The Changes Of Expression Profiles And Bioinformatics Analysis Of LncRNAs In Macrophages Under Simulated Microgravity

ObjectiveTo study the differential expression of long non-coding RNAs?LncRNAs?in macrophages under simulated microgravity and explore the biological significance.MethodsThe RAW264.7 macrophages were randomly divided into control and simulated microgravity groups,the cells in simulated microgravity were treated by clinostat for 72 hours.The profile of LncRNAs were detected with Arraystar mouse lncRNA chips and the significant difference expression of LncRNAs were screened out with the difference more than 2 times and statistically significant?P < 0.05?.The differential LncRNAs were verified by qRT-PCR technique.The GO enrichment and KEGG pathway analysis of the target genes from screened LncRNAs were carried out.The inflammatoty factors were detected with multiplexed immunoassay based on Luminex technology from eBioscience and cell apoptosis was measured with flow cytometry.Western blotting were used to detect the changes of related proteins expression.ResultsCompared with control group,the results from LncRNA microarray showed that there were 693 differentially expressed LncRNAs,in which 328 LncRNAs were up-regulated and 365 LncRNAs were down-regulated.qRT-PCR results showed that the expression of LncRNA ENSMUST00000137470,uc009 oqw.1,uc007 wem.2,NR₀15575 and uc.462 were consistent with LncRNA chip results.GO results showed that differentially expressed LncRNAs were significantly enriched in the biological process including positive regulation of inflammatory response.KEGG results showed that the top 10 significant signal pathways included IL-17,TNF-?,Toll receptors,MAPK pathways and cell apoptosis regulation,etc.The preliminary functional study showed that simulated microgravity induced the increased secretion of inflammatory factors TNF-?,IL-1?,IL-6 and IL-18,increased cell apoptosis.The results from western blotting showed that simulated microgravity can regulate the inflammatory response through MAPK pathway,and take part in the regulation of cell apoptosis through controlling the expression of apoptosis related proteins.ConclusionsSimulated microgravity changed the expression profile of LncRNAs in RAW 264.7 macrophages,the differentially expressed LncRNAs may play the role of cellular physiological regulation by controlling the biological process of inflammatory response and cell apoptosis.