成年无精子症男性睾丸活检组织双向电泳分析技术的初步建立及其优化  

Preliminary Establishment and optimization of two-dimensional polyacrylamide gel electrophoresis for proteome analysis of the testicular tissue of the male patients with azoospermia

作　　者：李飞[1] 邹亚光[2] 赵亮[3] 周其赵[1] 李铁求[1] 郭文兵[1] 景晓玮[1] 李建明[3] 毛向明[1] 

机构地区：[1]南方医科大学南方医院泌尿外科,广州市510515 [2]南方医科大学南方医院口腔科,广州市510515 [3]南方医科大学南方医院病理科,广州市510515

出　　处：《实用医学杂志》2009年第24期4139-4142,共4页The Journal of Practical Medicine

基　　金：国家自然基金(编号:30772167);广州市科技计划项目(编号:2008J1-C131)

摘　　要：目的:建立并优化成年无精子症男性睾丸活检组织双向凝胶电泳图谱。方法:采用固相pH梯度等电聚焦为第一向,SDS-PAGE垂直电泳为第二向的双向电泳分离技术进行分离,凝胶经银染显色后,用Melanie 4软件分析双向凝胶电泳技术(2-DE)图谱。对聚焦参数的设置和样品的处理等步骤进行优化。结果:优化后成功地获得了成年无精子症男性睾丸活检组织分辨率高、重复性好的双向电泳图谱。结论:应用2-DE初步建立了成年无精子症男性睾丸活检组织分辨率较高且重复性好双向凝胶电泳图谱,为从睾丸蛋白质组学水平进一步研究无精子症奠定了基础。Objective To establish and optimize two-dimensional electrophoresis （2-DE） for proteome analysis of the testicutar tissue of the male patients with azoospemua. Methods Immobilized pH gradient （IPG） isoelectric focusing （IEF） electrophoresis on first dimension and SDS-PAGE on second dimension were used to separate the total proteins of the samples.Protein apois separated were dyed by silver staining.Protein profile were analyzed by Melanie 4 image analysis software, The sample preparation and the installation parameters of isoelectric focusing（lEF） were optimized. Results A high resolution and reproducible 2-DE image of the testieular tissue of the male patients with azoospermia is successfully obtained after optimizing. Conclusion Proteome profile of testicular tissues of azoospermia has been established, and it is the premise for further proteomie study of azoospermia

关 键 词：无精子症 双向凝胶电泳 睾丸 蛋白质组 

分 类 号：R698.2[医药卫生—泌尿科学] R532.21[医药卫生—外科学]