肾移植术后尿蛋白二维电泳图谱的建立  

Establishment of two dimensional electrophoregram of urine proteome in patients with acute rejection after renal transplantation

作　　者：贾雄飞[1,2] 贺伟峰[1] 毛小琴[3] 甘成军[1] 罗高兴[1] 黄正根[1] 袁顺宗[1] 王晓娟[1] 彭旭[1] 程文广[1] 谭江琳[1] 吴军[1] 

机构地区：[1]第三军医大学西南医院重庆市疾病蛋白质组重点实验室 [2]解放军昆明总医院检验科,昆明650032 [3]重庆医科大学检验系

出　　处：《中国实验诊断学》2008年第5期613-615,共3页Chinese Journal of Laboratory Diagnosis

基　　金：国家自然科学基金资助项目(30500447)

摘　　要：目的近年来诊断急性排斥反应的发生,一直朝着寻找非侵入性的检测方向努力,尿液中的蛋白可以为肾脏疾病病理变化提供线索。本次实验的目的旨在建立肾移植术后病人尿蛋白二维电泳图谱。方法利用二维凝胶电泳(Two-dimensional Gel Electrophoresis,2-DE)技术,以固相pH梯度等电聚焦为第一向,改良等电聚焦的电压在3500V,总伏小时为14000 Vh;12%的SDS-PAGE进行第二向电泳,银染显色。结果得到了令人满意的肾移植术后发生急性排斥反应患者尿蛋白二维电泳图。结论此方法,可为后续利用蛋白质组技术,研究急性排斥反应诊断标志物提供可靠的技术支持。Objective Aim In recent years,scientists are looking for noninvasive methods to diagnose the occurrence of acute re- jection, and urine proteins can reflect the pathological change of kidney diseases. Our experiment aims to establish two-dimensional electrophoregram of urine proteins of patients after renal transplantation. Methods Two-dimensional gel eleetrophomsis was carried out to obtain electrophomgram.Briefly,modified IEF was carried out with immobilized pH gradients. The IPG gels were focused at 3 500 V and total VH was constantly 14 000 Via. The gels were then submitted to SDS-PAGE and giver stained. Results Satisfying two-dimensional electrophoregram of urine proteins of patients with acute rejection after renal transplantation was obtained. Conclusion Using 2-DE technique to establish two-dimensional electrophomgram, this provides the theoretical basis and technical support for looking for diagnosis marker of acute rejection by proteomic approach.

关 键 词：肾移植 急性排斥反应 蛋白质组 二维电泳 

分 类 号：R699[医药卫生—泌尿科学]