DHDK, a Plant-Derived Natural Small Molecule, Protects Against Doxorubicin-Induced Cardiotoxicity via the PPARG-CPT1B-FAO Axis

Figures

• 

  Figure 1. Integrated network toxicology analysis of DOX targets in cardiotoxicity and chronic disorders. CCK-8 results confirm that DHDK can alleviate cardiomyocyte damage caused by DOX (a), Venn diagram of DOX and potential targets of cardiotoxicity (b), Venn diagrams of DOX and cardiotoxicity, potential targets of cardiac aging (c), and Venn diagrams of potential targets of cardiotoxicity caused by DOX and cardiac aging and lipid metabolism (d). Protein–protein Interaction (PPI) plot (e), Gene Ontology (GO) results (f–h), and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis (i) results of DOX-induced cardiotoxicity and cardiac aging. Data are presented as mean ± SD (n = 3). Statistical significance was determined by Student’s t-test. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, ns indicated no statistical significance.

• 

  Figure 2. In vitro cardioprotective assessment of DHDK followed by network pharmacology analysis. PPI plot (a–c), GO results (d–f), and KEGG enrichment analysis (g) results of DHDK potential target.

• 

  Figure 3. Integrated network toxicology-pharmacology analysis revealing PPARG as a convergent potential target in DOX-induced cardiotoxicity and DHDK-mediated cardioprotection. Venn diagram (a), PPI plot (b–d), GO enrichment analysis (e), and KEGG pathway enrichment analysis (f) of DHDK alleviation of DOX in cardiotoxicity and cardiac aging. Non-covalent molecular docking results (g) and covalent molecular docking results (h) of DHDK and PPARG, a potential alleviation of DOX-induced cardiac injury. The expression level of the potential target protein PPARG was obtained in the Gene Expression Omnibus (GEO) database before and after DOX-induced cardiac injury (i).

• 

  Figure 4. Integrated lipidomics and Hub Gene analysis reveal the potential mechanism of DHDK’s cardioprotection. The separation of each group in lipidomics was good, and the established model was reliable (a–c). Some differential lipid Log2FC values and Variable Importance in the Projection (VIP) values are shown (d). Heatmap visualization of representative lipid levels corrected for protein (e–g). The results of joint analysis of lipidomics, network pharmacology, and network toxicology, among which the arachidonic acid pathway is particularly critical (h). The content of fatty acids closely related to the arachidonic acid pathway in each group was focused on, and the results were visualized in a heatmap (i).

• 

  Figure 5. DHDK exerts cardioprotection by activating PPARG, which enhances fatty acid oxidation (in vitro validation). Cell viability assay (a), β-oxidation-related mRNA expression level (b1–b5), ATP content (c), ROS level (d), mitochondrial membrane potential (e,f) results. Data are presented as mean ± SD (n = 3). Statistical significance was determined by Student’s t-test. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, ns indicated no statistical significance.

• 

  Figure 6. In Vivo Validation of PPARG as a key target for DHDK’s cardioprotection. Experimental dosing scheme for DHDK in animal study (a), HE staining results (b), and TUNEL staining results (c) of myocardial tissue. Scale bar = 40 μm.