图1 金樱子中含有的7种有效成分的二维结构

Fig.1 2D structure of 7 active ingredients of Rosa laevigata Michx (RLM).

图2 金樱子靶点筛选与特发性肺动脉高压相关性研究

Fig.2 Target screening of RLM for treatment of pulmonary hypertension. A: Venn diagram of the intersecting targets of RLM and idiopathic pulmonary hypertension. B: Construction and screening process of the component-target network. The targets such as SRC and AKT1 show prominent network topological parameters (degree, betweenness centrality), indicating their core roles in regulating smooth muscle cell proliferation.

图3 基于集合靶标构建蛋白质相互作用网络, 并针对核心靶标建立独立网络

Fig.3 Development of the PPI network using the collective targets and a separate PPI network focusing on the core targets.

图5 分子对接分析展示了金樱子核心化合物与PAH中RLM治疗靶点之间的潜在结合模式和相互作用的可靠性

Fig.5 Molecular docking analysis of potential binding modes and reliability of interactions between the core compounds and the targets in RLM treatment of PAH. A: Calculation of the binding energy between 5 targets and 7 active components. The triangles indicate the top 5 binding energies. B: Schematic diagram of the three-dimensional structure of PPARγ-Beta-Sitosterol molecular docking result. C: EGFR-Kaempferol molecular docking result. D: EGFR-Beta-Sitosterol molecular docking result. E: EGFR-Kaempferol molecular docking result. F: SRC-Quercetin molecular docking result.

图 6 金樱子抑制缺氧导致的肺血管平滑肌细胞的增殖

Fig.6 Inhibitory effect of RLM on proliferation of rat pulmonary arterial smooth muscle cells. A: Bar chart showing the effect of different concentrations of RLM on cell viability measured by CCK8 assay. B, C: Western blotting of PCNA in RLM-treated cells. D: Results of CCK-8 assay of RLM-treated rat pulmonary arterial smooth muscle cells under hypoxia condition. E: Quantitative analysis of PCNA expression by immunofluorescence after RLM treatment. F: Immunofluorescence assay of PCNA in RLM-treated cells (scale bar=10 μm). G:Representive image of EdU assay. H: Quantitative analysis bar graph of EdU. All data were analyzed by one-way ANOVA plus Dunnett multiple comparison test. *P<0.05, ***P<0.001.

图 7 RLM在体外抑制低氧诱导的Src 、AKT1磷酸化和降低胞内钙离子浓度

Fig.7 RLM inhibits hypoxia-induced phosphorylation of AKT1 and Src. A: Western blotting of Src and p-Src in RLM-treated cells. B: Quantitative analysis of p-Src/Src expression. C: Western blotting of AKT1 and p-AKT1 in RLM-treated cells. D: Quantitative analysis of p-AKT1/AKT1 expression. E: Calcium ion concentration measurements show a decrease in intracellular calcium levels following RLM treatment. All data were analyzed by one-way ANOVA plus Dunnett multiple comparison test. **P<0.01, ***P<0.001.

图 8 RLM在MCT诱导的肺动脉高压大鼠模型中改善了血流动力学变化

Fig.8 RLM relieves pulmonary hypertension in MCT-induced PAH rat model. A: Schematic diagram and trace of animal experiments. B: Data analysis of RVSP, mRVP and Fulton Index [RV/(LV+S)] in the 5 groups of rats. All data were analyzed by one-way ANOVA plus Dunnett multiple comparison test. *P<0.05, **P<0.01, ***P<0.001.

图 10 RLM抑制了MCT诱导的肺动脉高压大鼠模型中肺血管的增殖

Fig.10 RLM inhibits hypoxia-induced proliferation in MCT-induced PAH rat model. A: Representative images of PCNA and α-SMA immunofluorescence post‐RLM treatment. B: Representative images of HE staining. C. Quantitative analysis of PCNA immunofluorescence post‐RLM treatment.D: Quantitative analysis of α-SMA immunofluorescence post‐RLM treatment. E. Quantitative analysis of HE staining. All data were analyzed by one-way ANOVA plus Dunnett multiple comparison test. *P<0.05, **P<0.01, ***P<0.001.