Luteolin alleviates DSS-induced ulcerative colitis in mice by targeting the NLRP3 inflammasome, as it shows no effect in NLRP3-/- mice. It inhibits NLRP3 activation and IL-1β secretion in macrophages by reducing ROS, mtROS and calcium levels via AMPK binding and signalling. Metabolomic changes suggest lipid metabolism involvement. Luteolin represents a promising NLRP3-targeted therapeutic candidate for UC.