College of Life Sciences and Medicine, Zhejiang Sci-Tech University, Hangzhou 310018, China.
Shaoxing Biomedical Research Institute, Zhejiang Sci-Tech University, Shaoxing 312000, China.
Viruses. 2021 Jun 24;13(7):1218. doi: 10.3390/v13071218.
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 ( < 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.
猪流行性腹泻病毒（PEDV）是引起猪流行性腹泻的病原体。为了研究 PEDV 的致病机制，将 PEDV 接种到体外培养的 Vero 细胞中，提取 Vero 细胞的总 RNA，构建 Illumina 高通量测序文库，筛选差异表达基因（<0.05）。随机选择 5 个用于 qRT-PCR 验证分析的差异表达基因，验证结果与转录组测序结果一致。对筛选出的差异表达基因进行京都基因与基因组百科全书（KEGG）信号通路富集分析。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、P53 信号通路、Jak-STAT 信号通路、MAPK 信号通路和免疫炎症。此外，已有报道称 Puma 可促进细胞凋亡，是 P53 依赖性和非依赖性细胞凋亡途径的关键介质。然而，目前尚无报道称 PEDV 感染可激活 Puma 并诱导 P53 依赖性途径中的细胞凋亡。通过流式细胞术发现 PEDV 感染可诱导细胞凋亡，通过 Western Blotting 检测发现，PEDV 感染可显著增加 p53、BAX 和 Puma 凋亡相关蛋白的表达。用 p53 抑制剂 PFT-α 处理 Vero 细胞可显著抑制 PEDV 诱导的细胞凋亡。研究表明，PEDV 感染可激活 Puma 并诱导 P53 依赖性途径中的细胞凋亡。这些发现为进一步阐明 PEDV 的致病机制和开发有效的 PEDV 疫苗提供了数据支持。
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