Acacetin, a flavonoid compound, has been shown to inhibit inflammation by blocking the MAPK/NF-κB pathways and NLRP3 inflammasome activation. This study aimed to investigate the effects and mechanisms of acacetin on NLRP3 inflammasome activation induced by various agonists. Mouse bone marrow-derived macrophages (BMDMs) were treated with different concentrations of acacetin and stimulated with Monosodium Urate (MSU), Nigericin, Adenosine Triphosphate (ATP), and Pam3CSK4. Western blot analysis and ELISA were used to measure the expression of pro-caspase-1, pro-Interleukin-1β (Pro-IL-1β), TNF-α, IL-1β, and IL-18. The study found that acacetin inhibited NLRP3 inflammasome activation, reduced the release of inflammatory cytokines, and suppressed the expression of phosphorylated ERK (p-ERK), p-JNK, and p-p38. Acacetin also inhibited NF-κB p65 phosphorylation and nuclear translocation, reduced reactive oxygen species (ROS) production, and prevented ASC oligomerization. Notably, acacetin did not affect the efflux of potassium (K+) and chloride (Cl-) ions during NLRP3 inflammasome activation. These findings suggest that acacetin is a promising candidate for treating NLRP3 inflammasome-related diseases by modulating the NF-κB and MAPK pathways.Acacetin, a flavonoid compound, has been shown to inhibit inflammation by blocking the MAPK/NF-κB pathways and NLRP3 inflammasome activation. This study aimed to investigate the effects and mechanisms of acacetin on NLRP3 inflammasome activation induced by various agonists. Mouse bone marrow-derived macrophages (BMDMs) were treated with different concentrations of acacetin and stimulated with Monosodium Urate (MSU), Nigericin, Adenosine Triphosphate (ATP), and Pam3CSK4. Western blot analysis and ELISA were used to measure the expression of pro-caspase-1, pro-Interleukin-1β (Pro-IL-1β), TNF-α, IL-1β, and IL-18. The study found that acacetin inhibited NLRP3 inflammasome activation, reduced the release of inflammatory cytokines, and suppressed the expression of phosphorylated ERK (p-ERK), p-JNK, and p-p38. Acacetin also inhibited NF-κB p65 phosphorylation and nuclear translocation, reduced reactive oxygen species (ROS) production, and prevented ASC oligomerization. Notably, acacetin did not affect the efflux of potassium (K+) and chloride (Cl-) ions during NLRP3 inflammasome activation. These findings suggest that acacetin is a promising candidate for treating NLRP3 inflammasome-related diseases by modulating the NF-κB and MAPK pathways.