Comparative Metabolomic Profiling of Ocimum Sanctum Leaves Identifies Luteolin As A Neuroprotective Agent Against Neuroinflammation Via NLRP3 Inflammasome Modulation

Abstract

Neuroinflammation is a critical pathological process associated with neuronal dysfunction and progressive neurodegeneration, with NLRP3 inflammasome activation representing an important molecular driver. The present study employed a metabolomics-guided approach to identify bioactive constituents of Ocimum sanctum leaves and evaluate their neuroprotective potential against NLRP3-mediated neuroinflammation. LC-MS/MS-based comparative metabolomic profiling identified multiple flavonoids and phenolic metabolites, among which luteolin exhibited prominent differential enrichment and was confirmed by accurate mass, retention time, and characteristic fragmentation pattern. Targeted quantification demonstrated substantial luteolin abundance in the hydroalcoholic extract. In an LPS-induced BV2 microglial model, luteolin reduced inflammatory activation and suppressed TNF-α, IL-6, IL-1β, and IL-18 production without significant cytotoxicity. Mechanistic evaluation demonstrated attenuation of NF-κB signaling, NLRP3 and ASC expression, and pro-caspase-1 processing, indicating suppression of inflammasome assembly and caspase-1 activation. Luteolin additionally reduced intracellular and mitochondrial ROS, lipid peroxidation, and mitochondrial dysfunction while restoring GSH, SOD, catalase, and mitochondrial membrane potential. In vivo validation further demonstrated reduced microglial activation, inflammatory cytokine production, NLRP3-associated signaling, and neuronal damage, accompanied by improved neurofunctional outcomes. Correlation analysis indicated an association between luteolin enrichment, reduced NLRP3 activation, and improved neuroprotective responses. Collectively, the findings identify luteolin as a prominent neuroprotective constituent of O. sanctum and suggest that coordinated regulation of oxidative stress, NF-κB signaling, and NLRP3 inflammasome activation contributes to its anti-neuroinflammatory activity.