图1 金银花与心力衰竭网络药理学分析

Fig.1 Online pharmacological analysis of Lonicera japonica and heart failure. A. Venn diagram showing the intersection of targets between honeysuckle (JYH) and heart failure. B: Core targets obtained from the visualization analysis of the intersecting genes. C: Combined enrichment plot of biological process (BP), cellular component (CC), and molecular function (MF) obtained by GO enrichment analysis. D: Bubble plot obtained by KEGG pathway enrichment analysis.

图2 金银花水提冻干粉及对照化合物色谱叠加指纹图谱

Fig.2 Superimposed chromatographic fingerprints of Lonicerae Japonicae Flos water extract lyophilized powder and reference compounds.

图3 各组小鼠超声代表图及心功能水平变化

Fig.3 Representative ultrasound images and changes in cardiac function of the mice. A: M-mode echocardiographic images obtained from the parasternal short-axis view in mice across different groups. B: Quantitative statistical analysis of cardiac parameters, including LVEF, LVFS, IVSd, IVSs, LVIDs, LVIDd, LVPWd, and LVPWs (n=6). *P<0.05, **P<0.01, ***P<0.001.

图4 各组小鼠病理切片染色图及心肌损伤和氧化应激分析

Fig.4 Staining images of cardiac tissue sections and oxidative stress assessment. A: HE, Masson, and WGA staining of mouse heart tissues in each group; black arrows indicate inflammatory cell infiltration in HE staining (Scale bar=20 μm). B: Statistical analysis of the proportion of collagen fibers (Masson staining) and cardiomyocyte cross-sectional area (WGA staining) in mouse heart tissues of each group (n=6). C: Analysis of serum CK-MB, α-HBDH, LDH (n=6), MDA, and SOD (n=6) levels in mice of each group. *P<0.05, **P<0.01, ***P<0.001.

图5 各组小鼠心脏组织中炎症相关mRNA和蛋白的表达

Fig.5 Expression of inflammation-related mRNA and protein in cardiac tissues of mice in each group. A: Inflammatory gene expression in mouse heart tissue in each group (n=3). B: Inflammatory protein expression in mouse heart tissue in each group (n=3). *P<0.05, **P<0.01, ***P<0.001.

图6 不同浓度金银花在对OGD后的细胞活力、ROS及凋亡的作用

Fig.6 Effects of honeysuckle at different concentrations on cell viability, ROS production, and apoptosis in OGD-treated cells. A: Cell viability after treatment with different concentrations of Lonicera japonica (n=4). B: ROS staining results of each group after different treatments (n=4) (Scale bar=200 μm). C: Analysis of cell apoptosis in each group (n=4). *P<0.05, **P<0.01, ***P<0.001.

图7 PGC-1α与p-STAT3在小鼠心肌组织及H9C2细胞中的蛋白表达分析

Fig.7 Analysis of protein expression of PGC-1α and p-STAT3 in mouse myocardial tissue and H9C2 cells. A: Protein expression in cardiac tissues of mice in different groups (n=3). B: Protein expression levels in H9C2 cells after different treatments (n=3). *P<0.05, **P<0.01, ***P<0.001.

图8 PGC-1α与p-STAT3在小鼠心肌组织及H9C2细胞中的荧光表达分析

Fig.8 Fluorescence expression analysis of PGC-1α and p-STAT3 in mouse myocardial tissue and H9C2 cells. A, C: Fluorescence expression and quantitative analysis of PGC-1α and p-STAT3 in mouse myocardial tissues (Original magnification: ×40). B,D: Fluorescence expression and quantitative analysis of PGC-1α and p-STAT3 in H9C2 cells (n=3). *P<0.05, **P<0.01, ***P<0.001.

图9 Colivelin逆转金银花的抗氧化应激治疗心衰的作用

Fig.9 Colivelin reverses the antioxidative stress-mediated therapeutic effects of honeysuckle on heart failure. A: ROS production after administration of STAT3 agonist (n=3) (Scale bar=200 μm). B: Expression of p-STAT3 after administration of STAT3 agonist (n=3) (Scale bar=100 μm). **P<0.01, ***P<0.001.

图10 金银花入小鼠心脏组织成分分析及分子对接

Fig.10 Analysis of honeysuckle components entering cardiac tissue and molecular docking results. A: Analysis of components of Lonicera japonica in mouse cardiac tissues in positive and negative ion modes. B: Molecular docking diagrams between main ingredients of three types of Lonicera japonica and STAT3 as well as PGC-1α. C: Effects of three major components on STAT3 and PGC-1α (n=5). **P<0.01, ***P<0.001.