糖尿病肾病差异蛋白质组分析
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Proteome analysis of diabetic nephropathy urinary
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    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-

    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.

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洪 侃,郁志明,石 维,刘志华,许宇静,卞坤娟,黄玉政,徐 明.糖尿病肾病差异蛋白质组分析[J].南京医科大学学报(自然科学版),2012,(7):1033-1037

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  • 收稿日期:2012-01-09
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