Deciphering the angiogenic potential of Zhishi Xiebai Guizhi decoction in coronary heart disease: an in-depth network pharmacology and experimental investigation

Fig. 1. (A) Venn Zhishi Xiebai Guizhi Decoction and coronary heart disease_Common target of angiogenesis (B) PPI network diagram(C) Degree value top twenty targets (D) Degree value top ten targets.

Fig. 2. (A) "Zhishi Xiebai Guizhi Decoction-Coronary Heart Disease_Angiogenesis Target" network diagram The inverted triangle represents coronary heart disease and angiogenesis, the intersection of the circles represents the treatment target, the diamond represents the active ingredient, and the triangle represents the traditional Chinese medicine Zhishi Xiebai Guizhi Decoction. The connecting lines represent the target and the interaction between the drug and the disease. The more connecting lines, the more targets related to it. (B) Drug-Target-Pathway Diagram In the figure, green represents the target, red represents the disease, orange represents the pathway, purple represents the active ingredient, and the blue triangle represents traditional Chinese medicine.

Fig. 3. (A)Zhishi Xiebai Guizhi Decoction in the treatment of coronary heart disease_GO enrichment analysis of angiogenesis targets (B) Zhishi Xiebai Guizhi Decoction in the treatment of coronary heart disease_KEGG enrichment analysis of angiogenesis targets.

Fig. 4. Molecular Docking Figures Molecular models depicting the binding of EGFR and HIF1A with Luteolin, Naringenin, and Quercetin are presented in both 3D and 2D formats.(A) Thermodynamic binding diagram (B) 3D and 2D interaction diagrams of Luteolin with EGFR (C) 3D and 2D interaction diagrams of Luteolin with HIF1A (D) 3D and 2D interaction diagrams of Naringenin with EGFR (E) 3D and 2D interaction diagrams of Naringenin with HIF1A (F) 3D and 2D interaction diagrams of Quercetin with EGFR (G) 3D and 2D interaction diagrams of Quercetin with HIF1A.

Fig. 5. The administration of ZXGD resulted in increased angiogenesis on HUVECs, contributing to the establishment of the OGD/R model. (A, C) Evaluation of relative cell migration area demonstrated a significant enhancement in HUVEC migration upon treatment with ZXGD (75 μm/L). All experiments were conducted in triplicate, and the data are presented as mean ± S.E.M. (B) Cell viability assays revealed the non-toxic nature of ZXGD on cells, with the highest viability observed at 75 μm/L. (D, E) Representative images of tube formation along with corresponding magnified views were captured for quantitative analysis of total tube length. n = 3 * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Scale bar = 100 μm.

Fig. 6. ZXGD Treatment Enhances Relative Protein Expression (A) The protein expression levels of HIF-1α were assessed through Western blot analysis. This method provided a quantitative measurement of HIF-1α levels in the treated groups. (B) Similarly, the protein expression levels of phosphorylated EGFR (p-EGFR) were evaluated using Western blot analysis, allowing for the assessment of p-EGFR abundance following treatment with ZXGD. (C) The impact of ZXGD on the expression of the HIF-1α signaling protein was observed to be stimulatory. This effect was quantitatively measured and demonstrated an enhancement in HIF-1α protein levels. (D) In a parallel manner, ZXGD treatment also showed a stimulatory effect on the expression of the p-EGFR signaling protein. This indicates an upregulation of p-EGFR protein expression as a response to ZXGD treatment. All experiments were performed in triplicate, at a minimum. Data are presented as mean values ± S.E.M. The results were quantified and presented in bar graph form, showing a significant increase compared to the control group with p < 0.001.

Fig. 7. Enhancement of Dorsal Fin Cell Growth in Zebrafish by ZXGD Treatment (A) Transverse evaluations of Tg(fli1: EGFP) zebrafish embryos conducted 20 h post-fertilization (hpf) under a microscope revealed that ZXGD treatment significantly promotes the growth of dorsal fin vasculature. (B) The relative survival rates of zebrafish embryos subjected to ZXGD treatment (100 μM/L) demonstrated no cytotoxic effects (p < 0.05), indicating the safety of ZXGD at the tested concentration.(C) Following treatment with ZXGD (100 μM/L), a notable increase in the relative length of intersegmental vessels (ISVs) was observed, with statistical significance (p < 0.001), underscoring the efficacy of ZXGD in enhancing vascular development within the dorsal fin region.