不同月龄新西兰兔肝中差异蛋白质鉴定与验证

Identification and validation of differential proteins in liver of New Zealand rabbits at different months of age

兔出血症是由兔出血症病毒(Rabbit haemorrhagic disease virus,RHDV)引起的一种兔急性、高度致死性传染病,成年兔死亡率超过90%,但2周龄幼兔不发病。基于幼年兔与成年兔对RHDV的易感性差异,采用串联质谱标记(tadem mass tags,TMT)蛋白质组学技术对幼年兔(2周龄)和成年兔(6月龄)肝中的蛋白质进行检测和定量分析,从幼年兔和成年兔肝中定量获得4353个蛋白质,筛选出821个差异蛋白质;相较于幼年兔,成年兔肝中294个蛋白质显著上调,527个蛋白质显著下调。GO功能富集分析结果显示,生物学过程主要富集到145个小分子代谢蛋白质、111个与氧化还原过程相关的蛋白质和478个参与代谢过程的蛋白质,细胞组分主要富集到528个细胞外成分蛋白质、139个线粒体蛋白质和366个细胞质蛋白质等,分子功能主要富集到342个催化活性蛋白质、93个氧化还原活性蛋白质和50个辅酶结合蛋白质等。821个差异蛋白质在KEGG Pathway数据库中共注释到47条KEGG信号通路,主要涉及代谢途径、糖代谢、氧化磷酸化作用和剪切小体等通路。幼年兔和成年兔肝中差异蛋白质互作网络分析发现,MRPS15的关联度最高,且差异蛋白质胆固醇酰基转移酶1(ACAT1)在互作网络中具有更多相互作用关系。从蛋白质水平探讨幼年兔和成年兔的感染差异机制,筛选并分析幼年兔和成年兔肝中的差异蛋白质,筛选出的差异蛋白质有望成为RHDV体外扩增细胞系构建的突破点,为构建适宜RHDV体外扩增的细胞系提供新的研究思路。

Rabbit hemorrhagic disease is an acute and highly fatal infectious disease caused by Rabbit hemorrhagic disease virus(RHDV).The mortality rate of adult rabbits is more than 90%,but 2-week-old young rabbits are not affected.Based on the differences in the susceptibility of young rabbits and adult rabbits to RHDV,tandem mass tage(TMT)proteomics technology was used to detect and quantitatively analyze the proteins in liver of young rabbits(2 weeks old)and adult rabbits(6 months old).4353 proteins were quantitatively obtained from the liver of young rabbits and adult rabbits,and 821 differential proteins were screened.Among them,294 proteins were significantly up-regulated and 527 proteins were significantly down-regulated in the liver of adult rabbits compared with young rabbits.The results of GO functional enrichment analysis showed that 145 small molecule metabolic proteins,111 redox-related proteins and 478 proteins involved in metabolic processes were mainly enriched in biological processes;528 extracellular component proteins,139 mitochondrial proteins and 366 cytoplasmic proteins were mainly enriched in cellular components;and 342 catalytic active proteins,93 redox active proteins and 50 coenzyme binding proteins were mainly enriched in molecular functions.A total of 47 KEGG(Kyoto Encyclopedia of Genes and Genomes)signaling pathways were annotated in 821 liver differential proteins in the KEGG pathway database,mainly involving metabolic pathways,carbon metabolism,oxidative phosphorylation and splicing bodies.The analysis of differential protein interaction network in liver of young and adult rabbits showed that MRPS15 had the highest correlation degree,and the differential protein cholesterol acyltransferase 1(ACAT1)had more interaction in the interaction network.To explore the mechanism of infection difference between young and adult rabbits from the protein level,and to screen and analyze the differential proteins in liver of young and adult rabbits,it is expected to become a breakthrough point in the construction of RHDV in vitro amplification cell lines,and provide new research ideas for the construction of cell lines suitable for RHDV in vitro amplification.