原发性痛经气滞血瘀证患者血浆蛋白质组学研究

Plasma proteomics study on primary dysmenorrhea patients with qi stagnation and blood stasis syndrome

目的:从蛋白质表达水平探讨原发性痛经气滞血瘀证的生物学基础。方法:选择符合纳排标准的原发性痛经患者,按中医辨证分型分为气滞血瘀证组、寒凝血瘀证组,另选择健康女性作为正常组,每组8例。采用Label-free定量蛋白质组学对各组月经周期第2天的血浆样本进行分析,获取气滞血瘀证的差异表达蛋白,并进行后续生物信息学分析。结果:气滞血瘀证组与正常组筛选出67个差异表达蛋白;寒凝血瘀证组与正常组筛选出270个差异表达蛋白。经过交集映射获取气滞血瘀证差异表达蛋白47个,其中27个上调蛋白,20个下调蛋白。GO富集分析显示上调蛋白主要集中于免疫反应、代谢等过程;下调蛋白主要集中在代谢、缺氧反应等过程。蛋白结构域富集和聚类显示上调蛋白主要涉及肽聚糖结合、基质素等结构域;下调蛋白主要涉及金属肽酶家族、胰蛋白酶结构域。KEGG富集和聚类显示上调蛋白主要参与嘌呤代谢等通路。蛋白互作网络及拓扑参数分析显示中性粒细胞胶原酶(MMP8)、基质金属蛋白酶9(MMP9)、金属蛋白酶抑制剂2(TIMP2)、乳转铁蛋白(LTF)、E3泛素蛋白连接酶RBX1(RBX1)为核心差异表达蛋白。结论:气滞血瘀证的发生机制主要涉及免疫调节、炎症反应、血管内皮功能障碍、代谢紊乱、缺氧反应等生物学过程和信号通路。利用蛋白质组学寻找的5个核心差异表达蛋白可能作为气滞血瘀证的生物学标志物,对揭示气滞血瘀证的部分实质具有重要意义。

Objective:To explore the biological basis of primary dysmenorrhea with qi stagnation and blood stasis syndrome from protein expression level.Methods:The patients with primary dysmenorrhea meeting the criteria of inclusion and exclusion were selected and divided into qi stagnation and blood stasis syndrome group and cold coagulation and blood stasis syndrome group according to the syndrome differentiation of TCM.In addition,healthy women were selected as the normal group,with 8 cases in each group.The plasma samples of each group on the second day of menstrual cycle were analyzed by Labelfree quantitative proteomics.The differentially expressed proteins of qi stagnation and blood stasis syndrome were obtained and analyzed by bioinformatics.Results:A total of 67 differentially expressed proteins were screened between qi stagnation and blood stasis syndrome group and normal group.A total of 270 differentially expressed proteins were screened between cold coagulation and blood stasis syndrome group and normal group.A total of 47 differentially expressed proteins of qi stagnation and blood stasis syndrome were obtained by overlapped,including 27 up-regulated proteins and 20 down-regulated proteins.GO enrichment analysis showed that up-regulated proteins were mainly concentrated in processes such as immune response,metabolism and down-regulated proteins were mainly concentrated in processes such as metabolism and hypoxia.Protein domain enrichment and clustering analyses showed that the up-regulated proteins were mainly related to peptidoglycan binding and matrixin.The downregulated proteins were mainly involved in metallopeptidase family and trypsin.KEGG enrichment and clustering analyses showed that up-regulated proteins were mainly involved in pathways such as purine metabolism.Protein-protein interaction network and topology parameters analysis showed that neutrophil collagenase(MMP8),matrix metalloproteinase 9(MMP9),metalloproteinase inhibitor 2(TIMP2),lacttransferrin(LTF),E3 ubiquitin protein ligase RBX1(RBX1)were the core differentially expressed proteins.Conclusion:The pathogenesis of qi stagnation and blood stasis syndrome mainly involves biological processes and signal pathways such as immune regulation,inflammatory response,vascular endothelial dysfunction,metabolic disorder,hypoxia response,etc..The five core differentially expressed proteins found by proteomics may be used as biological markers of qi stagnation and blood stasis syndrome,which are of great significance to reveal part of the essence of qi stagnation and blood stasis syndrome.