原发性肾病综合征儿童尿液双向电泳图谱的建立

Establishment of urinary proteomic map on two-dimensional polyacrylamide gel electrophoresis in children with idiopathic nephrotic syndrome

目的建立与优化原发性肾病综合征(INS)儿童尿液全蛋白双向凝胶电泳技术(2-DE)。方法采用4种不同的方法制备蛋白质样本,以固相pH梯度等电聚焦为第一向,12%的SDS-PAGE进行第二向电泳,银染显色。结果以丙酮浓缩蛋白后透析提纯法,制备蛋白质标本,300μg为上样量时,可获得图像清晰、分辨率高、重复性好的儿童INS尿液2-DE图谱。结论建立了稳定的原发性肾病综合征儿童尿液双向凝胶电泳技术,为进一步开展疾病的尿液蛋白质组学研究奠定了基础。

Objective To establish a urinary proteomic map on two-dimensional polyacrylamide gelelectrophoresis（2- DE） in children with idiopathic nephrotic syndrome （INS）. Methods The proteins from INS children were purified by four various means, separated by 2-DE, and stained by silver. Results The sequential preparation of urinary proteins by acetone precipitation and dislysis, when the sample was 300 μg, resulted in a clear background, well-resolved and reproducible 2-DE urinary protemic map in children with INS. Conclusions A steady 2-DE technique for urinary protemic map in children with INS was established, which can be effectively applied in urinary proteomics of the disease.