基于转录组和同位素标记相对与绝对定量技术的宽体金线蛭生品和烫制品差异蛋白研究  

Differential protein study between raw and processed products of Whitmania pigra Whitman using transcriptome and isobaric tags for relative and absolute quantitation proteome

作　　者：华羽彤 李尹 郭秀欢 雷艳 辛泉诚 袁瑞娟[1] HUA Yu-tong;LI Yin;GUO Xiu-huan;LEI Yan;XIN Quan-cheng;YUAN Rui-juan(Beijing University of Chinese Medicine,Beijing 102488,China)

机构地区：[1]北京中医药大学,北京102488

出　　处：《中国新药杂志》2024年第3期292-300,共9页Chinese Journal of New Drugs

基　　金：国家自然科学基金资助项目(82173955)。

摘　　要：目的:通过转录组和蛋白质组技术研究宽体金线蛭生品和炮制品的差异蛋白。方法:以活体宽体金线蛭(Whitmania pigra Whitman)为原料,基于无参转录组进行基因分析,利用转录组信息匹配到Helobdella robusta蛋白质库,然后利用同位素标记相对与绝对定量技术(iTRAQ)蛋白组学技术进行宽体金线蛭的蛋白成分分析,筛选出存在于宽体金线蛭生品及炮制品中的差异蛋白质,并对其进行基因本体(GO)功能聚类分析、京都基因与基因组百科全书(KEGG)富集分析以及蛋白质相互作用网络分析(PPI),并结合BIOPEP数据库筛选可能具有抗凝活性的蛋白。结果:本研究筛选到差异表达蛋白共112种,其中78种上调蛋白、34种下调蛋白,并推定出6条可能具有抗凝作用的差异蛋白。GO功能聚类分析结果表明,在细胞过程(cellular process)、代谢过程(metabolic process)条目富集的差异蛋白数量最多。在差异性方面,大分子复合物(macromolecular complex)、结构分子活性(structural molecule activity)条目差异最显著。KEGG富集分析结果表明,差异蛋白在代谢途径(metabolic pathways)富集到的蛋白最多,ECM-受体相互作用(ECM-receptor interaction)、核糖体(ribosome)、氧化磷酸化(oxidative phosphorylation)差异最显著。结论:本研究结合转录组、蛋白组数据,补充了宽体金线蛭的蛋白质库,筛选出了生品及烫制品的差异蛋白并推定了可能具有抗凝作用的差异蛋白,为宽体金线蛭基因组成、物质基础研究及炮制机制提供参考依据。Objective:To study the differential proteins between raw and processed products of Whitmania pigra Whitman by transcriptome and proteome techniques.Methods:Using Whitmania pigra Whitman as a raw material,genetic analysis was carried out based on the parameter-free transcriptome,and then transcriptome information was matched to the Helobdella robusta protein library.The protein composition of Whitmania pigra Whitman was analyzed by isobaric tags for relative and absolute quantitation(iTRAQ)proteomics technology,and the differential proteins presented in the raw and processed products of Whitmania pigra Whitman were screened out.We also analyzed the functional clustering of gene ontology(GO),the Kyoto Encyclopedia of Genes and Genomes(KEGG),and protein-protein interaction network analysis(PPI).We also screened the proteins with possible anticoagulant activity using the BIOPEP database.Results:A total of 112 differentially expressed proteins were screened out in this study,including 78 up-regulated proteins and 34 down-regulated proteins.Finally,6 differential proteins that have potential anticoagulant activity were presumed.The results of GO functional clustering analysis showed that the largest number of differential proteins were enriched in the entries of cellular process and metabolic process.In terms of differences,the entries of macromolecular complex and structural molecular activity were the most significant.The results of KEGG enrichment analysis showed that differential proteins were enriched in the metabolic pathways,and the differences in ECM-receptor interaction,ribosome,and oxidative phosphorylation were the most significant.Conclusion:In this study,the transcriptome and proteome data were combined to supplement the protein library of Whitmania pigra Whitman,and the differential proteins of raw and processed products were screened.The differential proteins that have potential anticoagulant activity were presumed,which provided a reference for the genetic composition,material basis and the processing m

关 键 词：宽体金线蛭 炮制 转录组 同位素标记相对与绝对定量技术蛋白组 差异蛋白 

分 类 号：R917[医药卫生—药物分析学]