尿蛋白组学在儿童微小病变型肾病综合征中的初步研究  

Preliminary analysis on urinary proteomics of children with minimal change nephrotic syndrome

作　　者：黄艳军[1] 黄松明[1] 张爱华[1] 费莉[2] 徐颂周[2] 陈荣华[2] 

机构地区：[1]南京医科大学附属南京儿童医院肾脏科,江苏南京210008 [2]南京医科大学小儿肾脏病研究中心,江苏南京210029

出　　处：《中国现代医学杂志》2006年第20期3152-3155,共4页China Journal of Modern Medicine

基　　金：江苏省自然科学基金资助(BK2002030)

摘　　要：目的研究儿童微小病变型肾病综合征尿蛋白组分析所需的固相pH梯度等电聚焦双向电泳及相关技术。方法冷丙酮萃取肾病综合征患儿尿蛋白,去除白蛋白等高丰度蛋白,以固相IPGpH梯度等电聚焦为第一向,SDS均一胶(10.00%)的垂直电泳为第二向,银染后PDQuest软件进行分析,质谱分析。结果成功地得到儿童微小病变型肾病综合征尿蛋白双向电泳图谱。PDQuest7.3软件进行图像分析,检测到400个蛋白点。正常尿蛋白检测到112个蛋白点。肾病综合征与正常尿蛋白图谱相比差异蛋白表达明显的有20个,质谱鉴定出5个蛋白,分别是Alpha-1-antitrypsin、PITPNB、Zn-alpha-2-glycoprotein、Haptoglobin和Alpha-1B-gly-coprotein。结论用尿蛋白组学对儿童微小病变型肾病综合征的尿液进行研究是可行的。[Objective ] The aim of the research is to establish the two-dimensional gel electrophoresis and PDQUEST software based image analysis for the urinary proteome of children with minimal change nephrotic syndrome. [Methods] The urinary protein of children nephrotic syndrome was isolated by cold acetone precipitation, treated to remove high abundance proteins by Aurum Serum Protein Kit .The rehydrated IPG strips containing protein samples were isoelectric focusing （IEF）, after IEF, the equilibrated strips are transferred to the vertical SDS gel. The silver stained gels were scanned by UMAX PowerLook 1120 densitometer, following PDQuest 7.3 analysis, the differentially expressed protein spots were found and excised from gel and digested with trypsin under optimal conditions. Masses of tryptic-digested peptides were acquired with a Voyager DA-STR matrix assisted laser desorption ionization time of flight （MALDI-TOF） mass spectrometer. [Results] Using the proper method stated above, satisfactory 2DGE maps of urinary protein was obtained and the preliminary analysis results were reported. A total of 400 protein spots were visualized in the patients with minimal change nephrotic syndrome, about 100 protein spots in the normal urine. Nine spots of proteins in gel were identified as Alpha-1-antitrypsin precursor, Zn-alpha-2-glycoprotein, Alpha-1B-glycoprotein, Haptoglobin, and PITPNB protein. [Conclusion] Urinary proteomics applied to the study of urine protein of children minimal change nephrotic syndrome is feasible.

关 键 词：尿蛋白质组 双向电泳 图像分析 质谱 

分 类 号：R692[医药卫生—泌尿科学]