| 于 洋,罗福龙,胡迎春,陈睦虎,伍长学.基于蛋白组学的脓毒症补体凝血级联通路及标志物KLKB1研究[J].,2023,(2):377-383 |
| 基于蛋白组学的脓毒症补体凝血级联通路及标志物KLKB1研究 |
| Study on Complement Coagulation Cascade Pathway and Marker KLKB1 in Sepsis Based on Proteomics |
| 投稿时间：2022-05-23 修订日期：2022-06-19 |
| DOI：10.13241/j.cnki.pmb.2023.02.033 |
| 中文关键词: 脓毒症 蛋白组学 补体与凝血级联通路 KLKB1 |
| 英文关键词: Sepsis Proteomics Complement and coagulation cascade pathway KLKB1 |
| 基金项目:四川省科技厅项目（2019JDPT0003；2020YFS0517） |
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| 中文摘要: |
| 摘要 目的：通过蛋白质组学方法鉴定脓毒症关键通路及诊断标志物。方法：选取2019年1月至12月西南医科大学附属医院急诊科收治的56例脓毒症患者（脓毒症组），另取同期50名健康体检志愿者（对照组）。采用随机抽样法分别选取两组中12名脓毒症患者和8名健康体检志愿者，利用非数据依赖模式（DIA）进行血清蛋白数据采集，将数据上传至iDEP在线平台分析脓毒症患者外周血中差异表达蛋白，进一步对这些差异蛋白进行生物信息学分析，包括主成分分析（PCA）、基因本体富集分析（GO）、通路富集分析和蛋白-蛋白相互作用网络（PPI）分析，进而筛选出脓毒症关键蛋白。采用酶联免疫吸附试验（ELISA）对脓毒症组、对照组进行关键蛋白表达验证分析。采用受试者工作特征（ROC）曲线分析关键蛋白对脓毒症的诊断效能。结果：蛋白质组学分析共鉴定出690个蛋白，筛选出171个差异表达蛋白（DEPs），其中39个蛋白显著下调，132个蛋白显著上调。DEPs富集的核心通路为补体和凝血级联通路。该条通路中的血清激肽释放酶 1（KLKB1）在脓毒症组的表达水平为（121.80±55.63 ng/mL），显著高于对照组的（68.30±57.11 ng/mL），差异具有统计学意义（t=4.881，P=0.000）。根据ELISA结果进行脓毒症诊断ROC曲线分析得出，KLKB1蛋白诊断脓毒症的 AUC（95%CI）为0.759（0.594～0.923）。结论：补体和凝血级联通路为脓毒症免疫途径的重要通路，KLKB1具有较好的脓毒症诊断特性，可能是脓毒症潜在的诊断生物标志物。 |
| 英文摘要: |
| ABSTRACT Objective: Identification of key pathways and diagnostic markers of sepsis by proteomics. Methods: 56 patients with sepsis who were treated in the emergency department of the Affiliated Hospital of Southwest Medical University from January to December 2019 were selected as the sepsis group, another 50 healthy volunteers in the same period were taken as the control group, 12 patients with sepsis and 8 healthy volunteers in the two groups were selected by random sampling. Serum protein data were collected by data-independent acquisition (DIA). Data were uploaded to iDEP online platform to analyze differentially expressed proteins in peripheral blood of patients with sepsis. Further bioinformatics analysis was carried out on these differential proteins, including principal component analysis (PCA), gene ontology enrichment analysis (GO), pathway enrichment analysis and protein-protein interaction network (PPI) analysis, so as to screen the key proteins of sepsis, verify them by enzyme-linked immunosorbent assay (ELISA) and analyze the receiver operating characteristic (ROC) curve of sepsis diagnosis. Results: A total of 690 proteins were identified by proteomic analysis, and 171 differentially expressed proteins(DEPs) were screened, of which 39 proteins were significantly down-regulated, and 132 proteins were significantly up-regulated. The core pathway of DEPs enrichment was the link between complement and coagulation cascade pathway. The ELISA results of serum kallikrein 1(KLKB1) in this pathway showed that the expression level of KLKB1 in sepsis group (121.80± 55.63 ng/mL) was significantly higher than that in control group(68.30±57.11 ng/mL), and the difference was statistically significant (t=4.881, P=0.000). According to the results of ELISA, the AUC curve of sepsis diagnosis was 0.759 (95%CI: 0.594-0.923, P=0.002). Conclusion: Complement and coagulation cascade pathways are important immune pathways of sepsis, and KLKB1 has good diagnostic characteristics of sepsis, which may be a potential diagnostic biomarker of sepsis. |
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