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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

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【作者】 丁娟娟何庆南周频张磊易著文

【Author】 DING Juan-Juan,HE Qing-Nan,ZHOU Pin,et al.Department of Pediatrics,the Second Xiangya Hospital,Central South University,Changsha 410011,China

【机构】 中南大学湘雅二医院小儿肾脏病研究室中南大学湘雅二医院小儿肾脏病研究室 湖南长沙410011湖南长沙410011

【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。

【Abstract】 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.

【基金】 国家自然科学基金项目(编号30000184,30571983)
  • 【文献出处】 中国实用儿科杂志 ,Chinese Journal of Practical Pediatrics , 编辑部邮箱 ,2007年10期
  • 【分类号】R726.9
  • 【被引频次】1
  • 【下载频次】189
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