儿童原发性肾病综合征激素敏感与耐药者尿蛋白质组学比较  

A comparison of urine proteomics between children with steroid-sensitive and steroid-resistant nephrotic syndrome

作　　者：王秋霞[1] 张婕[1] 相虹[1] 张璐洁[1] 卢红艳[1] 

机构地区：[1]江苏大学附属医院儿科,江苏镇江212001

出　　处：《现代医学》2016年第1期27-32,共6页Modern Medical Journal

基　　金：江苏省科技创新与成果转化(生命健康科技)专项资金项目(BL2012058)

摘　　要：目的：比较原发性肾病综合征中激素敏感型及激素耐药型儿童的尿蛋白质谱,寻找可能的激素敏感和激素耐药的尿蛋白标志物。方法：取原发性肾病综合征儿童中激素敏感型4例和激素耐药型4例的尿液各100~200 ml,分别处理尿液蛋白质后取等量的尿蛋白,分别混合成激素敏感组和激素耐药组尿蛋白,进行双向电泳。用基质辅助激光解吸/电离飞行时间质谱（MALDI-TOF-MS）方法对差异明显的蛋白点进一步鉴定,得到蛋白质点的肽质量指纹图谱（PMF）,通过Mascot软件进入NCBI数据库,鉴定出差异表达的蛋白质。结果：原发性肾病综合征激素敏感组和激素耐药组凝胶的平均蛋白点数分别为37±3和40±4;两组间尿蛋白质谱有差异,差异蛋白点有13个;对其中差异明显的10个蛋白点进一步鉴定得到9个蛋白,激素敏感组鉴定出4个蛋白点,激素耐药组鉴定出5个蛋白点。结论：通过双向电泳、MALDI-TOF-MS分析,筛选出激素敏感型和激素耐药型肾病综合征儿童尿液中存在的9种差异表达的蛋白。Objective： Two-dimensional gel electrophoresis（ 2-DE） gels of urine protein were compared between children with steroid-sensitive nephrotic syndrome（ SSNS） and with steroid-resistant nephrotic syndrome（ SRNS） to find out the differential protein spots. Then the differential protein spots were identified in order to find out the probable urine biomarkers of children with SSNS and with SRNS. Methods： Urine specimens were collected from 4children with SSNS and 4 children with SRNS separately. Equal proteins from every child were mixed according to the SSNS group and the SRNS group. The proteins were separated by 2-DE. The 2-DE gels stained by Coomassie brilliant blue were firstly scanned by digital Image Scanner. Then Image Master 2D Platinum software was used to analysis the scanned images. Matrix assisted laser desorption / ionization time-of-flight mass spectrometry（ MALDITOF-MS） was used to obtain the peptide mass fingerprints of differential protein spots. The results were searched in NCBI protein database by Mascot software. Results： Mean protein spots of SSNS group and SRNS group in the 2-DE gels were 37 ± 3 and 40 ± 4,respectively. Thirteen protein spots expressed differentially between SSNS and SRNS were identified. Among the 13 protein spots,10 spots were chosen and analyzed by MALDI-TOF-MS and 9proteins were identified. Conclusion： There were 9 differential proteins expressed in SSNS group and SRNS group,which might relate to the mechanism of steroid-resistance.

关 键 词：肾病综合征 尿液 激素敏感 激素耐药 基质辅助激光解吸/电离飞行时间质谱 

分 类 号：R692[医药卫生—泌尿科学]