微小病变型肾病综合征激素耐药和敏感的比较尿蛋白质组学研究

目的 研究儿童微小病变型激素耐药与激素敏感型肾病综合征的尿中差异蛋白质表达,鉴定出肾病综合征激素耐药相关蛋白.方法 应用双向电泳相关技术结合基质辅助激光/电离飞行时间质谱仪技术,比较激素敏感和激素耐药微小病变型肾病综合征尿蛋白表达图谱寻找差异蛋白,对差异蛋白质斑点酶切,质谱分析,获得肽质量指纹图,通过蛋白质数据库进行蛋白检索分析.结果 成功地得到儿童微小病变型肾病综合征激素耐药尿蛋白双向电泳图谱.肾病综合征耐药型与激素敏感型蛋白表达比较,发现有30个蛋白质斑点显著差异改变.对14个差异蛋白质斑点酶切,质谱分析结合蛋白数据库检索获得12个蛋白,分别为驱动蛋白家族成员27、磷脂酰丝氨酸转移蛋白、大疱性类天疱疮抗原1异构体、α1蛋白酶抑制剂、Zn-α2糖蛋白、α1B糖蛋白、血清白蛋白前体、结合珠蛋白前体、类驱动蛋白样动力蛋白、白介素1受体相关激酶4、胞质动力蛋白、细胞角蛋白9.与激素敏感蛋白图谱比较,在激素耐药蛋白图谱上蛋白酶抑制剂、α1B糖蛋白、IRAK4表达下调,其余9种蛋白表达上调.结论 上述蛋白的鉴定将对激素耐药治疗靶位和耐药型肾病综合征诊断的分子标志物发现可能有所裨益.

Comparison of urinary proteomics between steroid-sensitive and steroid-resistant minimal change nephrotic syndrome in children

Objective To screen the differentially expressed proteins in the urine of children with steroid-sensitive and steroid-resistant minimal change nephrotic syndrome(SRINS and SSINS,respectively).Methods Urine samples were collected from 10 children with SRINS and 70 with SSINS as well as 30 healthy volunteers(control).Isoelectric focusing and two-dimensional electrophoresis in combination with matrix assisted laser desorption/ionization-time of flight(MALDI-TOF)mass spectrometry was performed for analysis of the urine proteins.Results and Conclusion In the urine samples,30 protein spots were identified to have differential expression between SRINS and SSINS.Further analysis of 14 protein spots identified 12 proteins expressing in SRINS,namely kinesin family member 27,PITPNB,bullous pemphigoid antigen,alpha-1 protease inhibitor,Zn-alpha-2GP,alpha-1B-glycoprotein,serum albumin precursor,haptoglobin precursor,kinesin like motor protein,IRAK4,cytoplasmic dynein and cytokeratin 9.Nine of these 12 proteins were up-regulated(U1-U3,U5,U7-U9,U11-U12)and 3 down-regulated(D4,D6,D10)in SRINS,suggesting that these proteins may serve as the potential therapeutic targets and as new diagnostic markers for steroid-resistant nephrotic syndrome.