Des(rhamnosyl) verbascoside抗HBV作用的定量蛋白质组学研究

Quantitative Proteomics Study of Anti-HBV Effects of Des（rhamnosyl） verbascoside

关键词:  des(rhamnosyl) verbascoside  蛋白组学  乙型肝炎病毒  生物信息学  旱田草

Key words:  des(rhamnosyl) verbascoside  proteomics  HBV  bioinformatics  Lindernia ruellioides (Colsm.) Pennell

摘要:

为探究des(rhamnosyl) verbascoside体外抗乙型肝炎病毒(HBV)的活性作用和机制，本研究以des(rhamnosyl) verbascoside为实验药物对HepG2.2.15细胞进行干预，实验分为药物干预组和对照组，采用串联质谱标签(Tandem Mass Tag, TMT)蛋白质组学方法对提取的总蛋白进行分析。结果表明，共筛选得到300个差异表达蛋白，其中有109个上调蛋白，191个下调蛋白。基因本体论(Gene Ontology, GO)分析结果显示，差异蛋白主要参与DNA复制(DNA replication)、鞘糖脂代谢(Glycosphingolipid metabolic process)、细胞增殖(Cell proliferation)、寡糖分解代谢(Oligosaccharide catabolic process)等生物学过程，以及DNA聚合酶活性(DNA polymerase activity)、丝氨酸型羧肽酶活性(Serine-type carboxypeptidase activity)、DNA引物酶活性(DNA primase activity)等分子功能。京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes, KEGG)分析结果显示，差异蛋白主要参与细胞代谢(Metabolism)、遗传信息传导(Genetic information processing)、生物系统通路(Organismal systems)等相关信号通路。亚细胞定位分析表明，差异蛋白大多定位在细胞质和细胞核。本研究共筛选出13个与抗HBV密切相关的蛋白。通过定量蛋白组学初步揭示des(rhamnosyl) verbascoside可能通过增加HGF、SORT1、MAN2B1,减少PRIM1、PRIM2、POLA1、POLD3、POLD2、POLD1、POLE、ERCC2、LAMC1、SDC1等蛋白表达来起到体外抗HBV的作用。

Abstract:

In order to explore the activity and mechanism of des(rhamnosyl) verbascoside against hepatitis B virus (HBV) in vitro,the des(rhamnosyl) verbascoside was used as an experimental drug to intervene HepG2.2.15 cells.The experiment was divided into drug intervention group and control group.Tandem Mass Tag (TMT) proteomics method was used to analyze the extracted total protein.The results showed that a total of 300 differentially expressed proteins were screened,of which 109 proteins were up-regulated and 191 proteins were down-regulated.Gene Ontology (GO) analysis showed that the differentially expressed proteins were mainly involved in DNA replication,glycosphingolipid metabolic process,cell proliferation,and oligosaccharide catabolic process and other biological processes,as well as DNA polymerase activity,serine-type carboxypeptidase activity,DNA primase activity and other molecular functions.The results of Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that the differentially expressed proteins were mainly involved in metabolism,genetic information processing,and organismal systems and other related signaling pathways.Subcellular localization analysis showed that most of the differentially expressed proteins were localized in the cytoplasm and nucleus.A total of 13 proteins closely related to anti-HBV were screened in this study.Through quantitative proteomics,it is preliminarily revealed that des (rhamnosyl) verbascoside may play an anti-HBV role in vitro by increasing HGF,SORT1,MAN2B1 and reducing the expression of PRIM1,PRIM2,POLA1,POLD3,POLD2,POLD1,POLE,ERCC2,LAMC1,SDC1 and other proteins.