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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 establish a 2-DE map of children with steroid-resistant primary nephrotic syndrome (SRNS) and children with steroid-sensitive nephrotic syndrome (SSNS) and analyze the difference of protein level Point, to provide a basis for studying the mechanism of glucocorticoid resistance in primary nephrotic syndrome (INS). Methods The cases were from Department of Pediatrics, Second Xiangya Hospital, collected from January 2005 to June 2005. The selected children with INS were divided into three groups according to the diagnostic criteria developed by the Nephrology Group of Pediatric Branch of Chinese Medical Association. Group 5 cases. The 2-DE technique was used to separate urine total protein from SRNS and SSNS (before and after treatment). Image Master 2D V3.01 software was used for gel image analysis. Results The 2-DE maps of urine of SRNS, SSNS (pre-treatment) and SSNS (post-treatment) children with clear images and high resolution were obtained. The scores of the 2 groups were 206 ± 8 and 204 ± 5, (55 ± 5). There were 66 differential protein spots in urinary 2-DE gels of SRNS group and SSNS group (before treatment). Urinary 2-DE gels of SRNS group and SSNS group A total of 33 differentially expressed protein spots were screened. Conclusions The study successfully established a 2-DE map of urinary total protein in SRNS and SSNS children with high resolution and good reproducibility. Based on the 2-DE analysis of urine among groups, some 2-DE patterns that may be resistant to INS hormone or Sensitive related protein sites lay the foundation for further application of proteomics techniques in finding proteins related to INS hormone resistance or hormone sensitivity.