Platycodin D alleviates cardiac hypertrophy in the treatment of heart failure by inhibiting p53-dependent BCL-2/Bax/caspase-3 signaling pathway

Figures

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  FIGURE 1. Platycodin D (PLD) ameliorates isoprenaline (ISO)-induced heart failure in mice. (A) Schematic of the experimental protocol. (B) Representative cardiac ultrasound images and quantitation results are presented for mice subjected to 2 weeks of PLD pretreatment (at varying doses) followed by 2 weeks of combined ISO co-intervention. Measured parameters include left ventricular ejection fraction (LVEF), left ventricular fractional shortening (LVFS), left ventricular internal diameter at systole (LVIDs), and left ventricular internal diameter at diastole (LVIDd). (C) Representative wheat germ agglutinin (WGA) staining images of cardiac tissue sections reveals group-specific differences in cardiomyocyte hypertrophy. Scale bar = 20 µm. (D) Quantification of cardiomyocyte cross-sectional area of different groups (n = 5). (E) Heart weight-to-tibial length (HW/TL) ratios were calculated to assess cardiac hypertrophy across experimental groups (n = 5). (F) The mRNA expression levels of atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP), and β-myosin heavy chain (β-MHC) in mice cardiac tissues across experimental groups (n = 5). (G) Representative Masson’s trichrome staining…

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  FIGURE 2. Results of network pharmacology analysis. (A) Venn diagram showing the overlapping targets of PLD and cardiac hypertrophy. (B) Protein-protein interaction (PPI) network of the shared targets between PLD and cardiac hypertrophy. (C) PPI network visualized using Cytoscape, where node color intensity correlates with degree centrality (darker nodes indicate higher connectivity). (D) Top 10 significantly enriched Gene Ontology (GO) terms. (E) Top 10 significantly enriched Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways.

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  FIGURE 3. Molecular docking and dynamics simulation analysis of the PLD-p53 interaction. (A) Molecular docking results showing the high-affinity binding pose of PLD with p53. (B) Time-dependent evolution of hydrogen bond interactions between PLD and p53 during the molecular dynamics simulation. (C) Solvent accessible surface area (SASA) of the PLD-p53 complex as a function of simulation time. (D) Radius of gyration (Rg) of the PLD-p53 complex over the simulation trajectory. (E) Root-mean-square deviation (RMSD) of the PLD-p53 complex backbone atoms relative to the initial structure. (F) Root-mean-square fluctuation (RMSF) of PLD-p53 complex residues, reflecting local flexibility.

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  FIGURE 4. Pharmacological inhibition of p53 by PFT-α alleviates cardiac hypertrophy by modulating apoptotic pathways. (A) Inhibition of p53 improves cardiac function, as demonstrated by increased LVEF, LVFS, and decreased LVIDs and LVIDd (n = 5). (B) Representative WGA-stained cardiac sections. Scale bar = 20 µm. (C) Inhibition of p53 reverses ISO-induced increase of cardiomyocyte cross-sectional area (n = 5). (D) Inhibition of p53 reduces the HW/TL ratio (n = 5). (E) Inhibition of p53 suppresses hypertrophic markers, downregulating ISO-induced mRNA expression of ANP, BNP, and β-MHC in cardiac tissues (n = 5). (F) Inhibition of p53 attenuates cardiac fibrosis, as quantified by fibrotic area (n = 4). (G) Inhibition of p53 downregulates fibrotic markers, reducing ISO-induced increases in col1a1 and col3a1 at the mRNA level in cardiac tissues (n = 5). (H) Inhibition of p53 reduces the levels of pro-apoptotic proteins (Bax and caspase-3) while increasing the expression of the anti-apoptotic protein Bcl-2 in cardiac tissues (n = 4). *P < 0.05 vs. Control, #P < 0.05 vs. HF, & P < 0.05 vs. HF + PET-α.

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  FIGURE 5. PLD attenuates ISO-induced cardiachypertrophy by blocking the p53-driven BCL-2/Bax/caspase-3 apoptotic signaling axis. (A) NSC697923 (NSC)-mediated activation of p53 abrogated the cardioprotective effects of PLD, as verified by decreased LVEF, LVFS, and increased LVIDs and LVIDd (n = 5). (B) Representative cardiac sections stained with WGA. Scale bar = 20 µm. (C) Activation of p53 counteracted the inhibitory effect of PLD on cardiomyocyte hypertrophy, as demonstrated by an increase in cardiomyocyte cross-sectional area (n = 5). (D) Activation of p53 results in a significant elevation of the HW/TL ratio (n = 5). (E) Activation of p53 increases the expression of cardiac hypertrophic markers (ANP, BNP, and β-MHC) in cardiac tissues (n = 5). (F) Activation of p53 exacerbated cardiac fibrosis, as indicated by an increase in the fibrotic area (n = 4). (G) Activation of p53 upregulates the expression of fibrosis markers (col1a1 and col3a1) in cardiac tissues (n = 5). (H) Activation of p53 promoted apoptosis by upregulating pro-apoptotic proteins (Bax and caspase-3) while downregulating the anti-apoptotic protein Bcl-2 in cardiac tissues (n = 4). *P < 0.05 vs. Control, #P < 0.05 vs. HF,…