Expression Of Mirnas Of Lung Tissues In Endotoxemic Mouse And Primary Research Of Their Functions

ObjectiveThis study was attempted to detect expression of miRNAs of lung tissues in endotoxemic mouse and investigate the role of miRNAs against endotoxemia.Method1.A animal model of acute lung injury was prepared by injection of E.coli lipopolysaccharide intraperitoneally, and high-throughput miRNA array technology was used to screen miRNAs differentially expressed in lung tissues in endotoxemic mouse.2.QRT-PCR is used to redetect some of these miRNAs to verify if the expression of these miRNAs is consistent with the result of high-throughput miRNA array.3.Expression of miR-1949was detected in RAW264.7cells treated with LPS for different time and with different doses of LPS.4.A software for miRAN target gene miRDB, was used to predict the target genes of miR-1949, then QRT-PCR was used to detect the expression of target genes on the level of tissues and cells after LPS stimulation.Results1.The result of high-throughput miRNA array showed that17miRNAs were upregulated, and9miRNAs were downregulated.2.The expression of miR-1949, miR-223, miR-150was consistent with the result of miRNA array screening. miR-1949and miR-223were upregulated, and miR-150was downregulated.3.Expression of miR-1949in RAW264.7cells after LPS stimulation was time and dose dependent. 4. The target genes of miR-1949we picked out among so many genes were MAPK8, MAP2K1, SMEK1. The expression of SMEK1on the level of tissues and cells after LPS stimulation was downregulated.Conclusion1.The expression of miR-1949in lung tissues and RAW264.7cells after LPS stimulation was upregulated;2.miR-1949may play important role in lung injury in endotoxemic mouse through the expression regulation of SMEK1.