基于DIA定量蛋白组学分析生脉饮对大鼠血浆蛋白表达的影响

Effects of Shengmai Yin on protein expression in plasma of rats based on DIA quantitative proteomics analysis

目的采用数据非依赖型采集(data independent acquisition,DIA)定量蛋白组学技术分析生脉饮灌胃前后大鼠血浆蛋白表达谱的差异,为明确生脉饮对正常个体的作用及机制提供依据。方法取6只大鼠,给予生脉饮0.02ml/(g·d)灌胃,连续5d,灌胃前后采血行DIA定量蛋白组学分析。结果共鉴定到1093个蛋白,筛选出32个上调差异表达蛋白(differentially expressed proteins,DEPs)和35个下调DEPs。京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)数据库注释显示,DEPs主要参与免疫系统等信号通路;基因本体论(Gene Ontology,GO)富集分析显示DEPs主要富集于内源性凝血途径、纤维蛋白凝块形成和急性炎症反应的调节等过程;KEGG富集分析显示DEPs主要富集于补体和凝血级联等信号通路。蛋白互作网络分析显示,上调蛋白中血红素结合蛋白、类黏蛋白1、补体成分4等相互作用蛋白较多,以参与补体和凝血级联以及中性粒细胞胞外诱捕网形成的相关蛋白为主,而下调蛋白中鸟苷三磷酸(guanosine triphosphate,GTP)结合核蛋白相互作用蛋白较多。结论生脉饮可明显改变正常大鼠血浆蛋白表达,其中可能涉及炎症和凝血的功能调节。

Objective To analyze the differences in the plasma protein expression profiles of rats before and after gavage administration of Shengmai Yin using the quantitative proteomics technique of data independent acquisition(DIA),and provide a basis for clarifying the effects and mechanisms of Shengmai Yin on normal individuals.Methods Six rats were administered Shengmai Yin at a dose of 0.02ml/(g·d)by gavage for 5 consecutive days.Blood was collected before and after gavage for DIA quantitative proteomics analysis.Results A total of 1093 proteins were identified,and 32 up-regulated differentially expressed proteins(DEPs)and 35 down-regulated DEPs were screened.The Kyoto Encyclopedia of Genes and Genomes(KEGG)database annotation showed that DEPs were mainly involved in signaling pathways such as the immune system.Gene Ontology(GO)enrichment analysis indicated that DEPs were mainly enriched in processes such as endogenous coagulation pathways,fibrin clot formation,and regulation of acute inflammatory responses.KEGG enrichment analysis displayed that DEPs were mainly enriched in signaling pathways such as complement and coagulation cascade.Protein interaction network analysis displayed that there were more interacting proteins including hemopexin,orosomucoid 1,complement 4 and other proteins in the up-regulated proteins,and they were predominantly related proteins involved in the complement and coagulation cascades as well as in the formation of the extracellular trapping network ofneutrophils,whereas there were more interaction proteins with guanosine triphosphate(GTP)-binding nuclear protein in the down-regulated proteins.Conclusion Shengmai Yin significantly alter the expression of plasma proteins in normal rats,which may be involved in functional regulation of inflammation and coagulation.