Long Noncoding RNA And MicroRNA Sequencing Analysis Of Different Time Points Of LPS-treated Mice’s Lung Tissue

Sepsis is the worldwide leading cause of infectious deaths with high fatality rate.Nearly 1000 people died from sepsis per hour.Pathogenesis of sepsis is still unclear.So that the clinically treatment of sepsis isn’t timely with poor prognosis.Lipopolysaccharide(LPS)is the main component of gram-negative bacteria cell walls.LPS is often used to induce sepsis mice model by intraperitoneal injection.High-throughput sequencing technology developing rapidly in recent years.This technology leads a revolusion in scitntific research and gene diagnosis.Scientis can explore biological and medical problems from the perspective of big data for accurate medical escort.Objective:This study applied high-throughput sequencing technologies to explore the dynamic changes of mRNA,lncRNA and microRNA of LPS-induced sepsis mice lung tissue.Method:12 C57BL/6 healthy male mice(9-week-old,20-22 g)were purchased from the Experimental Animal Center of Southern Medical University,and divided into four groups with random number tables:a.The 0 h group(LA),intraperitoneal injection with normal saline(20 ml/kg b.w,0.9%NaCl);b.The 2 h group(LB),2 h after intraperitoneal injection with LPS;c.The 8 h group(LC),8 h after intraperitoneal injection with LPS;d.The 24 h group(LD),24 h after intraperitoneal injection with LPS.The dose of intraperitoneal injection of LPS is 20 mg/kg b.w.Pick the lung tissue from the anesthetized mice at different time points after the perfusion from heart.LncRNA and microRNA of lung tissue were analyzed by high throughput sequencing.Results:(1)The number of gene’s different alternative splicing events was elevated as the extension of LPS stimulation time(747,1442 and 2453 respectively).(2)2638,6453 and 6191 different expression mRNAs were found respectively at the three LPS-treated time points.Using clustering analys,gene ontology enrichment analysis,KEGG pathway enrichment analysis and protein-protein interaction analysis to analyse these different expression mRNAs.(3)175,362,305 different expression lncRNAs and 110,246,188 different expression TUCPs and 10,93,54 different expression miRNAs were found respectively at the three LPS-treated time points.These different expression lncRNAs,TUCPs and miRNAs were clustered and make target genes prediction respectively.Using clustering analysis,gene ontology enrichment analysis,KEGG pathway enrichment analysis and protein-protein interaction analysis to analyse these different expression RNAs’s predicted target genes.Conclusion:(1)Gene’s different alternative splicing involved in mouse lung tissue in respons to LPS treated.(2)There was a significant difference in the number of differentially expressed mRNA in the lungs and liver of mice after LPS stimulation for 24 h,suggesting that these two organs have different compensation under LPS stimulation and thus affect their degree of damage.(3)The enrichment of GO mRNA in lung differentially expressed mRNA was the most obvious in the immune system process and immune response.The enrichment of extracellular vesicular exosome and membrane-bound vesicle at 8 h and 24 h after LPS stimulation suggested that LPS stimulation Late mouse lung cell signal interaction was significantly affected.In addition to the classical TNF signaling pathway,KEGG significantly enriched the NFκB signaling pathway,and the proteasome signaling pathway was highly activated 24 h after LPS stimulation.Find differences in core protein interaction network changes.(4)The number of differentially expressed transcripts was greater than the number of down-regulated at different time points,suggesting that the transcriptional regulation of the mouse mainly by up-regulating the transcripts under LPS stimulation.(5)There was no significant change of sRNA in lung tissue at different time points after LPS stimulation.