Rosmarinic Acid Alleviates Doxorubicin-Induced Cellular Senescence and Cardiotoxicity by Targeting the 14-3-3/Foxo1 Signaling Axis

Abstract

Background: Doxorubicin (DOX)-induced cardiotoxicity remains a major clinical challenge in cancer chemotherapy, with cardiomyocyte senescence as an emerging pathogenic mechanism. Rosmarinic Acid (RA), a polyphenolic compound with demonstrated anti-aging properties; however, its precise molecular mechanisms in targeting cellular senescence remain to be fully elucidated.

Purpose: This research aimed to explore how RA affects cardiomyocyte senescence and its treatment efficacy in DOX-induced cardiotoxicity.

Study design and methods: In the DOX-induced senescent HL-1 cardiomyocyte model, the anti-senescence effects of RA were assessed by CCK8, Senescence-Associated β-galactosidase (SA-β-gal) staining, RT-qPCR and ELISA. The molecular targets underlying RA's anti-senescence activity were examined by Activity-Based Protein Profiling (ABPP), Cellular Thermal Shift Assay (CETSA), Microscale Thermophoresis (MST), western blot and molecular docking. The therapeutic effect of RA was investigated in the DOX-induced cardiotoxicity mouse model by evaluating survival rate, echocardiographic, heart failure biomarkers, senescence symptoms, histopathology and immunofluorescence analysis.

Results: Through cellular screening of natural polyphenolic compounds, we identified and confirmed RA as an effective inhibitor of DOX-induced cardiomyocyte senescence by decreased SA-β-gal activity, downregulating p16/p21 and attenuating SASP secretion (Il-6, Il-1β, Tnf-α). Mechanistically, RA upregulated 14-3-3θ expression, enhanced Foxo1 phosphorylation, and inhibited its nuclear translocation, thereby attenuating the activation of senescence-associated signaling pathways. In a murine model of DOX-induced cardiotoxicity, RA administration markedly suppressed cardiomyocyte senescence, improved survival rates, maintained cardiac functional parameters, and attenuated key pathological features including myocardial atrophy, fibrotic remodeling, and mitochondrial impairment.

Conclusion: Our findings established RA as a promising therapeutic agent for preventing DOX-induced cardiotoxicity through the modulation of the 14-3-3/Foxo1 signaling axis, offering a novel pharmacological approach to enhance the safety profile of current cancer chemotherapy.