Fujian Agriculture and Forestry University, Fuzhou, China.
Hainan University, Haikou, China.
PeerJ. 2023 Jun 7;11:e15459. doi: 10.7717/peerj.15459. eCollection 2023.
The intestinal epithelium barrier serves as a highly dynamic immunologic frontier in the defense against invading pathogenic bacteria and viruses. Hence, understanding of the complicated underlying relationship between enteric pathogens and the intestinal epithelium barrier is vital for developing strategies to improve the intestinal health of farm animals. To this end, Caco-2 cells were stimulated by 1 µg/ml lipopolysaccharide (LPS) for 24 h and 5 µg/ml polyinosinic-polycytidylic acid (ploy(I:C)) for 4 h to imitate bacterial and viral infection processes, respectively. The specific alterations in gene expression of Caco-2 cells after stimulation were characterized by transcriptome sequencing. Seventy differentially expressed genes (DEGs) were identified under LPS exposure, and 17 DEGs were observed under ploy(I:C) exposure. We found that most DEGs were specific, and only one common DEG was observed. Gene Ontology (GO) annotation analysis indicated that all DEGs identified in the different treatments were mainly derived from GO terms related to the maintenance of cellular homeostasis. Moreover, specific DEGs such as , , and regulated by LPS treatment, while and mediated by ploy(I:C) treatment, which are derived from immune function modulation related GO terms, were confirmed by both transcriptome sequencing and qRT-PCR. In addition, both transcriptome sequencing and qRT-PCR results verified that LPS specifically down-regulated the DEGs and , which are involved in inflammation responses related to the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway including the TGF-beta signaling pathways and the Ras signaling pathway. Ploy(I:C) uniquely suppressed the DEGs and , which participated in viral replication-associated pathways including autophagy and mTOR signaling pathway.
肠上皮屏障作为抵抗入侵的致病性细菌和病毒的高度动态免疫前沿,对于开发改善农场动物肠道健康的策略至关重要。为此,我们用 1 µg/ml 的脂多糖(LPS)刺激 Caco-2 细胞 24 h,用 5 µg/ml 的聚肌胞苷酸(ploy(I:C))刺激 4 h,分别模拟细菌和病毒感染过程,然后用转录组测序来描述 LPS 和 ploy(I:C)刺激后 Caco-2 细胞基因表达的特定变化。在 LPS 暴露下鉴定出 70 个差异表达基因(DEGs),ploy(I:C) 暴露下鉴定出 17 个 DEGs。我们发现大多数 DEGs 是特异性的,只有一个共同的 DEG。基因本体论(GO)注释分析表明,不同处理中鉴定出的所有 DEGs 主要来源于与细胞内稳态维持相关的 GO 术语。此外,通过 LPS 处理调节的特异性 DEGs ,而通过 ploy(I:C)处理调节的 ,来源于与免疫功能调节相关的 GO 术语,这两个基因都通过转录组测序和 qRT-PCR 得到了证实。此外,转录组测序和 qRT-PCR 结果均验证了 LPS 特异性下调了参与炎症反应相关的 KEGG 通路(包括 TGF-β信号通路和 Ras 信号通路)的 DEGs ,而上调了参与病毒复制相关通路(包括自噬和 mTOR 信号通路)的 DEGs 。ploy(I:C) 则特异性抑制了参与病毒复制相关通路(包括自噬和 mTOR 信号通路)的 DEGs 。