摘要
目的研究伴与不伴糖尿病肾病(DN)的糖尿病视网膜病变(DR)患者血浆蛋白质组学,寻找差异表达蛋白,为筛选相关血浆标志物提供依据。方法入组16例T2DM患者,8例DR伴DN患者纳入DNDR组,8例单纯DR患者纳入DR组,利用蛋白质组非标记(label-free)定量技术对16例血浆标本进行检测,并行液相串联质谱分析(LC-MS/MS),计算蛋白质表达丰度,筛选差异蛋白并行生物信息学分析,包括基因本体(GO)和代谢与信号转导通路(Pathway)注释和富集分析、差异蛋白相互作用网络分析。ELISA法检测两组患者血浆IGFBP7、CPS1 水平,验证label-free 结果。结果以均数和中位数差异倍数均≥ 2.5 倍(上/下调)为标准,筛选出差异蛋白17个,上调11个,下调6个。GO富集分析显示结合蛋白在分子功能蛋白中占绝对优势。Pathway 富集分析示CPS1在丙氨酸,天冬氨酸和谷氨酸代谢中起作用。蛋白相互作用网络分析显示HSPG2、CPS1、IGFBP7、TLN1、KRT5之间有相互作用。ELISA法结果显示IGFBP7在DR组较DNDR组表达升高,CPS1在DR组较DNDR组表达降低。结论采用label-free定量技术发现DR组与DNDR组存在多种差异表达蛋白,这些差异蛋白提供了不同的候选相关血浆标志物和早期防治的靶点。
Objective To study the plasma proteomics of diabetic retinopathy(DR)patients with/without diabetic nephropathy(DN),and to search for differential proteins as to provide a basis for screening related plasma markers.Methods Eight patients with DR and DN were enrolled in the DNDR group.Eight patients with simple DR were enrolled in the DR group.Plasma samples from 16 patients were subjected to protein-labeled-free quantification and parallel liquid-phase tandem mass spectrometry(LC-MS/MS)in order to calculate the protein abundance.The results were statistically analyzed to find significant differences between the two groups of proteins for bioinformatics analysis.These include gene ontology(GO),GO and pathway enrichment analysis,and protein interaction network analysis.The plasma levels of IGFBP7 and CPS1 were measured by ELISA to verify the label-free results.Results A total of 17 differential proteins were screened using mean and median ratios ≥ 2.5(up/down).Among them,11 were up-regulated and 6 were down-regulated.GO enrichment analysis showed that binding protein in the molecular function was dominant.Pathway enrichment analysis showed that CPS1 played a role in the metabolism of alanine,aspartic acid and glutamic acid.The protein interaction network analysis showed that HSPG2,CPS1,IGFBP7,TLN1,KRT5 had interaction.The results of ELISA showed that the expression of IGFBP7 protein in the DR group was higher than that of the DNDR group,and the expression of CPS1 protein was lower in the DNDR group.Conclusion There are many differential proteins in plasma proteomes of patients in DR and DNDR groups by label-free quantification technology.These differential proteins provide different candidate plasma markers and targets for early prevention and treatment.
作者
李志琛
刘颖
何徐军
梅伟群
陈建斌
张华北
欧阳建
钱佳丽
LI Zhichen;LIU Ying;HE Xujun;MEI Weiqun;CHEN Jianbin;ZHANG Huabei;OUYANG Jian;QIAN Jiali(Department of Endocrinology,903 Hospital of PLA,Hangzhou 310004,China)
出处
《实用医学杂志》
CAS
北大核心
2019年第13期2094-2098,共5页
The Journal of Practical Medicine
基金
南京军区医药卫生重点项目(编号:11Z038)
浙江省医药卫生科技计划项目(编号:2013KYA177)
杭州市科技发展计划医药卫生及重点专科专病科研攻关专项(编号:20130633B30)
