Baicalin and kaempferol alleviates cuprizone-induced demyelination and microglial activation by inhibiting the STAT3 and NF-κB signaling pathways

Microglia-mediated neuroinflammation plays a critical role in myelin loss in multiple sclerosis (MS). Baicalin (BAI) and kaempferol (KAE) are known for their potent anti-inflammatory properties; however, their potential to alleviate demyelination and treat MS has rarely been rarely investigated.

Demyelination was induced in mice using cuprizone (CPZ), and behavioral assessments were conducted to evaluate motor function recovery following treatment with BAI and KAE. Myelin loss was assessed using transmission electron microscopy. Immunofluorescence and Western blot analyses were employed to assess microglial activation and neuroinflammatory responses. Moreover, network pharmacology, molecular docking, and Drug Affinity Responsive Target Stability (DARTS) were utilized to identify molecular targets involved in the therapeutic effects of BAI and KAE.

Both BAI and KAE significantly improved motor function and mitigated myelin loss in CPZ-treated mice. These compounds effectively inhibited CPZ-induced microglial activation and attenuated secretion of pro-inflammatory cytokines. Network pharmacology identified the STAT3 and NF-κB as potential targets of BAI and KAE. In vivo and in vitro validation further confirmed that both compounds significantly suppressed the phosphorylation of STAT3 and NF-κB p65 proteins.

Our findings suggest that BAI and KAE alleviate CPZ-induced demyelination by inhibiting STAT3 and NF-κB signaling pathways, thereby reduced microglial activation and alleviation of neuroinflammation.