?-3 Polyunsaturated Fatty Acid Regulates The Expression Of MicroRNAs In Dendritic Cells And Bioinformatics Prediction

Objective:Dendritic cells are important antigen presenting cells playing important roles in innate immunity and acquired immunity.In the inflammatory bowel disease and graft-versus-host response,DC activation plays a key role.Omega-3 polyunsaturated fatty acids can play an immunosuppressive effect by inhibiting the maturation of dendritic cells and inflammatory response and inducing immune tolerance.microRNAs,as an important gene regulatory molecule,can affect dendritic cells maturation and antigen presentation process.Omega-3 polyunsaturated fatty acids can affect the expression of multiple microRNAs in a variety kinds of cells and can also influence the expression profile of microRNAs in peripheral blood of human or other animals.It is involved in regulating multiple signal pathways and cellular functions.Omega-3 polyunsaturated fatty acids may influence the maturation and antigen presenting functions of dendritic cells throughing multiple cell functions and signaling pathways by affecting microRNAs expression profile of dendritic cells.Methods:1.The immature dendritic cells were obtained through 7 days of mouse bone marrow precursor cells culture using GM-CSF and IL-4.After dendritic cells were treated with Omega-3 polyunsaturated fatty acids for 24 hours,lipopolysaccharide was added to make them mature,and then they were collected for cell experiments;2.The effect of omega-3 polyunsaturated fatty acids on the phosphorylation of p-38 in dendritic cells was detected by western blotting;3.The effects of ?-3 polyunsaturated fatty acids on the surface costimulatory molecules and adhesion molecules CD80,CD83 and CD86 of dendritic cells were detected by flow cytometry;4.The effects of omega-3 polyunsaturated fatty acids on the levels of IL-6,IL-10,IL-12 and TNF-? secretion by dendritic cells were detected by ELISA;5.Dendritic cells were treated with omega-3 polyunsaturated fatty acids and the total RNA was extracted with TRIZOL method.The microRNAs expression profile of each group was detected by RNA microarray.EPA group and DHA group were compared with the mature group to find the microRNAs expression profile difference;6.microRNAs with significant changing were detected using gene microarray and were verified by taqman probe method;7.Through the gene ontology analysis(GO analysis)and KEGG pathway analysis,we found the cellular functions,targeting proteins and signaling pathways regulated by microRNAs.Results:1.Compared with the mature group,omega-3 polyunsaturated fatty acids changed the microRNAs expression profile of dendritic cells.2.Proteins,targeted by omega-3 polyunsaturated fatty acids changed microRNAs of dendritic cells mainly concentrated in MAPK,PI3K/Akt,Notch and other signal pathways.3.The expression of CD80 and CD86 on dendritic cells was down-regulated by omega-3 polyunsaturated fatty acids,but the expression of CD83 was not affected;4.Compared with the mature group,omega-3 polyunsaturated fatty acids can inhibit the p-38 phosphorylation of dendritic cells;5.Omega-3 polyunsaturated fatty acids inhibited the secretion of IL-6,IL-12 and TNF-? of dendritic cells;Conclusion:1.?-3 can affect the maturation of dendritic cells;2.omega-3 polyunsaturated fatty acids may change the expression profile of microRNAs and proteins targeted by microRNAs,which are mainly concentrated in the MAPK,PI3K/Akt signaling pathway.