miR-485-5p regulates the inflammatory response of macrophages by targeting interleukin-1 receptor-associated kinase 4
Objective This paper aims to investigate the targeting effect of miR-485-5p on interleukin-1 receptor associat-ed kinase 4(IRAK-4)and its regulatory role in the inflammatory response of macrophage model.Methods miR-485-5p mimics and their inhibitors were synthesized and transfected into HEK-293T cells.The expression of miR-485-5p was detected by RT-PCR.IRAK-4 was selected as the target gene for the study according to the analysis of miRNA targeting effect prediction data,and IRAK-4 3UTR and its mutant firefly-Renilla dual luciferase reporter gene system were construc-ted.The targeting relationship between miR-485-5p and IRAK-4 was verified by detecting the effect of overexpression/inhi-bition of miR-485-5p on luciferase activity in the cell model,and the protein expression of IRAK-4 was detected by West-ern blot.The macrophage model was constructed,and miR-485-5p mimics and inhibitors were transfected into macrophages,respectively.The expression of target gene IRAK-4 mRNA was detected by RT-PCR,and the expression of pathogenic factors IL-6,IL-17 and IL-18 in the supernatant was detected by enzyme-linked immunosorbent assay(ELISA).Results It was confirmed by experiments that miR-485-5p and IRAK-4 had a targeted effect.In the cell model,compared with the control group,the expression of miR-485-5p after transfection of miR-485-5p mimics increased by 17.15 times(P＜0.01),and the expression of miR-485-5p after transfection of miR-485-5p inhibitors decreased by 79.80％(P＜0.05).Over-expression of miR-485-5p significantly inhibited the protein level and mRNA relative content of IRAK-4 and the expression of downstream inflammatory factors,IL-6,IL-17 and IL-18(P＜0.05).Inhibiting the expression of miR-485-5p up-regulated the protein level and relative mRNA content of IRAK-4 and significantly increased the release of IL-6,IL-17,and IL-18(P＜0.05).Conclusion miR-485-5p can regulate the expression of downstream inflammatory cytokines by targeting the specific sequence of IRAK-4,and then regulate the inflammatory response of cells.
miR-485-5pInterleukin-1 receptor associated kinase 4Human leukemia monocytic cell line
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NSTL
万方数据
维普
