Abstract
A macrophage is an important component of innate immunity which can be activated by infection. A series of inflammatory cytokines are produced and released to eliminate pathogens. CpG DNA is an immune stimulator recognized by TLR9, subsequently inducing inflammatory responses in macrophages. Long noncoding RNA (lncRNA) is a novel class of noncoding RNA, whose length is more than 200 nt, but without protein-coding capacity. lncRNAs are involved in many physiological and pathological processes, including inflammatory responses. In our study, a lncRNA microarray assay was performed to identify differentially expressed lncRNAs and mRNAs in RAW264.7 cells at different time points following CpG ODN stimulation. The results revealed that expression levels of 734 lncRNAs and 734 mRNAs were altered at all time points. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) biological pathway analyses were performed to predict the functions of dysregulated genes. Coexpression networks of lncRNA-mRNA were constructed based on the correlation analysis between differentially expressed lncRNAs and 10 selected upregulated mRNAs, which have been reported to be involved in CpG DNA-induced inflammatory responses. In addition, we selected 8 dysregulated lncRNAs for further validation by quantitative real-time PCR. The present study provided a systematic perspective on the potential functions of lncRNAs in CpG ODN-induced macrophage activation.
Highlights
Innate immunity is the first line of host defense against the pathogen threats [1]
The gene microarray analysis for CpG ODN-activated macrophages was consistent with TLR9 signaling activation, which showed that most inflammatory related-gene expression was upregulated (Figure 1(d))
From the Long noncoding RNA (lncRNA)-mRNA coexpression network, we found that functional molecules and inflammatory cytokines involved in CpG DNA-induced inflammatory response, including Ctsk, Nfkb2, IL-1β, and IL-6, were coexpressed with multiple lncRNAs, which formed a complex network
Summary
Innate immunity is the first line of host defense against the pathogen threats [1]. A macrophage is an important component of innate immunity and plays a crucial role in the inflammatory responses [2]. Recognition of microbial molecules including lipopeptides, lipopolysaccharides, and DNA by pattern recognition receptors of macrophages such as Toll-like receptors (TLRs) will trigger the intracellular signaling cascades. The phagocytosis of macrophages is enhanced to remove the pathogen [2, 3]. Excessive inflammation in macrophages can cause host damage and even lead to chronic inflammatory diseases, such as obesity, cardiovascular disease, inflammatory bowel disease, and cancer [3]. It is required for us to better understand the regulatory mechanisms that limit the excessive inflammatory mediators in macrophages
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