Background: Cardiac hypertrophy is a major risk factor for heart failure, and accumulating evidence suggests that ferroptosis—a form of regulated cell death driven by iron—dependent lipid peroxidation-contributes to its progression. However, the regulatory mechanisms linking cardiac hypertrophy and ferroptosis remain incompletely understood. This study, therefore, aimed to investigate the role of anoctamin 1 (ANO1), a calcium-activated chloride channel, in cardiac hypertrophy and ferroptosis.

Methods: A transverse aortic constriction (TAC) mouse model and angiotensin II (AngII)-stimulated H9c2 cardiomyocytes were used to evaluate ANO1 expression. Loss- and gain-of-function experiments were performed in vitro and in vivo. Ferroptosis-related markers, oxidative stress indicators, histological changes, and cardiac hypertrophy parameters were assessed by qPCR, Western blotting, immunohistochemistry, and functional assays.

Results: ANO1 expression was significantly elevated in hypertrophic hearts and AngII-treated cardiomyocytes (p < 0.05). Silencing ANO1 reduced the expression of the ferroptosis-promoting enzyme Acyl-CoA synthetase long-chain family member 4 (ACSL4), restored solute carrier family 7 member 11 (SLC7A11) and glutathione peroxidase 4 (GPX4) levels (p < 0.05), and mitigated oxidative stress and lipid peroxidation, thus leading to improved cell viability and reduced hypertrophy. Conversely, ANO1 overexpression aggravated ferroptosis and oxidative injury (p < 0.05), which could be reversed by the ferroptosis inhibitor ferrostatin-1 (p < 0.05). In vivo, AAV9-mediated ANO1 knockdown attenuated TAC-induced cardiac hypertrophy and ferroptosis (p < 0.05), indicating its pathological role.

Conclusion: ANO1 promotes cardiac hypertrophy by enhancing ferroptosis and oxidative stress. Targeting ANO1 may represent a promising therapeutic strategy for preventing or treating pathological cardiac remodeling.