Analysis Of Differential Expression Of LncRNAs In Piglet Intestinal Inflammation Induced By Lipopolysaccharide

Lipopolysaccharides（LPS）can induce an inflammatory immune responses in the intestine,and long non-coding RNAs（lncRNAs）are involved in the process of inflammatory diseases.However,the biological role of lncRNAs in the intestinal inflammation of piglets is still unclear.In this paper,weaned piglets were stimulated by LPS to establish an intestinal injury inflammation model,and high-throughput sequencing was used to analyze the transcriptome of the ileal mucosa,revealing the expression of lncRNAs in the piglet intestinal inflammation induced by LPS;furthermore,bioinformatics was used to predict the target genes of the differentially expressed candidate lncRNAs and analyze their function enrichment to preliminarily clarify the function of lncRNAs in LPS-induced intestinal inflammation in piglets.In this study,six 35-day-old Du×Long×Large three-way cross-weaned piglets,weighing 9-10 kg,were randomly divided into control group and LPS group,with 3replicates in each treatment.The weaned piglets in the LPS group were injected intraperitoneally with 100μg/kg body weight of LPS,and the weaned piglets in the control group were injected intraperitoneally with the same amount of 0.9%Na Cl solution.Three hours after the injection,the ileal mucosal tissue was collected and used the Illumina Hi Seq 2000 for RNA sequencing.The results showed that compared with the control group,112 lncRNAs were significantly differentially expressed in the ileal mucosa of the piglets treated with LPS(|log₂^FC|≥1,P<0.05),of which 58 lncRNAs were significantly up-regulated（P<0.05）,54 lncRNAs were significantly down-regulated（P<0.05）;in addition,in the LPS group,there were 415 mRNAs with significant differential expression(|log₂^FC|≥1,P<0.05),of which 228 mRNAs were significantly up-regulated（P<0.05）and 187 mRNAs were significantly down-regulated（P<0.05）.The target genes of differentially expressed lncRNAs were predicted and analyzed by Gene Ontology and Kyoto Encyclopaedia of Genes and Genomes The results indicate that these target genes are mainly involved in the signaling pathways related to inflammatory immune response or injury,such as cell adhesion molecules（CAMs）signaling pathways and m TOR signaling pathways.In addition,Cyto Scape software was used to construct a co-expression network diagram between differentially expressed lncRNAs and mRNA（|Pearson correlation coefficient|>0.9,P<0.05）,and identify 7 core lncRNAs（SLA-8,ITGA10,WWC3,SIGLEC1,TMP-SLA-5,TMP-SLA-3 and PDGFRA）.The 7 core lncRNAs were verified by real time-quantitative PCR,and the results showed that lncRNA TMP-SLA-3,TMP-SLA-5,SLA-8,SIGLEC1,ITGA10,WWC3 were all significantly up-regulated in the LPS-induced ileal mucosa（P<0.05）;lncRNA PDGFRA was significantly down-regulated in LPS-induced ileal mucosa（P<0.01）.It is speculated that the interaction between these differentially expressed lncRNAs and their target genes may have an important impact on the process of LPS-induced inflammation.In conclusion,the differentially expressed lncRNAs in LPS-induced intestinal inflammation response of piglets were studied in this study.After bioinformatics prediction and functional analysis,7 core lncRNAs were excavated,which were speculated to be involved in the regulation of intestinal inflammation,providing a good research basis for the study of the function of lncRNAs in LPS-induced intestinal inflammation.