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猪肠上皮细胞感染猪流行性腹泻病毒的代谢组学和蛋白质组学分析。

Metabolomic and Proteomic Profiling of Porcine Intestinal Epithelial Cells Infected with Porcine Epidemic Diarrhea Virus.

机构信息

Key Laboratory for Animal Genetics, Breeding, Reproduction and Molecular Design, College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China.

Animal Breeding and Genetics, Graduate School of Agricultural Science, Tohoku University, Sendai 980-8572, Japan.

出版信息

Int J Mol Sci. 2023 Mar 7;24(6):5071. doi: 10.3390/ijms24065071.

Abstract

Porcine epidemic diarrhea virus (PEDV) infection results in severe epidemic diarrhea and the death of suckling pigs. Although new knowledge about the pathogenesis of PEDV has been improved, alterations in metabolic processes and the functional regulators involved in PEDV infection with host cells remain largely unknow. To identify cellular metabolites and proteins related to PEDV pathogenesis, we synergistically investigated the metabolome and proteome profiles of PEDV-infected porcine intestinal epithelial cells by liquid chromatography tandem mass spectrometry and isobaric tags for relative and absolute quantification techniques. We identified 522 differential metabolites in positive and negative ion modes and 295 differentially expressed proteins after PEDV infection. Pathways of cysteine and methionine metabolism, glycine, serine and threonine metabolism, and mineral absorption were significantly enriched by differential metabolites and differentially expressed proteins. The betaine-homocysteine S-methyltransferase (BHMT) was indicated as a potential regulator involved in these metabolic processes. We then knocked down the BHMT gene and observed that down-expression of BHMT obviously decreased copy numbers of PEDV and virus titers ( < 0.01). Our findings provide new insights into the metabolic and proteomic profiles in PEDV-infected host cells and contribute to our further understanding of PEDV pathogenesis.

摘要

猪流行性腹泻病毒(PEDV)感染会导致仔猪严重腹泻和死亡。尽管人们对 PEDV 的发病机制有了新的认识,但宿主细胞中与 PEDV 感染相关的代谢过程和功能调节剂的变化在很大程度上仍不清楚。为了鉴定与 PEDV 发病机制相关的细胞代谢物和蛋白质,我们通过液相色谱串联质谱和等重标记相对和绝对定量技术,协同研究了 PEDV 感染猪肠上皮细胞的代谢组和蛋白质组图谱。我们在正离子和负离子模式下鉴定出 522 个差异代谢物和 295 个 PEDV 感染后的差异表达蛋白。差异代谢物和差异表达蛋白显著富集了半胱氨酸和蛋氨酸代谢、甘氨酸、丝氨酸和苏氨酸代谢以及矿物质吸收途径。甜菜碱-同型半胱氨酸 S-甲基转移酶(BHMT)被认为是参与这些代谢过程的潜在调节剂。随后,我们敲低了 BHMT 基因,观察到 BHMT 的下调明显降低了 PEDV 的拷贝数和病毒滴度(<0.01)。我们的研究结果为 PEDV 感染宿主细胞的代谢和蛋白质组图谱提供了新的见解,并有助于我们进一步了解 PEDV 的发病机制。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0c34/10049511/16223b051c1c/ijms-24-05071-g001.jpg

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