Inflammation, while essential for host defense, can become detrimental when dysregulated. MicroRNA-146a (miR-146a) is a well-known anti-inflammatory regulator, primarily targeting the NF-κB pathway. However, its comprehensive mechanisms remain incompletely understood. This study aimed to elucidate the broader impact of miR-146a overexpression on the proteome of lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages using quantitative proteomics. Transfection with a miR-146a mimic resulted in its significant upregulation and the downregulation of known targets, Traf6 and Irak1, at the mRNA level. Global proteomic analysis identified 1232 differentially expressed proteins (DEPs). Pathway enrichment analysis revealed downregulation of proteins associated with IL-6 signaling and NF-κB-related transcription factors (Rel, Nfkb2, Cebpb), alongside decreased expression of pro-inflammatory mediators (Nlrp3, Nos2, Ptgs2). Interestingly, downregulation of Stat2 and interferon-stimulated genes, as well as upregulation of type II interferon signaling components, were also observed. Quantitative RT-PCR and parallel reaction monitoring (PRM) largely validated the proteomic findings for Ptgs2, S100a8, Nos2, Mapkapk2, and Irf3. Functionally, miR-146a overexpression significantly reduced LPS-induced nitric oxide and IL-6 production. These findings demonstrate that miR-146a exerts multifaceted anti-inflammatory effects in macrophages beyond its direct targets in the NF-κB pathway, impacting key inflammatory mediators and potentially modulating interferon signaling, thus providing novel insights into its regulatory mechanisms and therapeutic potential.