Vanillin, a kind of phenolic compound, is naturally found in food and beverage and widely used as a flavoring agent. In view of the safety and universality of vanillin, exploring the functions of vanillin on human is of great value. Thus, lipopolysaccharide (LPS)-activated THP-1 cells were selected as the cell model to evaluate the anti-inflammatory effect of vanillin in this study. On the basis of the results, vanillin markedly suppressed the expression of inflammatory cytokines (that is, TNF-α, IL-1β, IL-6, and IL-8), mediators (NO, iNOS, PGE2, and COX-2), and NLRP3 inflammasome (that is, NLRP3, ASC, and caspase-1), blocked the LPS-induced activation of the NF-κB/IκBα/AP-1 signaling pathway, and activated the gene expression of the Nrf2/HO-1 signaling pathway. In addition, it was confirmed that vanillin was unable to react with LPS due to the results of quantification by HS-SPME-GC-MS. Hence, vanillin could effectively attenuate LPS-induced inflammatory response by regulating the expression of intracellular signaling pathways in THP-1 cells. It is a potent anti-inflammatory component found in food and beverage. These findings might contribute to the overall understanding of the potential health benefits of vanillin for food application. PRACTICAL APPLICATION: In this study, the anti-inflammatory effect of vanillin (VA) was evaluated by ELISA, real-time PCR, and western blot in LPS-induced THP-1 cells. The hypothesis that VA could react with LPS was excluded due to the results of quantification by HS-SPME-GC-MS. On the basis of the result, vanillin could effectively attenuate LPS-induced inflammatory response in THP-1 cells and was a potent anti-inflammatory component natural in food and beverage. These findings might contribute to the overall understanding of the potential health benefits of vanillin for food application.