miRNA-125b regulates TNF-α production in CD14+ neonatal monocytes via post-transcriptional regulation

Neonates, although deficient in cell immunity, frequently reveal sepsis with augmented proinflammatory reactions. Here, we found that neonatal monocytes produced significantly higher TNF-alpha mRNA and protein than adult monocytes. Assessment of the transcriptional factor found no significant difference of NF-kappa B p65 level between neonatal and adult monocytes. Addition of Act D to access the half-life of TNF-alpha mRNA revealed no significant difference of the LPS-induced TNF-alpha mRNA half-life between them, whereas CHX increased neonatal TNF-alpha mRNA significantly. This suggests that a post-transcriptional mechanism involves the augmentation of TNF-alpha production by neonatal monocytes. To examine whether miRNA was involved in the post-transcriptional regulation, differential displays of miRNA array between neonatal and adult MNCs were performed, along with the discovery of hsa-miR-103, hsa-miR-125b, hsa-miR-130a, hsa-miR-454-3p, and hsa-miR-542-3p, which were greater than a twofold decrease or increase after LPS treatment for 4 h. The functional validation identified that miR-125b decreased significantly in association with higher TNF-alpha expression by neonatal monocytes after LPS stimulation. Transfection of the miR-125b precursor into neonatal monocytes significantly repressed the TNF-alpha mRNA and protein expression, suggesting that miR-125b negatively regulates TNF-alpha expression in neonatal monocytes. Modulation of miRNA expression may be used to regulate TNF-alpha production in newborns with altered proinflammatory reactions. J. Leukoc. Biol. 92: 171-182; 2012.