急性脑出血瘀热病机单元的差异蛋白质组学研究

Differential Proteomics Analysis of Pathogenic Unit of Stasis-heat for Acute Intracerebral Hemorrhage

目的明确急性脑出血瘀热病机单元的生物标志物及其病理生理机制。方法收集急性脑出血患者271例,参照《瘀热病机证候量化诊断标准评分表》划分为瘀热病机组与非瘀热病机组并进行1∶1配对,两组各选取4例患者,采集每例患者的血浆样本,采用串联质谱标签法(TMT)进行定量蛋白质组检测,取两组间蛋白质表达量>1.3倍且P≤0.05的蛋白质为差异蛋白质。应用创新途径分析(IPA)在线生物信息学软件对差异蛋白质进行经典通路、上游调控因子、分子与细胞功能及蛋白质相互作用网络分析。结果每例患者共鉴定出490种蛋白质,其中9种蛋白质表达两组存在差异:A1BG、ACTN1、CA1、CD44、CP、PRDX2、VCL、IgG H chain、IGHV3-7,IPA软件识别出7种(A1BG、ACTN1、CA1、CD44、CP、PRDX2、VCL),瘀热病机组与非瘀热病机组比较,CD44表达下调,其余6种蛋白表达上调,主要涉及5条经典信号传导通路、2个上游调控因子、5种分子与细胞功能;生成了1个确信度较高的差异蛋白质生物学作用网络。结论差异蛋白质参与了PI3K复合物、Akt和ERK相关的细胞信号通路网络的调节,这些通路与炎症因子及凝血指标密切有关或是其调节的上游通路。

Objective To clarify the biomarkers and molecular disorder mechanism of pathogenic unit of stasis-heat of acute intracerebral hemorrhage. Methods A total of 271 acute cerebral hemorrhage patients and their plasma samples were collected and classified into stasis-heat type and non-stasis-heat type according to Quantitative Diagnostic Scale of Stasis-heat Syndrome. Then 4 of each group were selected by 1∶1 paired design, and plasma samples of 4 pairs were collected or detecting differential proteins by tandem mass tag(TMT). The proteins expressed more than 1.3 times and P≤0.05 between the two groups were taken as differential proteins. The Ingenuity Pathway Analysis(IPA) bioinformatics software was performed in analyzing canonical pathways, upstream regulatory factor, molecular cellular functions, and protein interaction network of differential proteins. Results A total of 490 proteins were identified and quantified in each patient, of which 9 proteins were differentially expressed between two groups(A1BG, ACTN1, CA1, CD44, CP, PRDX2, VCL, IgG H chain, and IGHV3-7), and A1BG, ACTN1, CA1, CD44, CP, PRDX2, and VCL were identified by IPA software. Compared with non-stasis-heat group, CD44 was down-regulated and the remaining six proteins were up-regulated. These proteins involved 5 canonical signaling pathways, 2 upstream regulators and 5 kinds of molecules and cell functions. One associated functional network was generated with acceptable confidence. Conclusion Differential proteins,reflecting the potential biomarkers of pathogenic unit of stasis-heat of the acute cerebral hemorrhage, are involved in the regulation of PI3K complexes, Akt and ERK-related cellular signaling pathways, which are closely related to inflammatory factors or their regulated upstream pathways.