Flavonoids have a range of biological activities, including anti-allergic, anti-inflammatory, anti-microbial, and anti-cancer activities, as demonstrated by in vitro studies. In this study, we investigated whether luteolin can be applied to suppression of lipopolysaccharide (LPS)-stimulated inflammatory responses in murine macrophages. Luteolin was found to reduce nitric oxide (NO) production in LPS-stimulated Raw 264.7 cells. In addition, expression of inducible nitric oxide synthetase (iNOS), cyclooxygenase-2 (COX-2), and pro-inflammatory cytokine tumor necrotic factor-α (TNF-α) at the mRNA and protein levels were decreased. These inhibitory effects were found to be caused by the blockade of nuclear factor kappa-light- chain-enhancer of activated B cells (NF-κB) activation and phosphorylation of mitogen-activated protein (MAP) kinase family, extracellular signal-regulated kinases 1/2 (ERK1/2), c-Jun N-terminal kinase (JNK), and p38 MAP kinase. In addition, pre-treatment with luteolin resulted in reduced ganglioside expression levels and inhibited expression of GT1b in Raw 264.7 cells. On the basis of these observations, we suggest that luteolin has potential as an anti-inflammatory drug candidate, and ganglioside GT1b may play a role in the inflammatory process.