In Silico, In Vitro, and In Vivo Evaluation of <i>Woodfordia Fruticosa</i> for Cardio Protection against Doxorubicin‐Induced Cardiomyopathy

This study aimed to evaluate the therapeutic potential of Woodfordia fruticosa, known for its cardioprotective properties, using in silico, in vitro, and in vivo methods. Fourier-transform infrared spectroscopy (FTIR) and gas chromatography-mass spectrometry (GC-MS) analyses were used to identify and characterize the bioactive compounds in W. fruticosa. Molecular docking with dock score assessed the binding affinities of hecogenin, β-sitosterol, and pulmatin. Antioxidant potential was evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP) assays. In vivo experiments involved inducing cardiomyopathy in Wistar albino rats with doxorubicin (16 mg/kg, ip) and treating them with W. fruticosa flower extract at doses of 100, 200, and 400 mg/kg for 16 days. Molecular docking studies revealed that hecogenin, β-sitosterol, and pulmatin exhibited strong binding affinities against human protein kinase ZAK, with Dock scores of -12.07, -11.42, and -11.04, respectively, surpassing the standard drug ramipril (-9.32). FTIR and GC-MS confirmed the presence of key phytoconstituents, including β-sitosterol, hecogenin, ellagic acid, kaempferol, and 2-hydroxy-1,4-naphthoquinone (Lawsone). In vitro antioxidant assays demonstrated significant free radical scavenging activity comparable to ascorbic acid. In vivo studies in Wistar albino rats showed that W. fruticosa flower extract significantly (p < 0.001) improved electrocardiogram (ECG) parameters, and significantly (p < 0.001) reduced cardiac biomarkers, including creatine kinase myocardial band (CK-MB), lactate dehydrogenase (LDH), and cardiac troponin I, and mitigated histopathological damage in a dose-dependent manner caused by doxorubicin. The 400 mg/kg dose exhibited the most pronounced cardioprotective effects, normalizing ECG patterns, decreasing myocardial injury, and reducing fibrosis. W. fruticosa flower extract exhibited significant cardioprotective potential via the human protein kinase ZAK pathway against doxorubicin-induced cardiomyopathy through in silico, in vitro, and in vivo evaluations. These findings suggest W. fruticosa as a promising natural agent for cardiomyopathy management, warranting further studies to elucidate its mechanisms, optimize dosing, and validate long-term safety and efficacy.