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妊娠母猪血清蛋白促进非洲猪瘟病毒复制:一种基于 iTRAQ 的定量蛋白质组学分析。

Proteins in pregnant swine serum promote the African swine fever virus replication: an iTRAQ-based quantitative proteomic analysis.

机构信息

State Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou University, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, 730000, China.

出版信息

Virol J. 2023 Mar 28;20(1):54. doi: 10.1186/s12985-023-02004-3.

Abstract

African swine fever (ASF) is a severe infectious disease caused by the African swine fever virus (ASFV), seriously endangering the global pig industry. ASFV possesses a large genome, strong mutation ability, and complex immune escape mechanisms. Since the first case of ASF was reported in China in August 2018, it has had a significant impact on social economy and food safety. In the present study, pregnant swine serum (PSS) was found to promote viral replication; differentially expressed proteins (DEPs) in PSS were screened and identified using the isobaric tags for relative and absolute quantitation technology and compared with those in non-pregnant swine serum (NPSS). The DEPs were analyzed using Gene Ontology functional annotation, Kyoto Protocol Encyclopedia of Genes and Genome pathway enrichment, and protein-protein interaction networks. In addition, the DEPs were validated via western blot and RT-qPCR experiments. And the 342 of DEPs were identified in bone marrow-derived macrophages cultured with PSS compared with the NPSS. The 256 were upregulated and 86 of DEPs were downregulated. The primary biological functions of these DEPs involved signaling pathways that regulate cellular immune responses, growth cycles, and metabolism-related pathways. An overexpression experiment showed that the PCNA could promote ASFV replication whereas MASP1 and BST2 could inhibit it. These results further indicated that some protein molecules in PSS were involved in the regulation of ASFV replication. In the present study, the role of PSS in ASFV replication was analyzed using proteomics, and the study will be provided a basis for future detailed research on the pathogenic mechanism and host interactions of ASFV as well as new insights for the development of small-molecule compounds to inhibit ASFV.

摘要

非洲猪瘟(ASF)是由非洲猪瘟病毒(ASFV)引起的一种严重传染病,严重威胁全球养猪业。ASFV 具有庞大的基因组、强大的突变能力和复杂的免疫逃避机制。自 2018 年 8 月中国首次报告 ASF 病例以来,它对社会经济和食品安全产生了重大影响。本研究发现,怀孕母猪血清(PSS)可促进病毒复制;使用同位素标记相对和绝对定量技术筛选和鉴定 PSS 中的差异表达蛋白(DEPs),并与非怀孕母猪血清(NPSS)中的 DEPs 进行比较。通过基因本体论功能注释、京都基因与基因组百科全书通路富集以及蛋白质-蛋白质相互作用网络对 DEPs 进行分析。此外,还通过 Western blot 和 RT-qPCR 实验对 DEPs 进行了验证。与 NPSS 相比,在骨髓来源的巨噬细胞中培养 PSS 时鉴定出 342 个 DEPs。其中 256 个上调,86 个下调。这些 DEPs 的主要生物学功能涉及调节细胞免疫反应、生长周期和代谢相关途径的信号通路。过表达实验表明,PCNA 可促进 ASFV 复制,而 MASP1 和 BST2 可抑制其复制。这些结果进一步表明,PSS 中的一些蛋白分子参与了 ASFV 复制的调控。本研究通过蛋白质组学分析了 PSS 在 ASFV 复制中的作用,为进一步研究 ASFV 的致病机制和宿主相互作用以及开发抑制 ASFV 的小分子化合物提供了依据。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ee16/10044734/a34202c4bd57/12985_2023_2004_Fig1_HTML.jpg

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