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目的 :利用弱阳离子磁珠纯化(magnetic beads based weak cation exchange chromatography,MB-WCX)系统结合基质辅助激光解析电离-飞行时间质谱(matrix-assisted laser desorption/ionization time-of-flight mass spectrometry,MALDI-TOF-MS)的方法获取糖尿病肾病(diabetic nephropathy,DN)的尿小分子差异多肽谱。方法:按Mogensen标准,根据3 d平均微量白蛋白尿排泄率,将68例2型糖尿病患者分组,其中正常蛋白尿(DM1)组24例,微量白蛋白(DM2)组22例,大量蛋白(DM3)组22例;健康对照(C)组20例。DM1及DM2组肾小球率过滤(glomerular filtration rate,GFR)均>90 ml/(min.1.73 m2),DM3组GFR为60～89 ml/(min.1.73 m2)。采用MB-WCX试剂盒富集尿多肽,再经MALDI-TOF-MS技术,采集尿液多肽谱,应用ClinProtTM软件进行生物学比较分析。结果:质荷比<12 000时,DM1组与C组相比,有15个蛋白质峰差异有统计学意义(P<0.01)。DM2组与C组相比,有1个蛋白质峰差异有统计学意义(P=0.029 8)。DM3组与C组相比,有10个蛋白质峰差异有统计学意义(P<0.01)。结论:MB-WCX系统联合MALDI-TOF-MS蛋白质检测技术,能够获得健康人及DN患者尿液小分子多肽谱,为进一步从中寻找早期筛查的标志物提供基础。
OBJECTIVE: To investigate the effect of matrix-assisted laser desorption / ionization time-of-flight mass spectrometry (MALDI-PCR) on magnetic beads based weak cation exchange chromatography (MB- TOF-MS) method was used to obtain the urine small molecule differential peptide profile of diabetic nephropathy (DN). Methods: According to the Mogensen standard, according to the 3-day average microalbuminuria excretion rate, 68 type 2 diabetic patients were divided into groups, including 24 cases of normal proteinuria (DM1), 22 cases of microalbumin (DM2) 22 cases in DM3 group and 20 cases in healthy control group (C). The glomerular filtration rate (GFR) of both DM1 and DM2 groups was> 90 ml / (min.1.73 m2), while that of DM3 group was 60-89 ml / (min.1.73 m2). Urine peptides were enriched by MB-WCX kit and urine peptide profiles were collected by MALDI-TOF-MS. ClinProtTM software was used for biological comparative analysis. Results: When the mass-to-charge ratio <12 000, 15 protein peaks in DM1 group were significantly different from those in C group (P <0.01). Compared with group C, there was a significant difference in protein peak between group DM2 and group C (P = 0.029 8). There were 10 protein peaks in DM3 group compared with C group (P <0.01). Conclusion: MB-WCX system combined with MALDI-TOF-MS protein detection technology can obtain the small molecule peptide profiles of urine in healthy and DN patients and provide a basis for further searching for markers of early screening.