Mechanism of S100A9-mediated astrocyte activation via TLR4/NF-κB in Parkinson's disease.

Astrocyte-mediated neuroinflammation plays a key role in Parkinson's disease (PD) progression. The proinflammatory protein S100A9 is linked to various neurodegenerative diseases, but its involvement in astrocyte activation in PD remains unclear. Here, we investigate the role of S100A9 in astrocyte-mediated neuroinflammation in PD. C57BL/6J mice were intraperitoneally injected with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP; 15 mg/kg four times daily) and subsequently treated with Paquinimod, a S100A9 inhibitor (7 mg/kg, once daily for 7 days, totaling 8 doses). We observed an abnormal increase in S100A9 protein expression and a rise in S100A9-positive cells in the striatum of PD mice. Paquinimod treatment significantly improved behavioral deficits (pole test, rotarod test, traction test, and open field tests), prevented the reduction in striatal tyrosine hydroxylase (TH) protein and the loss of dopaminergic neurons (TH) in the substantia nigra (SN) in PD mice. Interestingly, S100A9 was predominantly expressed in astrocytes (GFAPS100A9 cells) rather than in neurons or microglia, and its inhibition significantly reduced astrocyte activation (GFAP cells), reversed A1 astrocyte gene upregulation (H2-D1, C3, Serping1), and increased A2 astrocyte gene expression (Emp1, Ptx3, S100a10). Moreover, S100A9 inhibition also reduced the expression of inflammatory markers (IL-6, IL-1β, TNF-α) and suppressed the TLR4/NF-κB signaling pathway. In vitro, TLR4/NF-κB inhibitors mitigated inflammation and A1/A2 polarization of astrocytic MA cells induced by recombinant S100A9 (rS100A9). These findings suggest that S100A9 mediates astrocyte neuroinflammation and A1/A2 polarization via TLR4/NF-κB signaling, highlighting its potential as a therapeutic target for PD.