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目的利用磁珠分离系统结合基质辅助激光解析电离飞行时间质谱(MALDI-TOF-MS)的方法获取糖尿病肾病(DN)早期的尿小分子差异多肽谱。方法按Mogensen标准,46例2型糖尿病患者经测3 d平均微量白蛋白尿排泄率(UAER)后分组,其中正常蛋白尿组(DM1)24例,微量白蛋白组(DM2)22例,健康对照组(C)20例。DM1及DM2组肾小球率过滤(GFR)均大于90 ml.min-1.1.73m-2。所有受试者留取空腹晨尿经采用弱阳离子(WCX)磁珠纯化试剂盒富集尿多肽,再采用MALDI-TOF-MS技术,获得尿液多肽谱,用ClinProtTM软件进行生物学比较分析。结果相对分子质量<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(均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<0.05)。DM1组与DM2组相比,有10个蛋白质峰差异有统计学意义(均P<0.01)。结论弱阳离子磁珠分离系统联合MALDI-TOF-MS蛋白质检测技术,能够获得健康人及DN早期尿液多肽谱,为进一步从中寻找早期诊断及评估药物干预效果需要的标志物提供前期的基础。
OBJECTIVE: To obtain the differential urinary micro-peptide profiles of early diabetic nephropathy (DN) by using magnetic bead separation system combined with matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS). Methods According to Mogensen criteria, 46 patients with type 2 diabetes underwent 3-day mean microalbuminuria excretion rate (UAER), including 24 cases of normal proteinuria group (DM1), 22 cases of microalbumin group (DM2), and healthy Control group (C) 20 cases. Glomerular filtration rate (GFR) in DM1 and DM2 groups was greater than 90 ml.min-1.1.73m-2. Urine peptides were enriched in all the subjects with fasting morning urine through urinary cation exchange (WCX) purification kit. The urine peptides were obtained by MALDI-TOF-MS. ClinProtTM software was used for biological comparative analysis. Results When the relative molecular mass was less than 12 000, 15 protein peaks in DM1 group were significantly different from those in C group (all P <0.01). There was a significant difference in protein peak between DM2 group and C group (P <0.05). Compared with DM2 group, 10 protein peaks in DM1 group had significant difference (all P <0.01). Conclusions Weak cation magnetic bead separation system combined with MALDI-TOF-MS protein detection technology can obtain the urine peptide profile of healthy people and DN early urine and provide a preliminary basis for further searching for the markers needed for early diagnosis and evaluation of drug intervention effects.