Dual targeting of NCF1 and NLRP3 by roburic acid orchestrates redox homeostasis and inhibits macrophage death in septic lung injury

Sepsis-associated acute lung injury (ALI) is characterized by excessive inflammation and macrophage death, yet precise therapeutic targets remain limited. Single-cell sequencing analysis indicates that, with progression of sepsis-induced lung injury, macrophages exhibit diverse cell-death programs with prominent enrichment of pyroptosis-related signatures. Here, we identify Roburic acid (RBA) as a potent inhibitor of septic ALI and elucidate its mechanism using a nanoparticle delivery system (RBA-NPs). We demonstrate that RBA-NPs significantly attenuate lung injury through a bioactive lipid compound library screening and improve survival in cecal ligation and puncture (CLP)-induced sepsis models. Mechanistically, using chemical proteomics and cellular thermal shift assays, we identify NLRP3 and NCF1 as direct intracellular targets of RBA. Primarily, RBA interacts with the NACHT domain of NLRP3 to directly block inflammasome assembly and pyroptosis. Furthermore, RBA binds to NCF1 to inhibit NADPH oxidase 2 assembly; this restores redox homeostasis, which not only reinforces the suppression of pyroptosis but also confers additional protection by inhibiting lipid peroxidation-mediated ferroptosis. Our study reveals a comprehensive therapeutic strategy where RBA targets the NLRP3 inflammasome while coordinating redox homeostasis via NCF1 to resolve septic lung injury.