流式细胞术在精子功能检测中的应用  

Application of Flow Cytometry to the Evaluation of Semen Quality

作　　者：华咏[1] 钱宪明[1] 陈炳官[1] 杨建华[1] 吴晓云 马亮[1] 陈翠玲[1] 

机构地区：[1]同济大学附属东方医院中心实验室,上海200120 [2]上海市博康生殖医院精液化验室,上海200032

出　　处：《中华男科学杂志》2006年第7期608-611,共4页National Journal of Andrology

摘　　要：目的:研究流式细胞术在精子功能评价中的作用。方法:对104例男性不育患者和10例正常生育男性的精液标本进行精液量、精子密度、活动率、畸形率等精液常规分析(CASA法),对精子存活率、染色质结构和线粒体膜电位等进行流式细胞术分析,并将检测结果通过SAS软件进行统计学分析。结果:成组U检验结果表明:不育患者的精子与正常生育男性精子相比除了具有密度低(U=2.51,P=0.014 3)、活动率差(U=3.44,P=0.001)、畸形率高(U=-5.88,P<0.000 1)等特点外,经流式细胞术测定精子结果表明不育男性的精子质膜完整率(U=4.72,P<0.000 1)、线粒体膜电位正常率(U=-2.53,P=0.030 9)和染色质完整率(αt:U=-3.82,P=0.000 3;SDαt:U=-3.98,P=0.000 1;COMPαt:U=-3.57,P=0.000 5)均显著低于正常生育男性。多元逐步回归分析结果表明:精子质膜完整性与精子活动率(t=1.66,P=0.101 6)、精子的线粒体膜电位与精子活动率(t=3.33,P=0.001 4)、精子染色质的完整性与精子活动率(t=3.24,P=0.001 9)均呈正相关,而精子的线粒体膜电位与精子颈、尾部的畸形率呈负相关(t=-3.44,P=0.001)。结论:流式细胞仪测定对精子质量评定意义显著,对男性不育的诊断和治疗也具有较高的参考价值。Objective： To evaluate the role of flow cytometry in semen assessment. Methods： Semen samples from 104 infertile male patients （as the case group） and 10 fertilized donors （as the control group） were analyzed for the volume of ejaculate and sperm concentration, motility and atypical morphology by computer-assisted semen analysis （ CASA）, the viability, chromatin structure and mitochondrial membrane potential（MMP） of the sperm stained by SYBR-14/PI, AO and JC-1 respectively, and assessed with flow cytometry. The results were analyzed through SAS software. Results ： U tests indicated that the semen from the infertile patients had not only lower concentration（ U = 2.51, P = 0.0143 ）, lower motility （ U = 3.44, P = 0. 001 ） and higher rate of atypical morphology （ U = -5.88, P 〈0.000 1）, but also lower viability（ U =4.72, P 〈0.000 1）, MMP（U= -2.53, P =0.030 9）, and chromafin integrity （ αt ： U = - 3.82, P = 0.000 3 ; SDαt： U = - 3.98, P = 0.000 1 ; COMPort： U = - 3.57, P = 0.000 5 ）. The multiple stepwise regression analysis revealed that sperm motility was positively correlated with sperm membrane integrity（ t = 1.66, P = 0. 1016 ）, sperm MMP（ t = 3.33, P =0.001 4） and sperm acrosome integrity （t = 3.24, P = 0.001 9 ）, while sperm MMP was negatively correlated with the rates of sperm neck and tail defects （ t = - 3.44, P = 0.001 ）. Conclusion ： Flow cytometry plays a significant role in the evaluation of the quality of human sperm, and can be adopted as a useful tool in the diagnosis and treatment of male infertility.

关 键 词：精子 流式细胞术 质膜完整性 染色质结构分析 线粒体膜电位 

分 类 号：R698.2[医药卫生—泌尿科学]