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草鱼肾细胞来源的外泌体蛋白质组学分析揭示了其在草鱼呼肠孤病毒感染细胞中的反应过程。

Proteomic analysis of the exosomes secreted from Ctenopharyngodon idellus kidney cells infected with grass carp reovirus reveals their involvement in the cellular responses to viral infection.

机构信息

School of Biology & Basic Medical Science, Soochow University, Suzhou, 215123, China.

National Engineering Laboratory for Modern Silk, Soochow University, Suzhou, 215123, China.

出版信息

Fish Physiol Biochem. 2021 Aug;47(4):857-867. doi: 10.1007/s10695-021-00939-4. Epub 2021 Mar 20.

DOI:10.1007/s10695-021-00939-4
PMID:33745109
Abstract

Exosomes are small membrane-enclosed vesicles secreted by various types of cells. Exosomes not only participate in different physiological processes in cells, but also involve in the cellular responses to viral infection. Grass carp reovirus (GCRV) is a non-enveloped virus with segmented, double-stranded RNA genome. Nowadays, the exact role of exosomes in regulating the life cycle of GCRV infection is still unclear. In this study, the exosomes secreted from Ctenopharyngodon idellus kidney (CIK) cells infected or uninfected with GCRV were isolated, and the differential protein expression profiles were analyzed by proteomic technologies. A total of 1297 proteins were identified in the isolated exosomes. The differentially abundant proteins were further analyzed with functional categories, and numerous important pathways were regulated by exosomes in GCRV-infected CIK cells. These exosomal proteins were estimated to interact with the genes (proteins) of the top 10 most enriched signaling pathways. Furthermore, GW4869 exosome inhibitor suppressed the expression level of VP7 in GCRV-infected cells, suggesting that exosomes play a crucial role in the life cycle of GCRV infection. These findings could shed new lights on understanding the functional roles of exosomes in the cellular responses to GCRV infection.

摘要

外泌体是由各种类型的细胞分泌的小膜包裹的囊泡。外泌体不仅参与细胞内的不同生理过程,还参与细胞对病毒感染的反应。草鱼出血病病毒(GCRV)是一种无包膜的病毒,具有分段的双链 RNA 基因组。如今,外泌体在调节 GCRV 感染的生命周期中的确切作用尚不清楚。在本研究中,从感染或未感染 GCRV 的草鱼肾脏(CIK)细胞中分离出外泌体,并通过蛋白质组学技术分析其差异蛋白表达谱。在分离的外泌体中鉴定出 1297 种蛋白质。进一步对差异丰度蛋白进行功能分类分析,发现外泌体在 GCRV 感染的 CIK 细胞中调节了许多重要的途径。这些外泌体蛋白估计与前 10 个最富集信号通路的基因(蛋白)相互作用。此外,GW4869 外泌体抑制剂抑制了 GCRV 感染细胞中 VP7 的表达水平,表明外泌体在 GCRV 感染的生命周期中发挥着关键作用。这些发现可以为理解外泌体在细胞对 GCRV 感染的反应中的功能作用提供新的思路。

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