首页|非标记定量方法研究糖尿病患者血清差异蛋白

非标记定量方法研究糖尿病患者血清差异蛋白

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目的 应用非标记定量技术研究糖尿病患者血清中差异表达蛋白,筛选特异性蛋白质标志物.方法 收集2023年深圳市糖尿病患者和健康对照者的血清各3例,应用免疫亲和层析和纳升液相色谱-四极杆-静电场轨道-线性离子阱三合一质谱法,利用分级(fraction,FC)和不分级(no fraction,NF)2种方法对糖尿病患者和健康对照者的血清蛋白质组进行非标记定量分析,质谱数据经MaxQuant1.6.1.0进行检索,对分级方法鉴定的差异蛋白质进行GO(gene oncology)功能聚类分析和KEGG(kyoto encyclopedia of genes and genomes)通路分析.结果 未分级方法鉴定可信蛋白251种,筛选差异蛋白17种;分级方法鉴定可信蛋白494种,筛选差异蛋白74种.其中7种蛋白在2种方法中变化趋势完全一致.有5种蛋白下调,分别是妊娠区带蛋白(pregnancy zone protein,PZP)、碳酸酐酶1(carbonic anhydrase 1,CAH1)、甘露糖结合蛋白 C(mannoee-binding protein C,MBL2)、纤维蛋白原a链(fibrinogen alpha chain,FIBA)和触珠蛋白(haptoglobin,HPT);2种蛋白上调,分别是脂蛋白a[apolipoprotein(a),LPA]和补体因子H相关蛋白4(complementfac-tor H-relatedprotein4,CFH4).GO分析显示差异蛋白的分子功能主要集中在糖胺聚糖结合、肝素结合等过程.KEGG通路分析显示差异蛋白主要集中在补体和级联凝血、细胞黏附、PI3K-Akt信号通路等通路上.结论 基于分级的非标记定量蛋白质组学分析技术可有效分析糖尿病患者血清蛋白的差异表达,为筛选大规模队列疾病血清样本的特异性蛋白质标志物提供了有效技术手段.
Research on differentially expressed serum proteins in diabetic patients by label-free quantification proteomics
Objective To investigate the differential expression of proteins in the serum of diabetic patients quantitative proteomics technologies,and to identify specific protein biomarkers.MethodsSerum samples from three diabetic patients and three healthy controls in Shenzhen in 2023 were collected.Immunoprecipitation and nano liquid chromatography-quadrupole-electrostatic orbitrap-linear ion trap mass spectrometry(easy-nanoLC-Q-orbitrap-fusion)were employed to perform fraction and no-fraction-based quantitative analysis of the serum proteome in diabetic patients and healthy controls.The mass spectrometry data were analyzed by MaxQuant1.6.1.0,and the differential proteins identified by the fraction-based method were subjected to gene ontology(GO)functional clustering and kyoto encyclopedia of genes and genomes(KEGG)pathway analysis.Results A total of 251 reliable proteins were identified by the no-fraction-based method,among which 17 differentially expressed proteins were selected.In contrast,494 reliable proteins were identified by the fraction-based method,and 74 differentially expressed proteins were selected.Among them,seven proteins showed consistent changes in both methods.Five proteins were downregulated,including pregnancy zone protein(PZP),carbonic anhydrase 1(CAH1),mannose-binding protein C(MBL2),fibrinogen alpha chain(FIBA),and haptoglobin(HPT).Two proteins were upregulated,including apolipoprotein(a)(LPA)and complement factor H-related protein 4(CFH4).GO analysis revealed that the differentially expressed proteins were mainly involved in molecular functions such as glycosaminoglycan binding and heparin binding.KEGG pathway analysis showed that the differentially expressed proteins were mainly associated with complement and coagulation cascades,cell adhesion,PI3K-Akt signaling pathway,and other pathways.Conclusions Fraction-based no-fractional quantitative proteomics analysis provides an effective approach for analyzing differential protein expression in the serum of diabetic patients,and offers a valuable technique for screening specific protein biomarkers of serum samples in large-scale cohorts of disease.

Label-free quantificationdiabetesserumproteomicsfractionation

刘雅雯、冯里茹、胡绪乔、洪文旭

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汕头大学公共卫生学院,广东汕头 515063

深圳市慢性病防治中心,广东深圳 518020

非标记定量 糖尿病 血清 蛋白组学 分级

深圳医疗卫生"三名工程"项目

SZSM201811057

2024

中国热带医学
中华预防医学会,海南疾病预防控制中心

中国热带医学

CSTPCD北大核心
影响因子:0.722
ISSN:1009-9727
年,卷(期):2024.24(3)
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