Integrated in vivo and in silico validation of Corchorus olitorius seed-mediated cardioprotection against doxorubicin-induced cardiotoxicity via network pharmacology, molecular docking, and molecular dynamics

Figures

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  FIGURE 1. Structures of the compounds isolated from the seeds of Corchorus olitorius: (1) β-sitosterol-3-O-β-D-glucoside, (2) stigmasterol, (3) erysimoside, (4) evatromonoside, (5) raffinose, (6) coroloside, (7) corchoroside A, and (8) helveticoside.

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  FIGURE 2. Relative gene expression in the cardiac tissues of different rat groups using qRT-PCR. After normalization to GAPDH, the data reflect the fold changes in expression compared to the control group. The data bars indicate mean ± SD. To ascertain the probability of substantial variations between the study groups, we performed a one-way ANOVA test followed by Bonferroni’s multiple comparisons test, with “*” being p ≤ 0.05 compared to the normal control group and “#” indicating p ≤ 0.05 compared to the doxorubicin-treated group.

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  FIGURE 3. Effects of doxorubicin (DOX) and DOX plus (COR) on cardiac electrophysiological parameters compared to control. (A) Heart rate decreases in DOX-treated groups. (B) PR interval, (C) QRS complex duration, and (D) ST interval are increased in DOX-treated groups. Statistically significant differences versus control are indicated by “a” above the relevant bars.

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  FIGURE 4. Venn diagram illustrates the overlap between heart failure-associated genes and Corchorus olitorius-related genes. A total of 1,476 genes were associated with heart failure, and 313 genes were related to C. olitorius, with 100 genes shared between the two datasets.

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  FIGURE 5. Protein–protein interaction (PPI) network analysis of shared genes. (A) Full PPI network showing all identified proteins as nodes connected by interaction edges. (B) Simplified subnetwork highlighting the top ten hub proteins, color-coded in red, orange, and yellow and outlined with a blue ring, with edges indicating their interactions.

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  FIGURE 6. Molecular docking analysis of compound 6 with the top-ranked target proteins: (A) AKT1, (B) STAT3, (C) IL-1B, and (D) NF-κB. For each target, the left panel shows the three-dimensional binding pose with key interacting amino acid residues and hydrogen bonds, while the right panel shows the corresponding two-dimensional interaction diagram highlighting key residues and interaction sites with colored bonds and labeled atoms.

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  FIGURE 7. Molecular dynamics simulation analysis of compound 6 in the active site of AKT1 over 25 ns. (Top left) Root-mean-square deviation (RMSD) versus time, showing an initial rise followed by stabilization around 0.17–0.20 nm. (Top right) Root-mean-square fluctuation (RMSF) per residue, with a prominent peak near residue 50. (Bottom left) Radius of gyration (Rg) over time, showing a stepwise decline from 2.00 to approximately 1.75 nm. (Bottom right) Number of hydrogen bonds over time, fluctuating between 1 and 9 with a decreasing trend after several peaks.