Mechanosensitive ion channel TRPV4 inhibition alleviates glaucomatous inflammation by regulating microglial ferroptosis.

Glaucoma is a neurodegenerative disease characterized by the progressive and irreversible loss of retinal ganglion cells (RGCs). Elevated intraocular pressure (IOP) is a key factor in initiating pathological processes in acute glaucoma. This study aims to investigate the role of TRPV4 in mediating retinal inflammatory damage following acute glaucomatous injury. Using an in vitro elevated hydrostatic pressure (EHP) model established in BV2 microglia and an in vivo acute ocular hypertension (AOH) model in mice, we assessed microglial migration and polarization (M1/M2), lipid peroxidation, mitochondrial morphology, ferroptosis-related protein expression, and retinal ganglion cell survival. The function of TRPV4 was validated using the specific inhibitor HC-067047 and TRPV4 gene knockdown, and the p53/SLC7A11/GPX4 ferroptosis axis was investigated using the ferroptosis inhibitor Ferrostatin-1 (Fer-1), the p53 agonist Nutlin-3, and p53 gene knockdown. Results demonstrated that injury upregulates and activates TRPV4, inducing calcium influx in microglia. TRPV4 activation triggered ferroptosis by upregulating p53 and downregulating SLC7A11 and GPX4, thereby promoting pro-inflammatory M1 microglial polarization and exacerbating ganglion cell injury. Notably, pharmacological inhibition of TRPV4 reversed these responses, shifting microglia toward an anti-inflammatory M2 phenotype and mitigating ganglion cell loss. These findings demonstrate that TRPV4 activation drives ferroptosis via the p53/SLC7A11/GPX4 axis, thereby promoting microglial-mediated neuroinflammation, providing a new theoretical perspective for understanding the pathogenesis of neuroinflamation in glaucoma.