磁珠分离结合生物质谱分析肾小球疾病患者尿液多肽谱  

Determination of urinary polypeptide patterns of glomerulonephritis by magnetic bead separation and MALDI-TOF MS

作　　者：吴杰[1] 李燕[2] 谢院生[1] 王杰[2] 王娜[2] 尹忠[1] 侯凯[1] 陈香美[1] 

机构地区：[1]解放军总医院解放军肾脏病研究所暨重点实验室,北京100853 [2]国家生物医学分析中心

出　　处：《中华肾脏病杂志》2009年第8期596-600,共5页Chinese Journal of Nephrology

基　　金：国家自然科学基金重点项目（30630033）;国家科技支撑计划（2007BA104810）

摘　　要：目的利用磁珠分离系统联合基质辅助激光解析电离-飞行时间质谱（MALDI—TOFMS）探讨人体尿液多肽谱，期望找到肾小球疾病诊断相关标记物。方法利用磁珠分离系统富集尿多肽，再由MALDI—TOFMS采集健康对照组29例和肾小球疾病组34例（IgA肾病10例，膜性。肾病10例，微小病变9例，狼疮肾炎5例）患者的尿液多肽谱，用SVM算法和ClinProt^TM软件进行生物学比较分析。结果两组性别、年龄差异无统计学意义。相对分子质量〈10000时，健康对照组平均出峰85个，疾病组平均出峰109个。疾病组与健康对照组相比，有32个蛋白质峰差异有统计学意义（P〈0．01），峰度表达差异在2倍以上者有14个，其中表达上调有6个峰，相对分子质量分别为3371．5、4026．35、4085．32、4116．96、4126．32、9527．31；表达下调有8个峰，相对分子质量分别为861．28、1205．41、1642．52、1913．15、1976．52、2087．74、2193．47、3015．57。疾病组中IgA肾病、膜性肾病、微小病变、狼疮。肾炎均呈现各自特异的尿液多肽谱图。结论磁珠分离系统联合MALDI—TOFMS蛋白质检测技术，能够获得健康人及肾小球疾病患者尿液多肽谱，可能因此得到肾小球疾病相对特异的潜在标志物，可以为临床诊断及研究提供便捷无创的途径。Objective To determine the urinary polypeptide patterns of glomerulonephritis by magnetic bead separation and matrix-assisted laser desorption/ionization timeof-flight mass spectrometry （MALDI-TOF MS） technology. Methods Ufinatry samples of 29 healthy volunteers and 34 patients with glomerulonephritis, including 10 cases of IgA nephropathy （IgAN）, 10 cases of membranous nephropathy （MN）, 9 cases of minimal change disease （MCD） and 5 cases of lupus nephritis （LN）, were collected and separated by magnetic bead,and were screened for polypeptide patterns with a novel high throughput method, MALDI-TOF MS. Results Under the relative molecular weights 10 000 Da, 85 protein peaks were detected in healthy controls group and 109 protein peaks were detected in glomerulonephritis group. Six peaks of 3371.5 Da, 4026.35 Da, 4085.32 Da, 4116.96 Da, 4126.32 Da and 9527.31 Da were up-regulated,while 8 peaks of 861.28 Da, 1205.41 Da, 1642.52 Da, 1913.15 Da, 1976.52 Da, 2087.74 Da, 2193.47Da and 3015.57 Da were down-regulated by more than 2 folds （P〈0.01） in glomerulonephritis group as compared to healthy controls. Urinary polypeptide patterns in different diseases differed significantly from each other, indicating specific disease pattern of polypeptide excretion. Conclusions MALDI-TOF MS is a fast, convenient and high throughput analyzing method capable of screening some relative specific, potential biomarkers from the urine of glomerulonephritis patients thus it possesses better clinical value.

关 键 词：质谱分析法 肾小球肾炎 尿液 多肽 磁珠分离 基质辅 助激光解析电离一飞行时间质谱 

分 类 号：R692.6[医药卫生—泌尿科学] R446.122[医药卫生—外科学]