MicroRNAs (miRNAs) have been demonstrated to play a pivotal role in the regulation of target gene expression at the post-transcriptional level. In order to better understand the role of miRNA in the immunological regulation of macrophages against Mycobacterium bovis BCG infection, we explored the alteration of immune-related miRNA profile in macrophage RAW264.7 cells in response to BCG infection in this study. Our results demonstrated that miR-142-3p was a potential to negatively regulate the production of pro-inflammatory mediators NF-κB (NF-κB1), TNF-α and IL-6 in the macrophages in part through a mechanism of targeting IRAK-1 gene and post-transcriptionally down-regulating IRAK-1 protein expression.