To evaluate the anti-inflammation effect of pterostilbene on RAW264.7 macrophages, an inflammation model in vitro was established by lipopolysaccharide (LPS) stimulation. After exposing RAW264.7 cells to pterostilbene and LPS, the mRNA expression of inflammatory cytokines including monocyte chemoattractant protein (MCP)-1, interleukin (IL)-6 and IL-1β, tumor necrosis factor (TNF)-α, and inducible nitric oxide synthase (iNOS) were detected by real-time quantitative polymerase chain reaction (qPCR). Nitric oxide (NO) production in supernatant was analyzed using Griess method. The protein expression of extracellular regulated protein kinases (ERK),c-Jun N-terminal kinase (JNK),p38 mitogen-activated protein kinase (p38 MAPK) and Nuclear factor-κB p65 (NF-κB p65) were detected by Western blot analysis. The results showed that pterostilbene significantly inhibited the gene expression of inflammatory cytokines and the production of NO. Pterostilbene significantly suppressed LPS-induced ERK, p38 and p65 phosphorylation. Data showed that pterostilbene could inhibit the mRNA expression of LPS-induced inflammatory cytokines including MCP-1, IL-6, IL-1β, TNF-α and iNOS, and the release of NO, and its mechanism may be related to blocking the mitogen-activated protein kinase (MAPK) and NF-κB pathway.