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Our results demonstrated a dynamic change of the expressions of miR\u2010149 expression and MyD88 in macrophage RAW264.7 upon <jats:italic>Mycobacterium bovis<\/jats:italic> Bacillus Calmette\u2010Guerlin (BCG) infection or lipopolysaccharide (LPS) stimulation. The presence of BCG or LPS dynamically reduced the miR\u2010149 expression, along with a substantially striking increase of MyD88 expression in these cells. More importantly, overexpression of miR\u2010149 in RAW264.7 cells was associated with a significant decrease of MyD88 protein expression, as well as a reduced production of inflammatory mediator NF\u2010\u03baB 1, TNF\u2010\u03b1 and IL\u20106 in response to BCG infection or LPS stimulation. Further studies using immunoblotting assay against anti\u2010MyD88 antibody and microRNA targeting luciferase reporter assay revealed that miR\u2010149 was able to directly target the 3\u2032\u2010UTR of MyD88 mRNA and post\u2010transcriptionally regulated MyD88 protein expression. These data suggested that miR\u2010149 might be a key player of immune modulator for TLR\/MyD88 signaling pathway in macrophages, which may through a mechanism of negatively regulating MyD88\u2010dependent Toll\u2010like receptors signaling pathway. J. Cell. 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