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Porcine Epidemic Diarrhea Virus Induces Vero Cell Apoptosis via the p53-PUMA Signaling Pathway

机译:猪流行性腹泻病毒通过P53-Puma信号通路诱导Vero细胞凋亡

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摘要

Porcine Epidemic Diarrhea Virus (PEDV) is the causative agent of swine epidemic diarrhea. In order to study the pathogenic mechanism of PEDV, PEDV was inoculated into Vero cells cultured in vitro, and the total RNA of Vero cells was extracted to construct a library for Illumina high-throughput sequencing and screening of differentially expressed genes (p < 0.05). Five differentially expressed genes for qRT-PCR verification analysis were randomly selected, and the verification results were consistent with the transcriptome sequencing results. The Kyoto Encyclopedia of Genes and Genomes (KEGG) signal pathway enrichment analysis was performed on the differentially expressed genes screened above. The results showed that the target gene annotations of differentially expressed genes in the African green monkey genome were mainly enriched in the TNF signaling pathway, the P53 signaling pathway, the Jak-STAT signaling pathway, the MAPK signaling pathway, and immune inflammation. In addition, it has been reported that Puma can promote apoptosis and is a key mediator of P53-dependent and non-dependent apoptosis pathways. However, there is no report that PEDV infection can activate Puma and induce apoptosis in a P53-dependent pathway. It was found by flow cytometry that PEDV infection induced apoptosis, and by Western Blotting detection, PEDV infection significantly increased the expression of p53, BAX, and Puma apoptosis-related proteins. Treatment Vero cells with the p53 inhibitor, PFT-α, could significantly inhibit PEDV-induced apoptosis. Studies have shown that PEDV infection can activate Puma and induce apoptosis in a P53-dependent pathway. These findings provide data support for further elucidating the pathogenic mechanism of PEDV and developing an effective vaccine against PEDV.
机译:猪流行性腹泻病毒(PEDV)是猪流行性腹泻的致病剂。为了研究PEDV的致病机理,将PEDV接种到体外培养的Vero细胞中,并萃取Vero细胞的总RNA,构建illumina高通量测序和差异表达基因的筛选文库(P <0.05) 。随机选择五种差异表达的QRT-PCR验证分析的基因,并且验证结果与转录组测序结果一致。对基因和基因组(Kegg)信号途径富集分析的京都百科全书在筛选的差异表达基因上进行。结果表明,非洲绿猴基因组中差异表达基因的靶基因注释主要富集在TNF信号通路中,P53信号通路,JAK-STAT信号通路,MAPK信号通路和免疫炎症。此外,据报道,PUMA可以促进细胞凋亡,是P53依赖性和非依赖性凋亡途径的关键介质。然而,没有报告,PEDV感染可以激活P53依赖性途径中的露珠并诱导细胞凋亡。通过流式细胞仪发现PEDV感染诱导的细胞凋亡,并通过Western印迹检测,PEDV感染显着增加了P53,Bax和Puma凋亡相关蛋白的表达。治疗Vero细胞与P53抑制剂,PFT-α可以显着抑制PEDV诱导的细胞凋亡。研究表明,PEDV感染可以在P53依赖性途径中激活PUMA并诱导细胞凋亡。这些发现提供了数据支持,用于进一步阐明PEDV的致病机制,并对PEDV发育有效疫苗。

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