Expression Profile Of Long Noncoding RNA In Intestinal Rat Macrophages Treated With Lipopolysaccharide

ObjectiveThis study was designed to analyze the differential expression profiles of long noncoding RNA(lncRNA)in lipopolysaccharide(LPS)-induced rat macrophages compared with untreated groups.Bioinformatic analysis was used for further research on differential expressed mRNA.This research lays the foundation for the preliminary study of lncRNA regulatory mechanisms.Methods1.Six rat intestinal macrophages were isolated and randomly divided into a control group consisting of non-lipopolysaccharide(LPS)intestinal macrophages and an experimental group composed of intestinal macrophages that were treated with LPS(at a concentration of 1 mg/L).Differentially expressed lncRNAs and mRNAs from the control group and the experimental group were identified using microarray profiling(Affymetrix Gene Chip Mouse Transcriptome Array 1.0).Hierarchical clustering was used to analyze the differentially expressed lncRNAs and m RNAs.2.In order to elucidate the role of the differentially expressed genes in biological processes,GO analyses and pathway analyses were performed.Furthermore,lncRNA-mRNA-network characterization was performed to locate core regulatory factors.3.As part of this analysis,we used PCR to confirm the presence of differentially expressed m RNAs and lncRNAs.Results1.In total,we identified 357 differentially expressed lncRNAs,consisting of 245up-regulated and 112 down-regulated lncRNAs.Meanwhile 542 differentially expressed mRNAs was fund with comparison between the non-LPS group and the LPS group,which consisted of187 up-regulated mRNAs and 355 down-regulated mRNAs(fold change > 1.5,P < 0.05).2.GO analyses of differentially expressed m RNAs indicated that the biological functions of up-regulated mRNAs were related to inflammatory response?innate immune response?metabolic process?signal transduction and so on.While the biological functions of down-regulated mRNAs included in metabolic process?cell cycle?apoptotic process?inflammatory response.Pathway analyses of differentially expressed mRNAs indicated that the significance pathway of up-regulated mRNAs were related to NF-kappa B signaling pathway?B cell receptor signalingpathway ? Apoptosis and so on.While the pathway of down-regulated mRNAs included in Metabolic pathway ? Phosphatidylinositol signaling system ? Cytokine-cytokine receptor interaction ? Toll-like receptor signaling pathway.In lncRNA-mRNA co-expression network,lncRNA NONMMUT062258?NONMMUT046238?NONMMUT024673?NONMMUT062258may be core factors.3.A significant correlation was subsequently identified between the microarray data and PCR results.ConclusionsThis was the first study to detect the expression profile of lncRNA in rat macrophages treated with LPS by gene chip technique.LncRNAs were significantly differentially expressed in LPS treated intestinal macrophages compared with non-LPS treated intestinal macrophages of rats.PCR verification results showed that the microarray data was stable and reliable.The differentially expressed lncRNAs may regulate the inflammatory response of LPS-stimulated intestinal macrophages of rats.In addition,bioinformatics analysis explored potential regulatory mechanisms.Although more research is needed to demonstrate the exact regulatory mechanisms of these lncRNAs,the genomic data,that we have identified,can provide new clues and ideas for further elucidating the molecular mechanisms of intestinal macrophages regulating inflammatory responses in intestinal barrier dysfunction.