Neoline protects against doxorubicin-induced heart failure by modulating mitophagy and ferroptosis via the TRIM21/ATAD3A axis

Myocardial pathological changes and fibrosis were observed using H&E and Masson's trichrome staining. Oxidative stress markers (LDH, MDA), ferroptosis-related indicators (Fe²⁺, GSH), and ATP levels in myocardial tissue were determined. Mechanism exploration was conducted through molecular docking simulations and molecular biology validation, with detection of autophagy markers (P62, LC3B), ferroptosis key proteins (GPX4, ACSL4, Nrf2, SLC7A11), as well as TRIM21 and ATAD3A expression levels in myocardial tissue. Neoline (4, 8, and 16 mg/kg) dose-dependently ameliorated DOX-induced cardiac dysfunction, attenuated myocardial pathological damage and fibrosis, and reduced serum levels of BNP, MCP-1, and CRP.

It also reversed oxidative stress and iron accumulation in myocardial tissue. Mechanistically, neoline significantly upregulated LC3B expression and downregulated P62 expression, indicating activation of mitophagy. Concurrently, it upregulated the expression of GPX4, Nrf2, and SLC7A11 while downregulating ACSL4, suggesting inhibition of ferroptosis.

Molecular docking demonstrates that neoline exhibits strong binding affinity to the TRIM21 protein and may function through TRIM21. Further experimental validation confirms that neoline upregulates the expression of TRIM21 while downregulating the expression of ATAD3A. Neoline attenuates DOX-induced heart failure in mice, potentially by activating mitophagy and inhibiting ferroptosis through the regulation of the TRIM21/ATAD3A axis.