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目的:比较原发性肾病综合征中激素敏感型及激素耐药型儿童的尿蛋白质谱,寻找可能的激素敏感和激素耐药的尿蛋白标志物。方法:取原发性肾病综合征儿童中激素敏感型4例和激素耐药型4例的尿液各100~200 ml,分别处理尿液蛋白质后取等量的尿蛋白,分别混合成激素敏感组和激素耐药组尿蛋白,进行双向电泳。用基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF-MS)方法对差异明显的蛋白点进一步鉴定,得到蛋白质点的肽质量指纹图谱(PMF),通过Mascot软件进入NCBI数据库,鉴定出差异表达的蛋白质。结果:原发性肾病综合征激素敏感组和激素耐药组凝胶的平均蛋白点数分别为37±3和40±4;两组间尿蛋白质谱有差异,差异蛋白点有13个;对其中差异明显的10个蛋白点进一步鉴定得到9个蛋白,激素敏感组鉴定出4个蛋白点,激素耐药组鉴定出5个蛋白点。结论:通过双向电泳、MALDI-TOF-MS分析,筛选出激素敏感型和激素耐药型肾病综合征儿童尿液中存在的9种差异表达的蛋白。
OBJECTIVE: To compare urinary protein profiles of hormone-sensitive and steroid-resistant children with primary nephrotic syndrome and to search for possible hormone-sensitive and hormone-resistant urinary protein markers. Methods: The children with primary nephrotic syndrome hormone-sensitive 4 cases and hormone-resistant 4 cases of urine 100 ~ 200 ml, respectively, after treatment of urine protein and take the same amount of urine protein were mixed into a hormone-sensitive Group and hormone resistance group urine protein, two-dimensional electrophoresis. Differentially identified protein spots were further identified by matrix-assisted laser desorption / ionization time-of-flight mass spectrometry (MALDI-TOF-MS) to obtain peptide mass fingerprinting (PMF) at the protein spots and entered into the NCBI database using Mascot software to identify differentially expressed Of protein. Results: The average protein spots of the nephrotic syndrome hormone-sensitive group and the hormone-resistant group gel were 37 ± 3 and 40 ± 4, respectively; there were differences in urinary protein profiles between the two groups, with 13 differential protein spots; Nine proteins were further identified by 10 protein spots with obvious difference, 4 protein spots were identified by hormone sensitivity group, and 5 protein spots were identified by hormone resistance group. CONCLUSION: Nine different differentially expressed proteins in urine of children with hormone-sensitive and steroid-resistant nephrotic syndrome were screened by two-dimensional electrophoresis and MALDI-TOF-MS.