Effects of Ferrostatin‐1 on Vocal Folds in Aging Rats

OBJECTIVE: Age-related vocal fold degeneration is characterized by alterations in the lamina propria, extracellular matrix remodeling, and reduced tissue viscoelasticity, contributing to presbyphonia. Ferroptosis, an iron-dependent form of regulated cell death driven by lipid peroxidation, is increasingly implicated in tissue aging. This study investigated the association between ferroptosis and vocal fold aging using ferrostatin-1 (Fer-1) in a rat model. METHODS: Thirty-six male Sprague-Dawley rats were assigned to three groups: Young, Old, and Old-Fer. Fer-1 (2.5 μmol/kg, 20 μL) was injected unilaterally into the left vocal fold at Weeks 0 and 2. At Week 3, vocal folds were harvested for histological, immunohistochemical, and biochemical analyses. Outcome measures included ferroptosis-related markers, lamina propria thickness, extracellular matrix morphology, collagen and hyaluronic acid distribution, and inflammatory cytokines. RESULTS: Compared with the Young group, the Old group showed increased lipid peroxidation, reactive oxygen species, and iron accumulation with reduced glutathione peroxidase 4 activity; these changes were attenuated in the Old-Fer group. The Old group also exhibited increased lamina propria thickness, enhanced collagen deposition, and reduced hyaluronic acid distribution, whereas the Old-Fer group demonstrated reduced collagen accumulation and preserved hyaluronic acid distribution. Transforming growth factor-β and interleukin-1β expression levels were elevated in the Old group and reduced following Fer-1 treatment. CONCLUSION: The results indicate that ferroptosis is involved in vocal fold aging. In addition, Fer-1 effectively suppressed ferroptotic pathways, which may be associated with the attenuation of aging-related extracellular matrix alterations in the vocal folds. LEVEL OF EVIDENCE: NA.