罗丹明/碘化吡啶双染法检测精子线粒体膜功能的研究

目的:探讨罗丹明/碘化吡啶(Rh123/PI)双染法检测精子线粒体膜功能的可行性及临床意义。方法:收集63例精液标本,按WHO精液分析标准分成正常组(n=31)与异常组(n=32)。精子用Rh123/PI染色,流式细胞仪(FCM)分析。结果:正常组与异常组精子Rh123+PI-、Rh123-/PI+、Rh123-/PI-百分率均存在统计学差异(P均<0.05);Rh123+PI-与精子活动率呈正相关(r=0.549,P=0.000),与(a+b)级精子百分率呈正相关(r=0.531,P=0.000),与d级精子百分率呈负相关(r=-0.586,P=0.000);Rh123-/PI+与精子活率负相关(r=-0.586,P=0.000),与(a+b)级精子百分率负相关(r=-0.594,P=0.000),与d级百分率精子正相关(r=0.585,P=0.000),Rh123-/PI-仅与异常组的密度有相关性(r=-0.563,P=0.001)。结论:运用Rh123/PI双染法鉴定精子的线粒体膜功能具有可行性,可用于评估男性生育力。

Evaluation of Sperm Mitochondrial Function Using Rh123/PI dual Fluorescent Staining

Objective: To investigate the feasibility and clinical significance of detecting sperm mitochondrial function by using Rh123/PI dual fluorescent staining and flow cytometry analysis,and to explore the relationship between the results of Rh123/PI dual fluorescent staining and seminal parameters.Methods: Sixty-three semen samples were classified as normal(n=31) and abnormal(n=32) according to the World Health Organization guidelines.Rh123/PI dual fluorescent staining was then carried out to evaluate sperm mitochondrial function by flow cytometry analysis.Results: Significant differences in Rh123~( )PI~(-),Rh123~(-)/PI~( ) and Rh123~(-)/PI~(-) were detected between the normal and abnormal semen samples(P<(0.05)).There was a significant positive correlation between the Rh123~( )PI~(-) sperm and sperm motility and a significant inverse correlation between Rh123~( )PI~(-) and immotile sperm.But the Rh123~(-)PI~( ) sperm showed a contrary relationship with Rh123~( )PI~(-).A significant inverse correlation was also observed between the Rh123~(-)/PI~(-) sperm and sperm concentration in the abnormal group. Conclusion: Rh123/PI dual fluorescent staining and flow cytometry analysis can readily and quickly detect sperm mitochondrial function and be used to evaluate semen quality.Natl J Androl,2006,12(9):803-806