Therapeutic Potential of Vanillylacetone in Ameliorating CCl4-Induced Cardiotoxicity by targeting TNF-α/ NF-κB/TGF-β1 Pathways: Evidence from Biochemical, Histopathological, and Computational Analyses

Figures

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  1. Effects of vanillylacetone on serum LDH and CK-MB against carbon tetrachloride-induced cardiotoxicity. Statistical analysis represented the mean value (n = 8), LDH: (****p < 0.0001 G1vs G2, G4 vs G2; ns p > 0.05 G3 vs G4 & G5 vs G1) and CK-MB: (****p < 0.0001 G1vs G2, G4 vs G2; *p < 0.05 G3 vs G2; ns p > 0.05 G5 vs G1).

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  2. Effects of vanillylacetone on oxidative stress against carbon tetrachloride-induced cardiotoxicity. Statistical analysis represented the mean value (n = 8) and significant value *p < 0.05 G3 vs G2 in GSH; **p < 0.001 G3 vs G2 in MDA, CAT, and SOD; ****p < 0.0001 G2 vs G1 and G4 vs G2; ns p > 0.05 G5 vs G1.

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  3. Effects of vanillylacetone on Inflammatory cytokines (IL-2, IL-6, and TNF-α) against carbon tetrachloride-induced cardiotoxicity. Statistical analysis represented the mean value (n = 8) and significant value *p < 0.05 G3 vs G2 in IL-2; **p < 0.001 G3 vs G2 in TNFa; ****p < 0.0001 G2 vs G1 and G4 vs G2 in IL-2,IL-6, and TNFa; ns p > 0.05 G5 vs G1.

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  4. (A, B) Effects of vanillylacetone on inflammatory cytokines against carbon tetrachloride-induced cardiotoxicity. Statistical analysis representS the mean value (n = 8) and significant value **p < 0.001 G3 vs G2 in NF-kB; ****p < 0.0001 G2 vs G1 and G4 vs G2 in NF-kB and TGF-B1; ns p > 0.05 G5 vs G1 and G3 vs G2 in TGF-B.

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  5. Effects of vanillylacetone on cardiac histology against carbon tetrachloride-induced cardiotoxicity. (G1) normal, (G2) CCl4-induced inflammation and necrosis, (G3 and G4) vanillylacetone treatment and mild inhibition of inflammation and necrosis, and (G5) vanillylacetone without any alteration.

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  6. Illustration of interaction pictures (3D) of vanillylacetone docked with the five target proteins. (a) Interaction with TGF-β1 showing key residues (TYR39, ALA41, ASN103, MET104, and VAL106) involved in hydrogen and alkyl bonding. (b) Interaction with NFK-B1 highlighting main active site residues (ILE224, ARG236, GLY237, and PHE239) forming hydrogen bonds. (c) Interaction with INOS depicting strong binding through residues (TRP194, ALA197, ARG199, CYS200, PHE369, TYR489, and TYR491) via hydrogen bonding, pi-Sigma, and alkyl interactions. (d) Interaction with caspase-3 showing stabilization through residues (THR337, LEU401, and THR467) via hydrogen and alkyl bonds. (e) Interaction with NRF2 demonstrating weak binding through residues (ILE474, LEU476, PRO477, ARG502, and LYS506) with unfavorable interaction at ILE474. These interactions highlight the differential binding affinity of vanillylacetone for the target proteins.

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  7. 2D interaction diagrams of vanillylacetone docked with the five target proteins, highlighting the key binding interactions: (a) vanillylacetone-TGF-β1: interactions include conventional hydrogen bonds with TYR39, ASN103, and MET104 and alkyl interactions with VAL106 and ALA41. An unfavorable Pi-S interaction is observed with MET104. (b) Vanillylacetone-NFK-B1: conventional hydrogen bonds are formed with ARG236, GLY237, and ILE224, alongside pi-alkyl interactions with PHE239. (c) Vanillylacetone-INOS demonstrates strong binding with conventional hydrogen bonds to ARG199, TYR491, and carbon–hydrogen bonds, along with pi-sigma and alkyl interactions involving ALA197 and TRP194. (d) Vanillylacetone-caspase-3: stabilization occurs through hydrogen bonds with THR337 and THR467, with additional pi-alkyl interaction with LEU401. (e) Vanillylacetone-NRF2: weak binding is due to unfavorable acceptor–acceptor interaction with ILE474, supported by conventional hydrogen bonds with ARG502 and alkyl bonds with LEU476 and PRO477.

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  8. Molecular docking interactions of vanillylacetone with various target proteins: (a) TGF-β1, (b) NF-kB1, (c) iNOS, (d) caspase-3, and (e) NRF2. The binding pockets of the target proteins are depicted with a surface representation color-coded to indicate hydrophobic (brown) and hydrophilic (blue) regions. The ligand, vanillylacetone, is shown as a stick model interacting within the binding sites, illustrating key interactions such as hydrogen bonds and hydrophobic contacts.