Lonicera japonica attenuates oxidative stress and improves post-ischemic heart failure in mice via the STAT3/PGC-1α axis

doi:10.12122/j.issn.1673-4254.2026.07.10

Abstract

Abstract:

Objective To investigate the mechanism by which freeze-dried water extract of Lonicera japonica alleviates oxidative stress and improves post-ischemic heart failure in mice. Methods Bioinformatics analyses were carried out to identify the active constituents in Lonicera japonica, their target genes, and their enrichment pathways. Mouse models of ischemic heart failure established by left anterior descending (LAD) artery ligation were treated with different doses of freeze-dried water extract of Lonicera japonica, and the changes in cardiac function, myocardial pathology and expressions of the STAT3/PGC-1α axis were examined. In cultured H9C2 cells with oxygen-glucose deprivation (OGD), the effects of Lonicera japonica extract on cell viability, reactive oxygen species (ROS) production and apoptosis were evaluated, and its regulatory effects on STAT3 and PGC-1α expression were assessed using Western blotting and immunofluorescence staining; mass spectrometry was employed to analyze the tissue-penetrating components of Lonicera japonica extract, and the binding capacities of its major constituents to STAT3 and PGC-1α were simulated with molecular docking studies. Results In mice with ischemic heart failure, treatment with Lonicera japonica extract significantly improved the cardiac function and alleviated myocardial hypertrophy, inflammatory responses and collagen deposition. The treatment also restored cell viability of H9C2 cells with OGD and reduced cellular ROS production and cell apoptosis. Lonicera japonica extract significantly inhibited STAT3 activation and upregulated PGC-1α expression in H9C2 cells with OGD, and this effect was effectively attenuated by application of the STAT3 agonist Colivelin. Molecular docking results suggested that among the components of Lonicera japonica extract capable of entering the myocardial tissues, lonicerin, chlorogenic acid and 1,5-dicaffeoylquinic acid had strong binding affinity with STAT3 and PGC-1α. Conclusion Lonicera japonica extract alleviates myocardial oxidative stress and improves heart failure in mice via the STAT3/PGC-1α axis, suggesting the value of Lonicera japonica in the treatment of heart failure.

Key words: honeysuckle water extract lyophilisate, STAT3/PGC-1α axis, oxidative stress, heart failure

Miao LI, Guiying YU, Jincheng WANG, Kang XIE, Yulin OUYANG, Neng WANG, Ruijia WEN, Wei WANG, Chun LI. Lonicera japonica attenuates oxidative stress and improves post-ischemic heart failure in mice via the STAT3/PGC-1α axis[J]. Journal of Southern Medical University, 2026, 46(7): 1563-1575.

Figures/Tables 12

Tab.1 Primer sequences for qPCR of IL-6, TNF-α, STAT3, PGC-1α and GAPDH

Primer name	Primer sequence (5'-3')	Product length/bp
IL-6	Forward: GCCTTCTTGGGACTGATGCT	124
	Reverse: AGCCTCCGACTTGTGAAGTG
TNF-α	Forward: ATGGCCTCCCTCTCATCAGT	164
	Reverse: AAGGTACAACCCATCGGCTG
STAT3	Forward: GCAATACCATTGACCTGCCG	115
	Reverse: AACGTGAGCGACTCAAACTG
PGC-1α	Forward: GTCCTTCCTCCATGCCTGAC	128
	Reverse: AGTGCTAAGACCGCTGCATT
GAPDH	Forward: TGATGGGTGTGAACCACGAG	152
	Reverse: AGTGATGGCATGGACTGTGG

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.

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

Tab.2 Results of linearity investigation and content calculation of each compound

Compound	Regression equation	R²	Peak area of sample	X value (μg)	Concentration (mg/mL)	Content (%)
Chlorogenic acid	Y=1×10⁶X-92498	0.9998	1528159	1.6207	0.1621	4.0516
Isochlorogenic acid A	Y=2×10⁶X-10858	0.9998	289912	0.1504	0.0150	0.3759
Isochlorogenic acid C	Y=4×10⁶X-8386.9	0.9999	419779	0.1070	0.0107	0.2676
Luteoloside	Y=3×10⁶X-9093.8	0.9998	145547	0.0515	0.0052	0.1289
Swertiamarin	Y=2×10⁶X-22051	0.9998	222565	0.1223	0.0122	0.30577

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.

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.

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.

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.

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.

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.

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.

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.

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