Protective effects of Aronia melanocarpa seed oil against doxorubicin-induced cardiotoxicity in H9c2 cells: insights from apoptosis-related mRNA expression and molecular docking analyses

Purpose: This study aimed to investigate the protective effects of Aronia melanocarpa (chokeberry) seed oil (ASO) against doxorubicin (DOX)-induced cardiotoxicity in H9c2 cells by evaluating apoptosis-related mRNA expression and assessing the potential interactions of ASO-derived fatty acids with apoptosis-related proteins using molecular docking.Materials and Methods: ASO cytotoxicity was assessed using the MTT assay, and concentrations of 10, 25, and 50 µg/mL were selected for further experiments based on the IC₅₀ value (377 µg/mL). H9c2 cells were pretreated with ASO for 30 min before exposure to 5 µM DOX for 24 h. Cell viability was assessed using the MTT assay, and the mRNA expression levels of caspase-3, Bax, and Bcl-2 were determined using RT-qPCR. Molecular docking analyses were performed to evaluate the binding affinities of ASO-derived fatty acids toward apoptosis-related proteins.Results: DOX increased caspase-3 and Bax mRNA expression (~3.31-fold and ~1.62-fold vs. control, respectively) while decreasing Bcl-2 expression (~0.42-fold vs. control). ASO pretreatment dose-dependently attenuated these alterations, with the strongest effects observed at 50 µg/mL, reducing caspase-3 and Bax mRNA expression (~0.36-fold and ~0.64-fold vs. DOX, respectively) while increasing Bcl-2 expression (~2.14-fold vs. DOX). In addition, ASO increased the Bcl-2/Bax ratio compared with the DOX-treated group (~1.25-, ~2.62-, and ~3.31-fold vs. DOX at 10, 25, and 50 µg/mL, respectively). Molecular docking demonstrated potential interactions between ASO-derived fatty acids and apoptosis-related proteins, with binding energies ranging from −4.8 to −5.8 kcal/mol.Conclusion: ASO attenuated DOX-induced apoptotic responses by modulating apoptosis-related mRNA expression, supporting its potential as a natural cardioprotective agent. Further in vivo studies are warranted to confirm these findings and clarify the underlying cardioprotective mechanisms.