Plantain Extract Inhibits PASMC Proliferation and Improves Pulmonary Artery Remodeling in HPH Rats

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  Figure 1. TOC (Table of Contents) The active ingredient AU in plantain extract can enhance the activity of antioxidant enzymes in the body through its antioxidant effects, reduce oxidative stress levels, inhibit excessive proliferation of pulmonary artery smooth muscle cells (PASMCs), improve pulmonary vascular remodeling, and lower pulmonary artery pressure.

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  Figure 2. Network pharmacological analysis of interaction targets and pathways between PW and HPH. (a) Venn diagram of disease targets and PW action targets. Through screening by GeneCards (Score > 20) and OMIM databases, 5,226 disease‐related targets were obtained. Intersection with 220 targets corresponding to PW active components yielded 179 common targets. (b) Drug‐component‐target‐disease interaction network diagram, comprising 190 nodes (10 active components and 180 common targets), demonstrating the multi‐component and multi‐target characteristics of PW. (c) Protein‐protein interaction (PPI) network of targets, with node color and size set according to degree value, displaying core targets and their interaction relationships. (d) GO functional enrichment analysis bar chart. List 12 enriched entries (p < 0.05) in biological processes (BP), cellular components (CC), and molecular functions (MF) respectively. (e) KEGG pathway enrichment bubble plot. Displays 27 significantly enriched pathways under the condition of p < 0.05, where bubble size represents the number of targets and color indicates p. adjust values.

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  Figure 3. Construction of drug‐compound‐target‐pathway‐disease network and molecular docking. (a) The multi‐layer network diagram illustrates the association of PW active components, target proteins, and related signaling pathways with HPH. The network comprises 189 nodes and 795 edges, encompassing 17 active components, 145 targets, 25 pathways, and disease information. Node shapes distinguish different element types, while color intensity reflects the degree value (Degree) of nodes. (b) Molecular docking binding energy heatmap. The color gradient represents the magnitude of binding free energy (range: −11.3 to +10.1 kcal/mol). A binding energy ≤−5.0 kcal/mol indicates stable binding, while ≤−7.5 kcal/mol suggests high compatibility, providing evidence for strong affinity. (c–f) Molecular docking patterns of PW active ingredients with core targets (AKT1, Bcl2, IL‐1β, IL‐6, TNF, TP53). Protein receptors are shown in purple, active ingredients in green, with key interaction regions (such as hydrogen bonds) highlighted in pink. The results visually demonstrate the specific binding capability between active ingredients and targets at the structural level.

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  Figure 4. Effects of PW extract on pulmonary function, pulmonary artery pressure, pulmonary artery remodeling, and HIF‐1α content in rats with COPD and HPH. (a) FEV0.4/FVC. (b) MMEF. (c) mPAP. (d–f) TGF‐β, ICAM‐1, and MMP‐9 levels in BALF. (g) HE staining of lung tissue. (h–i) WT% and WA%. (j) HIF‐1α content in BALF. (k) Superoxide anion clearance by PW extract components. The results are presented as the mean ± standard deviation (N = 6). * p < 0.05, **p < 0.01, compared with the control group; #p < 0.05, #p < 0.01, compared with the model group.

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  Figure 5. AU protects mitochondria by clearing ROS and reducing oxidative stress. (a–d) The cells in each group were analyzed by flow cytometry. The contents of SOD, CAT, MDA and LDH were detected by ELISA. (e) Fluorescence microscope images of ROS. (f) Fluorescence microscope images of mitochondrial membrane potential.

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  Figure 6. Effects of AU on PASMC migration, proliferation, and apoptosis and Bax and Bcl‐2 expression. (a) The concentration‐response curve of AU in PASMCs was used to determine the IC50. (b) The cell proliferation rate was measured by the CCK‐8 assay. (c) ImageJ software was used to calculate the wound‐healing rate in each group after 72 h of treatment. (d) The apoptosis rate was measured in each group. (e) Images taken 0, 24, 48, and 72 h after the cell monolayer was scratched. (f) The cells in each group were analyzed by flow cytometry. (g) Effects of AU on Bcl‐2 and Bax protein expression. (h) Quantification of intracellular Bcl‐2 levels normalized to tubulin. (i) Quantification of intracellular Bax levels normalized to tubulin. (j) Bax/Bcl‐2 ratio after intracellular quantification.

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  Figure 7. Effects of AU on oxidative stress, pulmonary artery pressure, right ventricular hypertrophy, and pulmonary artery remodeling in the HPH rat model. (a–f) The serum levels of SOD, CAT, MDA, LDH, GSH‐PX, and GR were detected by ELISA. (g) mPAP. (h) Right ventricular myocardial hypertrophy index. (i) HE staining of lung tissue. (j–k) WT% and WA%. (l) Immunofluorescence sections of rat pulmonary arteries from different groups, from top to bottom, show the expression of SOD, CAT, MDA, and LDH; an additional set of images including nuclear proteins and Merge is also provided. (m‐p) Quantitative statistical table of immunofluorescence intensity of SOD, CAT, MDA, and LDH in lung artery sections of rats in different groups.

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  Figure 8. Effects of AU on apoptosis in lung tissue from rats with HPH. (a) Western blotting of rat lung tissue. (b) Quantification of Bcl‐2 levels normalized to tubulin in rat lung tissue. (c) Quantification of Bax levels normalized to tubulin in rat lung tissue. (d) Bax/Bcl‐2 ratio in rat lung tissue. (e) EVG staining of lung tissue. (f) TUNEL staining of lung tissue. (g) Immunohistochemical staining of Bcl‐2 and Bax proteins in lung tissue.