激素耐药型与激素敏感型肾病综合征儿童尿液双向电泳图谱分析  

Analysis of urine proteome profiles in children with nephrotic syndrome by two-dimensional polyacrylamide gel electrophoresis

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

机构地区：[1]中南大学湘雅二医院小儿肾脏病研究室,湖南长沙410011

出　　处：《中国实用儿科杂志》2007年第10期777-779,804,共4页Chinese Journal of Practical Pediatrics

基　　金：国家自然科学基金项目(编号30000184;30571983)

摘　　要：目的建立激素耐药型原发性肾病综合征(SRNS)儿童与激素敏感型原发性肾病综合征(SSNS)儿童尿液全蛋白双向凝胶电泳(2-DE)图谱,分析差异的蛋白质位点,为研究原发性肾病综合征(INS)糖皮质激素耐药的机制提供依据。方法病例来源于湘雅二医院儿科住院部,收集时间为2005-01-2005-10,入选的INS患儿依据中华医学会儿科学分会肾脏病学组制定的诊断标准,分为3组,每组5例。利用2-DE技术分离SRNS与SSNS(治疗前、后)儿童尿液总蛋白质,Image Master 2D V3.01分析软件进行凝胶图像分析。结果获得了图像清晰、分辨率高的SRNS组、SSNS(治疗前)与SSNS(治疗后)组儿童尿液2-DE图谱,3组点数分别为(206±8)个、(204±5)个、(55±5)个;SRNS组与SSNS组(治疗前)尿液2-DE凝胶共筛选出差异蛋白质点66个;SRNS组与SSNS组(治疗后)尿液2-DE凝胶共筛选出差异蛋白质点33个。结论该研究成功建立了分辨率高且重复性较好的SRNS与SSNS儿童尿液全蛋白的2-DE图谱;通过组间尿液2-DE图谱分析,发现了一些可能与INS激素耐药或敏感有关的蛋白质位点,为进一步运用蛋白质组学技术寻找与INS激素耐药抑或激素敏感相关的蛋白质打下了基础。Objective To investigate a proteome profile of urine in children with steroid-resistant or -sensitive idiopathic nephrotic syndrome （INS） by two-dimensional polyacrylamide gel electrophoresis （2-DE） ,in order to search for the different proteins for the further study of steroid-resistant mechanisms in INS. Methods Collect urine samples from children with steroid- resistant INS （SSNS） ,steroid-sensitive INS （SRNS） before and after steroid therapy,then use 2-DE and Image Master 2D V3.01 software to analyze the different expression of proteins in 2-DE gels. Results Mean protein spots of the 2-DE gels were （ 206 ± 8 ）, （ 204 ± 5 ）, （55 ± 5 ） in children with SRNS, SSNS before or after steroid treatment. Sixty-six different protein spots were screened between the SRNS and SSNS group before steroid therapy. Thirty-three different protein spots were screened between SRNS and SSNS after steroid therapy. Conclusion A distinct and reproducible 2-DE profile of urine proteome is successfully established in children with steroid-resistant and steroid-sensitive INS. By the differentiated analysis ,the differences of urine proteome profiles are discovered between steroid- resistant and steroid-sensitive INS,and some of these special proteins may be related to the mechanism of steroid-resistant and steroid-sensitive in INS.

关 键 词：原发性肾病综合征 糖皮质激素 尿液 双向凝胶电泳 儿童 

分 类 号：R72[医药卫生—儿科]