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。
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 （DMI,n = 24）,microalbuminuria （DM2,n = 22）and maeroalbuminuria （DM3,rt = 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.
出处
《南京医科大学学报（自然科学版）》
CAS
CSCD
北大核心
2012年第7期1033-1037,共5页
Journal of Nanjing Medical University(Natural Sciences)
关键词
磁珠分离
基质辅助激光解吸电离-飞行时间质谱
糖尿病肾病
尿液
多肽
magnetic beads separation
matrix-assisted laser desorption/ionization time-of-flight mass spectrometry
diabeticnephropathy
urine
polypeptide
