Analysis of Differentially Expressed Genes in the Spleen of Sepsis Mice Based on RNA-Seq
LU Liyue,YONG Yue,GAO Hao
Department of Anesthesiology, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China
Abstract:
Objective To detect the changes and characteristics of mRNA expression in the spleen of sepsis mice induced by LPS and look for the signaling pathways that affect the pathophysiological process of sepsis. Methods The mice were divided into LPS and Sham group, and the mortality and serum levels of TNF-α, IL-6 and IL-1β were observed to evaluate the model. Extract RNA from the spleens of mice, RNA-Seq to determine gene expression and then analyze by two methods: Differentially expressed genes (P<0.05, |log2 Fold Change|≥1) were screened. GO and KEGG were performed and Enrichment of genes with P<0.05 and Q<0.25 in GSEA v4.0.1 software. RT-qPCR verified the 10 differentially expressed genes in the RNA-Seq results. Results LPS successfully constructed a sepsis model, which showed severe inflammation and high mortality (P<0.01). Compared with the Sham group, 1616 differentially expressed genes were changed in the LPS group, of which 919 were down-regulated genes and 697 were up-regulated genes. GO and KEGG analysis involve multiple pathways of inflammation, regulation of genetic information, signal transduction, cellular processes.The RT-qPCR verification results showed that the expression trends of 10 genes were consistent with the sequencing results. Conclusion The cytoplasmic DNA sensing pathway may play an important role in the pathological process of sepsis. Zbp1, Ddx58, Cxcl10 and Polr1d may be important targets for their functions.
