基于蛋白质组学的冠心病痰瘀互结证生物标志物及诊断模型研究  

Biomarkers and Diagnostic Model of Coronary Heart Disease with Intermingled Phlegm and Blood Stasis Syndrome Based on Proteomics

作　　者：杨光[1] 何浩强[1] 周思远 王阶[1] YANG Guang;HE Hao-qiang;ZHOU Si-yuan;WANG Jie(Guang′anmen Hospital,China Academy of Chinese Medical Sciences,Beijing 100053)

机构地区：[1]中国中医科学院广安门医院,北京100053

出　　处：《世界中西医结合杂志》2023年第9期1874-1880,1890,共8页World Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金项目(81974556)。

摘　　要：目的痰瘀互结证是当代冠心病的主要证候之一,而目前尚缺乏可靠的定量蛋白质组学检测结果。基于数据非依赖性采集模式(Data independent acquisition,DIA)对冠心病痰瘀互结证患者的血浆蛋白质组进行研究分析,探索冠心病痰瘀互结证的蛋白标志物及关键生物学过程,并构建证候的诊断模型。方法选取2020年8月—2020年11月期间中国中医科学院广安门医院心血管科收治的5例冠心病痰瘀互结证患者作为疾病观察组,同时选取2020年10月—2020年11月期间在读研究生的健康成人5人作为对照组,采用DIA蛋白质组学技术对受试者的血浆进行检测,进行差异蛋白表达分析及GO富集分析、KEGG富集分析、蛋白质互作分析。根据生物信息学结果筛选冠心病痰瘀互结证的关键蛋白,对其进行酶联免疫吸附试验(Enzyme-linked immunosorbent assay,ELISA),并根据ELISA结果开展logistic回归分析。结果在差异倍数为1.5倍时,疾病观察组相比于健康对照组存在9个蛋白质表达上调,19个蛋白质表达下调。生物信息学分析表明,血管性血友病因子(VWF)、补体因子H是参与凝血和补体反应的关键蛋白。进一步ELISA结果表明,冠心病痰瘀互结证患者的血浆VWF水平高于健康受试者,差异有统计学意义(P<0.05),CFH水平低于健康受试者,差异有统计学意义(P<0.05),与蛋白质组学的结果相一致。VWF的AUC=0.73,CFH的AUC=0.66。基于两指标的Logistic回归模型测试集的准确率、灵敏度、特异度和AUC分别为73%、74%、70%和0.78。结论VWF相较于CFH具有更好的诊断冠心病痰瘀互结证的潜力。基于CFH和VWF两种蛋白质分子的证候诊断模型,具有一定的诊断效能。Objective Intermingled phlegm and blood stasis syndrome is one of the main syndromes in contempo⁃rary coronary heart disease,and reliable quantitative proteomic results are currently lacking.This study utilized data-inde⁃pendent acquisition(DIA)to analyze the plasma proteome of patients with coronary heart disease with intermingled phlegm and blood stasis syndrome to explore protein markers and key biological processes associated with this syndrome and con⁃struct a diagnostic model for the syndrome.Methods Five patients diagnosed with coronary heart disease with intermin⁃gled phlegm and blood stasis syndrome,admitted to the Cardiovascular Department of Guang′anmen Hospital,China Acade⁃my of Chinese Medical Sciences,between August and November 2020,were selected and assigned to the disease observa⁃tion group.Simultaneously,five healthy graduate students from October to November 2020 were selected and assigned to the control group.DIA proteomic technology was employed to analyze plasma samples from the participants.Differential protein expression analysis,GO enrichment analysis,KEGG pathway analysis,and protein-protein interaction analysis were con⁃ducted.Key proteins related to coronary heart disease with intermingled phlegm and blood stasis syndrome were selected based on bioinformatics results.Enzyme-linked immunosorbent assay(ELISA)was performed on selected proteins,and lo⁃gistic regression analysis was conducted based on ELISA results.Results At a fold change threshold of 1.5,the disease observation group exhibited upregulation of nine proteins and downregulation of 19 proteins compared with the control group.Bioinformatics analysis identified von Willebrand factor(VWF)and complement factor H(CFH)as key proteins in⁃volved in coagulation and complement reactions.ELISA results further confirmed that plasma levels of VWF in patients with coronary heart disease with intermingled phlegm and blood stasis syndrome were significantly higher than those in healthy subjects(P<0.05),while CFH levels we

关 键 词：冠心病 痰瘀互结证 蛋白质组学 DIA 证候诊断模型 

分 类 号：R541.4[医药卫生—心血管疾病]