Multitarget mechanisms of the herb pair Achyranthes bidentata and Paeonia lactiflora Pall. in ameliorating hypertensive cardiomyopathy: combining network pharmacology and functional exploration

Figures

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  FIGURE 1. Network pharmacology analysis of the therapeutic targets and pathways of the herb pair Achyranthes bidentata Blume and Paeonia lactiflora Pall. (AB-PL) in the treatment of hypertension. (A) Venn diagram illustrating the overlapping items between hypertension-related genes and potential AB-PL targets. (B) Protein–protein interaction (PPI) network of the common targets. (C) Important core targets identified from the PPI network. (D) Bubble plot showing the top enrichment terms of the Gene Ontology biological processes. (E) Bubble plot displaying the top results of the Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis. (F) Network diagram depicting the component–target–pathway interactions: purple and blue squares represent active compounds; purple and blue triangles denote herb names; red arrows indicate hypertension; red squares signify pathways; green circles represent common target genes.

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  FIGURE 2. Effects of AB-PL on blood pressure, cardiac hypertrophy, and cardiac functions in L-NAME-induced hypertensive mice; (A) systolic blood pressure (SBP); (B) diastolic blood pressure (DBP); (C) mean arterial pressure (MAP); (D) heart weight to bodyweight (HW/BW) ratio; (E) heart weight to tibia length (HW/TL) ratio; (F) atrial natriuretic peptide (ANP) and (G) brain natriuretic peptide (BNP) expression levels; (H) representative echocardiographic images; (I) left ventricular ejection fraction (LVEF); (J) left ventricular fractional shortening (LVFS) (n ≥ 3; # p < 0.05, ## p < 0.01 vs. model group; * p < 0.05, ** p < 0.01 vs. control group).

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  FIGURE 3. Effects of AB-PL on key components of the nitrogen metabolism pathway in L-NAME-induced hypertensive mice: (A) nitric oxide (NO), (B) endothelial nitric oxide synthase (eNOS), and (C) inducible nitric oxide synthase (iNOS) levels (n ≥ 3; # p < 0.05, ## p < 0.01 vs. model group; * p < 0.05, ** p < 0.01 vs. control group).

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  FIGURE 4. Effects of AB-PL on oxidative stress and the Nrf2/HO-1 signaling pathway in the myocardial tissues of hypertensive mice: (A) reactive oxygen species (ROS) level; (B) Nrf2 and (C) HO-1 mRNA expression; (D) superoxide dismutase (SOD) enzyme activity; (E) Keap1, and (F) SOD protein expression. Immunohistochemical staining of (G) Nrf2 and (H) HO-1 (n ≥ 3; # p < 0.05, ## p < 0.01 vs. model group; * p < 0.05, ** p < 0.01 vs. control group).

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  FIGURE 5. Effects of AB-PL on the NLRP3 inflammasome pathway and downstream inflammatory markers in the myocardial tissues of hypertensive mice: mRNA expression levels of (A) NLRP3, (B) ASC, (C) gasdermin D (GSDMD), (D) IL-1β, and (E) IL-18; protein levels of (F) IL-1β and (G) IL-18; immunohistochemical staining of (H) NLRP3 and (I) caspase-1 proteins (n = 3; # p < 0.05, ## p < 0.01 vs. model group; * p < 0.05, ** p < 0.01 vs. control group).

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  FIGURE 6. Metabolic profiling and pathway analyses of serum samples from different treatment groups: (A) principal component analysis (PCA) score plot including quality control (QC) samples; (B) heatmap of representative metabolites among the control, model, and AB-PL groups; (C) key altered metabolic pathways.

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  FIGURE 7. Flowchart of the mechanistic investigation of AB-PL in the treatment of hypertensive cardiomyopathy.