Integrative transcriptomics, machine learning, and molecular docking reveal that nicotine exacerbates ischemic stroke: TLR4-dependent activation of NLRP3/caspase-1/GSDMD-mediated microglial pyroptosis

Objective: This study aimed to clarify the molecular mechanisms through which nicotine (Nic) aggravates ischemic stroke (IS), with an emphasis on inflammation and pyroptosis in Microglia. Methods: An integrative strategy was employed, combining network toxicology for molecular interaction mapping, machine learning for core gene identification, and molecular docking/dynamics for binding validation. These computational predictions were further verified by in vitro and in vivo experiments. Results: Nic was shown to exacerbate IS injury by promoting pyroptosis through activation of the Toll-like receptor 4(TLR4)–myeloid differentiation primary response gene 88(MyD88) and NOD-like receptor family, pyrin domain containing 3(NLRP3) inflammasome pathways, thereby amplifying inflammatory responses. The convergence of computational analyses and experimental findings confirmed the synergistic effect of Nic on vascular injury and neuroinflammation, leading to worsened IS outcomes. Conclusion: Nic accelerates IS progression by modulating pyroptosis and chronic inflammatory signaling. This combined computational–experimental approach provides novel mechanistic insights into Nic-induced stroke pathology and highlights potential molecular targets for therapeutic intervention.