【摘要】 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。更多还原

【Abstract】 Objective:To detect urinary polypeptide patterns in stages of the type 2 diabetic nephropathy(DN) in order to screen and find out the potential biomarkers.Methods:A total of 68 urine samples from type 2 diabetic patients with normoalbuminuria(DM1,n = 24),microalbuminuria(DM2,n = 22)and macroalbuminuria(DM3,n = 22),and healthy controls(C,n = 20) were condensed by magnetic beads based weak cation exchange chromatography(MB-WCX),and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF-MS).ClinProtTM software was used to profile and screen the polypeptide patterns in urine of the patients.Results:When the relative molecular mass < 12 000,there were significant differences in 15 protein peaks between group DM1 and group C(P < 0.01).There was one protein peak of significantly low expression in the group DM2,when compared to group C(P = 0.029 8).And there were significant differences in 10 protein peaks between group DM3 and group C(P < 0.01).Conclusion:MB-WCX coupled to MALDI-TOF-MS is a fast,convenient and high throughput analyzing method capable of screening some relatively specific and potential biomarkers from the urine of DN with different stages.更多还原