Macelignan Improves Functional Recovery After Spinal Cord Injury by Augmenting Autophagy via the AKT‐mTOR‐TFEB Signaling Pathway

ABSTRACT Spinal cord injury (SCI) presents considerable therapeutic challenges due to its complex pathophysiology, and effective treatments are currently lacking. Macelignan (Mace) has shown therapeutic effects in some neurological disorders, but its potential to enhance functional recovery in SCI and the underlying mechanisms are not well understood. This research endeavors to explore the therapeutic value of Mace in SCI and its underlying mechanism of action. A mouse model of SCI was established, and the mice were randomly divided into 13 groups: Sham, Sham + Mace, SCI, SCI + 25 mg/kg Mace, SCI + Mace, SCI + 75 mg/kg Mace, SCI + 100 mg/kg Mace, SCI + 3MA, SCI + Mace/3MA, SCI + Mace/Scramble shRNA, SCI + Mace/TFEB shRNA, SCI + SC79, and SCI + Mace/SC79. Histological examinations were conducted using hematoxylin and eosin (HE), Masson's trichrome, and Nissl staining techniques. Functional recovery post‐injury was evaluated through footprint analysis and the Basso Mouse Scale (BMS). The levels of proteins associated with pyroptosis and autophagy were quantified using qPCR, protein immunoblotting, and immunofluorescence (IF). Network pharmacology techniques were applied to elucidate the signaling pathways modulated by Mace. Mace facilitated functional recovery following SCI by augmenting autophagy and diminishing pyroptosis, with these effects being partially counteracted by 3‐Methyladenine (3MA). It was noted that Mace induced autophagy via inhibition of the AKT‐mTOR signaling pathway, leading to an increase in TFEB expression. As an autophagy activator, Mace induces TFEB‐mediated autophagy and inhibits pyroptosis, which supports functional recovery post‐SCI, indicating its potential clinical relevance.