SYBR-14/PI双染法流式细胞术检测精子质膜完整性的研究  

Evaluation of sperm plasma membrane integrity by SYBR-14/PI dual fluorescent staining and flow cytometry

作　　者：周鑫[1,2] 夏欣一[2] 曹奇[2] 康宁[1,2] 范晓博[1,2] 邵永[2] 李瑛[3] 黄宇烽[1,2] 

机构地区：[1]南京师范大学生命科学学院,江苏南京210046 [2]南京军区南京总医院解放军临床检验医学研究所,江苏南京210002 [3]江苏省计划生育科学技术研究所,江苏南京210036

出　　处：《中华男科学杂志》2010年第7期589-593,共5页National Journal of Andrology

基　　金：江苏省科技厅生殖健康研究技术服务平台项目(BM2008151)~~

摘　　要：目的:探讨应用荧光染料SYBR-14/碘化丙啶(SYBR-14/PI)双色标记法进行流式细胞术检测精子质膜完整性的可行性及其临床意义。方法:收集208例男性精液标本,按WHO精液分析标准分为正常组(n=31)与异常组(n=177)。通过计算机辅助精液分析系统进行精液常规分析。精液标本洗涤处理后经SYBR-14/PI双染后上流式细胞仪(FCM)分析,用发绿色荧光精子百分率(SYBR-14+/PI-%)表示质膜完整精子(PMI)的比例。结果:正常组与异常组SYBR-14+/PI-与SYBR-14-/PI+精子百分率均存在统计学差异(P均<0.05)。正常组精子SYBR-14+/PI-%为(55.66±20.64)%,显著高于异常组[(39.71±19.21)%,P=0.000]。208例标本中,SYBR-14+/PI-%与精子活动率呈显著正相关(r=0.408,P=0.000),与(a+b)级精子百分率呈显著正相关(r=0.398,P=0.000),与d级精子百分率呈显著负相关(r=-0.413,P=0.000);SYBR-14-/PI+%与精子活动率呈显著负相关(r=-0.380,P=0.000),与(a+b)级精子百分率呈显著负相关(r=-0.397,P=0.000),与d级精子百分率呈显著正相关(r=0.385,P=0.000);SYBR-14+/PI+%与精子活动率呈正相关(r=0.172,P=0.013),与(a+b)级精子百分率呈正相关(r=0.177,P=0.011),与d级精子百分率呈负相关(r=-0.164,P=0.018)。结论:应用流式细胞术SYBR-14/PI双染法检测精子质膜完整性具有可行性,可用于评估男性生育力。Objective： To investigate the feasibility and clinical significance of detecting the plasma membrane integrity （PMI） of sperm by SYBR-14/PI fluorescent staining and flow cytometry. Methods ： A total of 208 semen samples were divided into a normal （n = 31 ） and an abnormal group （n = 177 ）, subjected to conventional computer-assisted semen analysis （CASA）, and then evaluated for sperm PMI by flow cytometry after washed and SYBR-14/PI dual fluorescent staining. The percentage of sperm with PMI was indicated as that of the sperm emitting green fluorescence （ SYBR-14 ^＋/PI - % ）. Results ： Significant differences were detected in SYBR-14 ^＋/PI- and SYBR-14^-/PI ＋ between the normal and abnormal groups （P 〈 0.05 ）, with the SYBR-14 ^＋/PI-% signitleantly higher in the former （[55.66 ± 20. 641%） than in the latter （[39.71 ± 19.211%） （P = 0.000）. The SYBR-14 ^＋/PI-% showed significant positive correlations with sperm motility （ r = 0. 408, P = 0. 000） and the percentage of grade a ＋ b sperm （r = 0.398, P = 0.000）, and a significant negative correlation with the percentage of grade d sperm （r = -0.413, P = 0.000） ; the SYBR-14^ -,/PI^＋ % exhibited significant negative correlations with sperm motility （r = - 0. 380, P = 0. 000） and the percentage of grade a ＋ b sperm （ r = - 0.397, P = 0. 000）, and a significant positive correlation with the percentage of grade d sperm （ r = 0.385, P = 0. 000） ; the SYBR-14 ^＋/PI ＋ % showed positive correlations with sperm motility （r = 0. 172, P = 0. 013） and the percentage of grade a ＋ b sperm （r = 0. 177, P = 0. 011）, and a negative correlation with grade d sperm （r = -0. 164, P = 0. 018）. Conclusion： SYBR-14/PI dual fluorescent staining and flow cytometry could be readily used to detect sperm PMI and evaluate male reproductivity. Natl J Androl, 2010, 16（7） ： 589 -593

关 键 词：SYBR-14/PI双染法 精子 质膜完整性 流式细胞术 

分 类 号：R698.2[医药卫生—泌尿科学]