Data Sheet 1_Clemastine alleviates Parkinson’s disease pathology by activating the AMPK/mTOR pathway and enhancing autophagic flux.docx

CLE increased the LC3-II/LC3-I ratio even in the presence of bafilomycin A1, indicating enhanced autophagosome synthesis. Conversely, the AMPK inhibitor dorsomorphin blocked CLE effects, restoring mTOR phosphorylation and α-syn levels. In the chronic MPTP-induced mouse model, CLE administration increased the phosphorylation of AMPK, increased the LC3-II/LC3-I ratio, and decreased mTOR phosphorylation, p62, and α-syn levels.

Immunofluorescence revealed the increased DaNs (tyrosine hydroxylase-positive) and LC3 puncta, alongside reduced the phosphorylated α-syn (p-S129). These cellular and molecular improvements correlated with the alleviation of both motor (rotarod test) and non-motor (fecal pellet output) deficits. Collectively, our findings suggest that CLE alleviates PD pathology by increasing AMPK phosphorylation and decreasing mTOR phosphorylation at Ser2448, suggesting the activation of AMPK pathway and reduced mTOR activity, thereby enhancing the autophagic flux by promoting both the synthesis and degradation of autophagosomes.

Subsequently it facilitates the clearance of pathological α-syn, improves the neuronal survival, and ameliorates the behavioral symptoms. We propose CLE as a promising therapeutic candidate for PD via autophagy modulation.</p>