Research Paper Volume 12, Issue 10 pp 9224—9239

Apelin enhances IL-1β expression in human synovial fibroblasts by inhibiting miR-144-3p through the PI3K and ERK pathways

Figure 5. APLN-induced suppression of miRNA-144-3p enhances IL-1β production. (A) Open-source software (TargetScan, miRMap, RNAhybrid, and miRWalk) was used to identify which miRNAs could possibly interfere with IL-1β transcription. (B) OASFs were incubated with APLN (0, 1, 3, and 10 ng/mL). Levels of miR-144-3p expression were examined by RT-qPCR assay (n=4). (C, D) OASFs were transfected with miR-144-3p mimic and then stimulated with APLN (10 ng/mL). mRNA and excreted protein levels were examined by RT-qPCR (n=4) and ELISA assays (n=5). (E) OASFs were transfected with the mut-IL-1β-3′UTR plasmid with or without miRNA-144-3p mimic, then stimulated with APLN (10 ng/mL). Relative luciferase activity reflected IL-1β promoter activity (n=6). (F) OASFs were treated with PI3K or ERK inhibitor then incubated with APLN. miR-144-3p expression levels were examined by RT-qPCR assay (n=4). Results are expressed as the mean ± S.E.M. * p<0.05 compared with the control group; # p<0.05 compared with the APLN-treated group.