Acacetin, a flavonoid compound, inhibits inflammation by blocking the MAPK/NF-κB pathways and NLRP3 inflammasome activation. This study investigated the effects of acacetin on NLRP3 inflammasome activation induced by various agonists, including canonical (MSU, ATP, Nigericin) and non-canonical (Pam3CSK4) pathways. The results showed that acacetin significantly reduced the expression of caspase-1, IL-1β, and NLRP3, as well as the release of TNF-α, IL-18, and LDH. Acacetin also suppressed NF-κB p65 phosphorylation and nuclear translocation, inhibited ROS production, and reduced ASC aggregation, thereby suppressing NLRP3 inflammasome activation. Notably, acacetin did not affect K+ and Cl- ion efflux during the activation process. The study demonstrated that acacetin modulates both the priming and assembly processes of the NLRP3 inflammasome by suppressing the NF-κB and MAPK pathways. These findings suggest that acacetin has potential as a therapeutic agent for NLRP3 inflammasome-related diseases.Acacetin, a flavonoid compound, inhibits inflammation by blocking the MAPK/NF-κB pathways and NLRP3 inflammasome activation. This study investigated the effects of acacetin on NLRP3 inflammasome activation induced by various agonists, including canonical (MSU, ATP, Nigericin) and non-canonical (Pam3CSK4) pathways. The results showed that acacetin significantly reduced the expression of caspase-1, IL-1β, and NLRP3, as well as the release of TNF-α, IL-18, and LDH. Acacetin also suppressed NF-κB p65 phosphorylation and nuclear translocation, inhibited ROS production, and reduced ASC aggregation, thereby suppressing NLRP3 inflammasome activation. Notably, acacetin did not affect K+ and Cl- ion efflux during the activation process. The study demonstrated that acacetin modulates both the priming and assembly processes of the NLRP3 inflammasome by suppressing the NF-κB and MAPK pathways. These findings suggest that acacetin has potential as a therapeutic agent for NLRP3 inflammasome-related diseases.