尿液多肽组的时间可变性和相对稳定性研究  

Study on Time Variability and Relative Stability of Urinary Peptidomics

作　　者：万磊磊 陈琦[2] 程思明 李水明 王勇 WAN Lei-Lei;CHEN Qi;CHENG Si-Ming;LI Shui-Ming;WANG Yong(Shenzhen Key Laboratory of Marine Bioresources and Ecology,Brain Disease and Big Data Research Institute,College of Life Sciences and Oceanography,Shenzhen University,Shenzhen 518055,China;The Eighth Hospital Affiliated to Sun Yat-sen University,Shenzhen 518026,China;Shenzhen Bay Laboratory,Shenzhen 518055,China;Shenzhen-Hong Kong Institute of Brain Science-Shenzhen Fundamental Research Institutions,Shenzhen 518055,China)

机构地区：[1]深圳大学生命与海洋科学学院,深圳市海洋生物资源与生态环境重点实验室,深圳市脑病和大数据研究所,深圳518055 [2]中山大学附属第八医院,深圳518026 [3]深圳湾实验室,深圳518055 [4]深圳-香港脑科学研究院,深圳518055

出　　处：《分析化学》2021年第9期1546-1554,共9页Chinese Journal of Analytical Chemistry

基　　金：深圳市科技创新委员会项目(Nos.JCYJ20170818142154551,JCYJ20200109110001818);深港脑科学研究所-深圳基础研究机构项目(No.2021SHIBS0003)资助。

摘　　要：尿液多肽组可反映机体的病理变化,提供潜在的生物标志物信息。目前,尿液多肽组研究通常采用晨尿,其成份是否随采样时间变化尚不清楚。本研究采集了3个健康志愿者的不同时间点的尿液,经氧化石墨烯-磷酸镧纳米复合材料(LaGM)方法分离后,利用LC-MS/MS进行序列鉴定。发现在不同取样时间下,自同一个人的尿液多肽组成分存在差异,降解肽段及其所归属的蛋白质种类和数目具有时间可变性,这种差异在高丰度降解蛋白体现为肽段数目和序列的变化,在低丰度蛋白则表现为“有”或“无”的改变,在24 h内多次取样分析,显著增加了尿液中多肽的检出数目。另一方面,尿液多肽组也具有一定的稳定性,尿调节素的降解肽段SGSVIDQSRVLNLGPITR不仅在各时间点普遍存在,而且相对强度通常高于其序列相关肽段,而胶原蛋白是晨尿中(6时尿液)的优势组分。采用单人单时间点尿液、不同年龄段混合尿液和文献数据对上述特征进行验证分析,结果基本符合。本研究结果表明,尿液多肽组成分随采样时间动态变化,24 h尿液比晨尿更适合作为样本来源,更能全面反映受试者尿液多肽组的情况,而由于阶梯序列的存在和个体差异等因素,肽段的绝对强度不宜作为筛选标志物的依据。All the peptides in the urine is peptidome,which can reflect the pathological changes of the body,thereby providing potential biomarker information.At present,morning urine is most often used in the research of urinary peptidomics.However,whether its composition changes with the sampling time is not clear.In this study,the urine samples from the three healthy people at different time points were separated by the graphene oxide-lanthanum phosphate nanocomposite(LaGM)method and identified by LC-MS/MS.Protein Pilot Software 4.5 was used for database search and analysis.It was found that the peptides and proteins of urine at each time point of the same person were different,which was reflected in the change of number and sequence of peptides in high-abundance degraded proteins and“with”or“without”in low-abundance.The number of peptides detected in urine significantly increased by sampling a plurality of times in a day.On the other hand,the peptidomics was also certain stable.The degradation peptides SGSVEDQSRVLNLGPITR of uromodulin not only existed at every time point but also had higher relative strength than other sequence-related peptides.Collagen was the dominant component in morning urine(urine at 6 o′clock).Further more,the result was verified by single urines,mixed urines of different ages,and literature data.This study showed that the sampling time was the main influencing factor of urinary peptidomics.The 24-h urine samples would be more suitable for sample than the morning urine.Due to the existence of the ladder sequence and the influence of individual differences,the absolute signal intensity of peptide should not be used for direct comparison between different samples and the selection of biomarkers.

关 键 词：晨尿 24 h尿液 多肽组学 质谱 尿调节素 

分 类 号：R446.12[医药卫生—诊断学] O657.63[医药卫生—临床医学]