To investigate the role of autologous serum (AS) in alleviating hyperosmolarity-induced corneal epitheliopathy via the ferroptosis pathway.
Methods:
AS was extracted and prepared from patients with Sjögren’s syndrome. An in vitro hyperosmotic dry eye model was established by exposing HCE-T cells to 90 mM NaCl, followed by treatment with AS or ferroptosis inhibitors. Protein expression was analyzed using Western blotting. Intracellular Fe2+ and reactive oxygen species (ROS) levels were determined by FerroOrange and 2’,7’-Dichlorodihydrofluorescein diacetate (H2DCFDA) fluorescent probes, and cellular malondialdehyde and glutathione levels were measured using kits. Transmission electron microscopy was utilized to examine mitochondrial ultrastructure.
Results:
Under hyperosmotic conditions, HCE-T cells exhibited increased lipid peroxidation and iron accumulation, accompanied by significant alterations in the expression of ferroptosis-related markers and characteristic changes in mitochondrial morphology, which indicated the induction of ferroptosis. Among the tested concentrations, 20% and 50% AS demonstrated notable therapeutic effects, significantly enhancing cell viability and migration, reducing ROS and Fe2+ levels, and alleviating mitochondrial damage. Furthermore, AS effectively inhibited ferroptosis through the Xc−/glutathione peroxidase 4 (GPX4) pathway.
Conclusions:
AS reduced lipid peroxidation levels in HCE-T cells under hyperosmolarity and inhibited ferroptosis through the Xc−/GPX4 pathway.
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