基于非依赖性全扫描采集蛋白组学技术初步研究继发性细粒棘球蚴病小鼠肝脏差异表达蛋白

Preliminary study on differentially expressed proteins in a mouse model of secondary cystic echinococcosis based on data independent acquisition proteomics

目的寻找细粒棘球蚴病(CE)小鼠肝脏差异表达蛋白,为细粒棘球蚴病靶向治疗药物研发提供借鉴。方法8只6~8周龄昆明种雌性小鼠随机分成CE组和对照组,CE组小鼠经腹腔注射接种约2000个细粒棘球绦虫原头节,对照组小鼠注射等量生理盐水,饲养350 d后处死两组小鼠。收集CE组小鼠病灶肝脏(病灶组)、病灶旁肝脏组织(病旁组)及对照组小鼠肝脏组织(对照组),采用非依赖性全扫描采集(data independent acquisition,DIA)蛋白组学技术比较病灶组与对照组、病旁组与对照组差异表达蛋白,并进行蛋白基因本体论(GO)富集分析、京都基因和基因组百科全书(KEGG)通路分析。结果病灶组与对照组、病旁组与对照组间同时存在26种差异表达蛋白,其中8种上调表达蛋白、18种下调表达蛋白。GO功能富集注释分析结果显示,这些差异表达蛋白主要富集在内质网膜等细胞内成分中,通过氧化还原酶实现催化等分子功能,参与氧酸代谢、羟基酸代谢等生物代谢过程。KEGG通路富集分析结果显示,酰基辅酶A氧化酶1(Acox1)参与脂肪酸氧化过程中的初级胆汁酸生物合成,影响过氧化物酶体信号通路;肝脏型脂肪酸结合蛋白(Fabp1)通过脂肪细胞因子信号通路参与脂肪酸转运,影响脂质代谢所参与的过氧化物酶体增殖激活受体(PPAR)信号通路。结论与正常小鼠相比,CE小鼠肝脏组织蛋白质表达具有差异性,其中部分差异表达蛋白可能与CE潜在药物靶点相关。

Objective To identify the differentially expressed proteins in different liver tissues in the mouse model of cystic echinococcosis(CE), so as to provide insights into the research and development of therapeutic drugs targeting CE. Methods Female Kunming mice at ages of 6 to 8 weeks were randomly assigned into the CE group and the control group. Mice in the CE group were intraperitoneally infected with 2 000 Echinococcus multilocularis protoscoleces, while mice in the control group were injected with the same volume of physiological saline. All mice in both groups were sacrificed after breeding for 350 d, and the lesions(the lesion group) and peri-lesion specimens(the peri-lesion group) were sampled from the liver of mice in the CE group and the normal liver specimens(the normal group) were sampled from mice in the control group for data independent acquisition(DIA) proteomics analysis, and the differentially expressed proteins were subjected to Gene Ontology(GO) term enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analysis. Results A total of 26 differentially expressed proteins were identified between the lesion group and the normal group and between the peri-lesion group and the normal group, including 8 up-regulated proteins and 18 down-regulated proteins. GO term enrichment analysis showed that these differentially expressed proteins were predominantly enriched in endoplasmic reticulum membrane(biological components), oxidoreductase activity(molecular function) and oxoacid metabolic process and monocarboxylic acid metabolic process(biological processes). KEGG pathway enrichment analysis revealed that the differentially expressed protein Acyl-CoA oxidase 1(Acox1),which contributed to primary bile acid biosynthesis during the fatty acid oxidation, was involved in peroxisome signaling pathway,and the differentially expressed protein fatty acid binding protein 1(Fabp1), which contributed to fatty acid transport, was involved in the peroxisome proliferator-activated receptor(PPAR) signaling pathway. Conclusion Differentially expressed proteins are identified in the liver specimens between mouse models of CE and normal mice, and some differentially expressed proteins may serve as potential drug targets for CE.