Aconitine promotes injured peripheral nerve recovery through restraining the activation of inflammasome-mediated cell pyroptosis and pathological inflammation.

Peripheral nerve injury (PNI) represents a common neurological condition with significant social and economic implications. Aconitine, a diterpenoid alkaloid derived from Aconitum species, exhibits potent anti-cancer, anti-viral, anti-inflammatory, analgesic, and immunomodulatory activities against malignancies, rheumatic disorders, arthralgia, and select endocrine pathologies. However, the neuroprotective potential of aconitine in PNI repair remains unclear. Here, we revealed that aconitine treatment at the optimal dose significantly improved SFI values, electrophysiological conduction, axon and myelination regeneration, and cell proliferation and migration. Moreover, aconitine attenuated macrophage polarization towards the M1 phenotype, proinflammatory cytokine secretion, and NLRP3 inflammasome-mediated pyroptosis activation in vivo and in vitro. Mechanistically, RNA sequencing and WB analyses identified the TLR4/MyD88/NF-κB and STING/IRF3 signaling pathways as the potential upstream mediators of anti-inflammatory, anti-inflammasome assembly, and anti-pyroptotic actions of aconitine, which was further verified in vitro experiments. Pharmacological reactivation of either pathway abrogated these therapeutic effects. Thus, aconitine mediates neuroprotection and immunomodulation by polarizing macrophages toward the M2 phenotype and inhibiting NLRP3 inflammasome-driven pyroptosis, mechanisms coordinated through dual blockade of the TLR4/MyD88/NF-κB and STING/IRF3 signaling pathways.