Vancomycin (VCM), a glycopeptide antibiotic, is commonly applied to infectious diseases caused by Grampositive bacteria, in particular including methicillin-resistant Staphylococcus aureus (MRSA). However, VCM treatment sometimes causes adverse side effects, such as nephropathy and deafness. Although both side effects have been implicated in inflammatory processes, the underlying mechanisms remain unknown. Here, we investigate the cellular responses induced by VCM, especially focusing on the inflammatory responses, and found that VCM promotes the gene expression of NOD-like receptor (NLR) and absent in melanoma 2 (AIM2)-like receptor (ALR) families that mediate release of pro-inflammatory cytokines, interleukin-1β (IL-1β) and IL-18, by forming the multiprotein complexes, called inflammasomes. Thus, our findings suggest that VCM promotes IL-1β- and IL-18-mediated inflammation through the upregulation of the components of inflammasomes, which provides insight into the VCM-induced inflammation-related side effects.