Ferulic acid inhibits Tau-441 liquid-liquid phase separation to restore glycolytic metabolism.

Alzheimer's disease (AD) is one of the most common neurodegenerative diseases, and effective clinical treatment currently remains lacking. Ferulic acid (FA) has shown clinical effects in the treatment of AD, but the underlying mechanism of action remains unclear. Here, we showed that FA significantly enhances glycolytic metabolism by inhibiting Tau-441 liquid-liquid phase separation (LLPS). Through molecular docking and solution experiments, we demonstrated that FA binds to Tau-441 via hydrogen bonds, significantly reducing its β-sheet content, thereby effectively preventing the formation of Tau-441 droplets and their transition into solid aggregates. Immunofluorescence staining further confirmed that FA inhibits Tau-441 LLPS formation and reduces the recruitment of hexokinase (HK) during droplet formation. Metabolomic and network analyses revealed that FA significantly alters cellular metabolic profiles, particularly enhancing glycolytic pathway activity. Consistently, FA markedly increased the activities of three key enzymes (HK, PFK, and PK) in the glycolytic pathway, thereby elevating cellular ATP levels and cell viability. These results demonstrate that FA significantly improves glycolytic metabolic efficiency by inhibiting Tau-441 LLPS, thus providing critical experimental and theoretical support for the potential application of FA in treating AD. Overall, this study offers new molecular targets and natural compound intervention strategies for AD treatment.