎片,正常生育男性(对照组)32名,特发性严重少弱精子症患者(idiopathic severe oligoasthenozoospermia,ISOA)19例,妻子不明原因反复自然流产(unexpbined recurrent miscarriage,URM)38例;多色荧光原位杂交(fluorescent in situ hybridization,FISH)技术检测URM妇女丈夫(n=12)、ISOA不育者(n=10)及对照组(n=5)精子13、21、18、X和Y染色体畸变.结果 对照组、ISOA不育者及URM妇女丈夫精子13、18和21染色体数目总体异常的比率分别为1.29%、4.02%和3.91%,而X和Y染色体数目总体异常的比率分别为0.61%、2.03%和1.98%,与对照组比较差异均有统计学意义(P＜0.01).SCD实验分析精子DNA碎片,ISOA不育患者(n=19)及URM妇女丈夫(n=38)精子DNA碎片比例平均为40.7%±17.8%和22.1%±10.3%,均显著性高于对照组(12.1%±5.2%,P＜0.01).FISH精子染色体(13、21、18、X和Y探针)数目畸变率与精子DNA碎片比率呈正相关(r=0.874,P＜0.01,n=27),精子DNA碎片比率与精子密度及前向运动精子率呈负相关(r=-0.571,P＜0.01,和r=-0.616,P＜0.01,n=89),与畸形精子比率呈正相关(r=0.637,P＜0.01,n=89).结论 精子染色体畸变率和精子DNA碎片比率增高,可能是ISOA和妻子URM不育男性的原因之一,精子DNA损伤筛查町能为特发性男性不育患者提供有用的信息.

Analysis of sperm chromosomal abnormalities and sperm DNA fragmentation in infertile males

Objective To investigate changes in sperm chromosome and sperm DNA integrity of infertile males.Methods The level of DNA fragmentation was determined by Sperm Chromatin Dispersion (SCD) test in infertile males with idiopathic severe oligoasthenozoespermia (ISOA,n=19),couples with unxplained recurrent miscarriage (URM,n=38) and adult healthy fertile men (control group,n=32).Multi-color fluorescence in situ hybridization (FISH)was performed with probes specific for chromosomes 13,18,21,X and Y in the control group (n=5),the ISOA (n=10) andtheURM(n=12).Results Patients with LSOA and URM showed a signiiicantly higher abnormality with total rate of 4.02% (n=19) and 3.91% (n=38) for chromosomes 13,18 and21,and2.03%,1.98% for chromosomes X and Y,respectively,in their spermatozoa compared to control (1.29% and 0.61%,P＜0.01).A significantly higher proportion of total sperm DNA fragmentation was detected in patients with ISOA (40.7%± 17.8%) and URM (22.1% ±10.3 %) of sperm compared to the control group (12.1%± 5.2%,P＜0.01).Moreover,a positive correlation was found between the rate of sperm chromosomal aberration and the rate of sperm DNA fragmentation (r=0.874,P＜0.01,n=27).There were significant correlation between sperm DNA fragmentation and sperm density,sperm motility and abnormal sperm (r=-0.571,r=-0.616 and r=0.637,respectively,P＜0.01).Conclusion result indicates that spermatozoa from patients with ISOA and URM contain greater DNA fragmentation and chromosomal aneuploidy and may lead to male infertility.Screening for sperm DNA damage may provide useful information in the diagnosis of male idiopathic infertility.