Abstract
Lipopolysaccharide (LPS) is a component of the cell wall of Gram-negative bacteria, and triggers an inflammatory response both in vitro and in vivo. Here, we used LPS from Escherichia coli serotype enteritidis to stimulate chicken macrophages (HD11) and conducted the transcriptome analysis using a bioinformatics approach to explore the functions of immune-related genes and miRNAs. In total, 1759 differentially expressed genes (DEGs) and 18 differentially expressed (DE)-miRNAs were detected during LPS infection. At 6 h post infection, 1025 DEGs and 10 miRNAs were up-regulated, and 734 DEGs and 8 DE-miRNAs were down-regulated. Based on both RNA hybrid and miRanda systems, 55 DEGs could be targeted by 14 DE-miRNAs. The target genes were related to the immune response, such as IRF8, STAT3, TRAF7, and other potential candidate genes. The DE-miRNAs miR146a-3p, miR6583-5p, and miR30c-2-3p were investigated further. They were predicted to target 34 genes that may also be candidates for immune-related miRNAs and genes. Our results enhanced our understanding of the pathogenic mechanisms of Gram-negative bacteria in chickens.
Highlights
LPS, known as endotoxin, is a highly acylated glycolipid on the cell surface of Gram-negative bacteria
We performed gene ontology (GO) analysis of the miRNA target genes, and 418 terms were significantly enriched and 23 terms were related to immunological processes (p < 0.05; Supplementary Materials Table S7). 29 target genes were predicted to participate in immunization activities
1378 differentially expressed (DE)-mRNAs and 18 miRNAs were identified. 235 genes may be related to immunological processes and LPS-related responses
Summary
LPS (lipopolysaccharide), known as endotoxin, is a highly acylated glycolipid on the cell surface of Gram-negative bacteria. LPS is composed of three parts, a highly acylated di-glucosamine backbone (lipid A) connected to a polysaccharide containing repeating sugars (O-antigen) and linked through a highly conserved oligosaccharide Kdo/heptose core [1,2]. Lipid A can represent its physiological role, and is considered a manifestation of the toxic effects of Gram-negative bacterial infection [3]. Work on LPS began in the 1960s on Escherichia coli and Salmonella typhimurium. LPS is recognized as a primary component of the cell wall and the primary cause of the host cell infection induced by Gram-negative bacteria. LPS has been used in models of the pathogenesis of infections with Salmonella and Escherichia coli
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