双向电泳分析精索静脉曲张不育患者精子蛋白质的表达差异

Differential expression analysis of sperm proteins in infertile men with varicocele using two-dimensional gel electrophoresis

目的：比较分析正常生育男性与精索静脉曲张（varicocele, VC）不育患者差异表达的精子蛋白质。方法应用计算机辅助精子分析系统行精液分析，采取Percoll密度梯度离心法提取正常生育男性与VC不育患者精子，利用双向电泳（two-dimensional electrophoresis,2-DE）分离精子蛋白质，建立并优化精子蛋白质2-DE图谱，利用PDQuest8.0软件分析两组蛋白质的差异表达。结果正常生育组精子活力高于VC不育组，两组比较差异有统计学意义（P＜0.05），两组精子密度比较差异无统计学意义（P＞0.05）。获得了分辨率和重复性均较好的2-DE图谱，两组差异点共36个，VC不育组有11个蛋白位点表达上调，19个蛋白位点表达下调，2个在VC不育组特异性表达，4个在正常生育组特异性表达。结论 VC不育患者与正常生育男性的精子蛋白存在差异，这些差异蛋白质的分离为深入探讨VC导致不育的分子机制奠定了基础。

Objective To study the differential expression of sperm proteins between fertile men and infertile men with varicocele（VC）. Methods Sperm samples were analyzed by computer-assisted semen analysis system. Sperm proteins were extracted by percoll density gradient centrifugation, and the separation of total proteins was performed by two-dimensional electrophoresis（2-DE）. The differentially expressed proteins of two groups were analyzed with PDQuest software. Results The sperm motility in infertile men with VC demonstrated a significant decrease compared with the fertile men（P〈0.05）, and there was no significant difference between two groups in sperm concentration（P〉0.05）. Protein profile of sperm was acquired with clear background, high resolution and better reproduction.Total of 36 different proteins were found, including 11 upregulated protein and 19 downregulated proteins in infertile men with VC group, and two proteins only expressed in infertile men with VC group, the other four proteins only expressed in fertile men group. Conclusion There are differential expression of sperm proteins between fertile men and infertile men with VC. Differential proteins analysis will be helpful to explore the molecular mechanism of VC-associated male infertility.