儿童激素耐药型与激素敏感型肾病综合征尿蛋白组学的初步研究  

Preliminary analysis of urinary proteomics in children with steroid-resistant and steroid-sensitive nephrotic syndrome

作　　者：黄爱文[1] 何庆南[1] 周频[1] 丁娟娟[1] 易著文[1] 

机构地区：[1]中南大学湘雅二医院儿科、小儿肾脏病研究室,湖南长沙410011

出　　处：《中国当代儿科杂志》2009年第5期341-345,共5页Chinese Journal of Contemporary Pediatrics

基　　金：国家自然科学基金项目(编号30000184,30571983)

摘　　要：目的筛选和鉴定激素耐药型(SRNS)与激素敏感型原发性肾病综合征(SSNS)尿液中差异表达的蛋白质,寻找可能与肾病综合征激素耐药和激素敏感相关的蛋白质。方法SSNS治疗前后病例、SRNS、正常对照各5例,提取尿液全蛋白,采用双向凝胶电泳建立全蛋白凝胶图谱;银染后用Image Master 2D V3.01软件分析差异表达蛋白质点,利用MALDI-TOF-MS分析获得肽质量指纹图谱,通过Mascot软件进入NCBI数据库,鉴定出差异表达蛋白质。结果SRNS组与SSNS治疗前组尿液2-DE凝胶比较有差异蛋白质点66个,其中仅出现于SRNS组24个、SSNS治疗前组27个;SSNS治疗后组与正常对照组尿液2-DE凝胶比较有差异蛋白质点75个,其中仅出现于SSNS治疗后组11个。对18个差异蛋白质点进行分析,鉴定出α1-抗胰蛋白酶、转铁蛋白、带异类电荷糖蛋白复合体等9类差异表达的蛋白质。结论成功鉴定了SRNS和SSNS的尿液中存在9类差异表达的蛋白质,部分蛋白可能是SRNS或SSNS的标志物。Objective To study and identify the protein markers in the urine of children with steroid-sensitive （SSNS） and steroid-resistant nephrotic syndrome （SRNS）. Methods Total urinary proteins were extracted from children with SSNS before and after steroid therapy, SRNS, and healthy children （ n ： 5 in each group）. Urinary proteins were separated by immobilized pH gradient based on two-dimensional gel electrophoresis （2-DE）. The silver-stained 2-DE gels were scanned with digital Image Scanner and analyzed with Image Master 2-DE Elite 3.01 software. Peptide mass fingerprint （PMF） of differential protein spots was obtained with MALDI-TOF-MS. Proteins were identified by Mascot software based on NCBI protein database. Results There were 66 spots with different expression of protein between SRNS children and SSNS children before steroid therapy, and 24 spots and 27 spots only occurred in SRNS children and SSNS children before steroid therapy, respectively. There were 75 spots with different expression of protein between SSNS children after steroid therapy and healthy controls, and 11 spots only occurred in SSNS children after steroid therapy. Eighteen protein spots with different expression （6 spots in each nephrotic group） were chose and analyzed by MALDI- TOF-MS, and 9 types of proteins were identified. Conclusions Nine types of urinary proteins with different expression （6 spots in each nephrotic group） were identified between SRNS and SSNS children, and they might be the biomarkers for SRNS or SSNS.

关 键 词：原发性肾病综合征 激素耐药 蛋白组学 儿童 

分 类 号：R726.9[医药卫生—儿科]