Protective effect of Lonicerae japonicae flos extract against doxorubicin-induced liver injury in mice

doi:10.12122/j.issn.1673-4254.2024.08.16

Abstract

Abstract:

Objective To explore the mechanism underlying the protective effect of Lonicerae japonicae flos (LJF) extract against doxorubicin (DOX)-induced liver injury in mice. Methods Network pharmacology methods were used to obtain the intersection genes between LJF targets and disease targets, based on which the protein-protein interaction (PPI) network was constructed using STRING database for screening the core targets using Cytoscape software. DAVID database was used for bioinformatics analysis, and the core components and core targets were verified using molecular docking study. In a mouse model of DOX-induced liver injury, the effect of LJF extract on liver pathologies, serum levels of ALT and AST, and hepatic expressions of HYP, ROS, TNF-α, IL-6, COL-IV and P53 proteins were evaluated using HE and Masson staining, ELISA, and Western blotting. Results We identified 12 core targets from 43 intersection genes involving cancer pathway, IL-17 signaling pathway, and TNF signaling pathways. Molecular docking study suggested that 10 core components of LJF could bind to different core targets. The mice with DOX-induced liver injury showed elevated serum AST and ALT levels with obvious liver injury and fibrosis, increased ROS content, and enhanced expressions of TNF-α, IL-6, HYP, COL-IV and P53 proteins in the liver tissue. All these changes in the mouse models were significantly alleviated by treatment with LJF extract, suggesting obviously lowered levels of oxidative stress, inflammation and fibrosis in the liver tissues. Conclusion LJF extract is capable of alleviating DOX-induced liver injury in mice by downregulating Trp53, TNF and IL-6 to reduce liver oxidative stress, inflammation and fibrosis.

Key words: network pharmacology, Lonicerae japonicae flos, doxorubicin, liver damage, oxidative stress, liver fibrosis, inflammation

Yuming ZHANG, Shicheng XIA, Linlin ZHANG, Mengxi CHEN, Xiaojing LIU, Qin GAO, Hongwei YE. Protective effect of Lonicerae japonicae flos extract against doxorubicin-induced liver injury in mice[J]. Journal of Southern Medical University, 2024, 44(8): 1571-1581.

Figures/Tables 12

Fig.1 Venn diagram of Lonicerae japonicae flos, Adriamycin-induced liver injury and hepatic fibrosis-related targets.

Fig.2 PPI network diagram (A) and core targets network diagram (B).

Fig.3 GO (A) and KEGG (B) enrichment analysis.

Tab.1 Core components of Lonicerae japonicae flos

Mol ID	Molecule	OB	DL
MOL000006	luteolin	36.16	0.25
MOL000098	quercetin	46.43	0.28
MOL000422	kaempferol	41.88	0.24
MOL002914	Eriodyctiol (flavanone)	41.35	0.24
MOL003006	(-)-(3R,8S,9R,9aS,10aS)-9-ethenyl-8-(beta-D-glucopyranosyloxy)-2,3,9,9a,10,10a-hexahydro-5-oxo-5H,8H-pyrano[4,3-d]oxazolo[3,2-a]pyridine-3-carboxylic acid_qt	87.47	0.23
MOL003014	secologanic dibutylacetal_qt	53.65	0.29
MOL003044	Chryseriol	35.85	0.27
MOL003095	5-hydroxy-7-methoxy-2-(3,4,5-trimethoxyphenyl)chromone	51.96	0.41
MOL003111	Centauroside_qt	55.79	0.5
MOL003117	Ioniceracetalides B_qt	61.19	0.19

Tab.2 Binding energy of the core targets and core ingredients (kcal/mol)

Mol ID	TrP53 Binding energy	TNF Binding energy	IL-6 Binding energy
MOL000006	-4.49	-5.8	-3.93
MOL000098	-3.93	-5.6	-4.24
MOL000422	-4.85	-5.62	-4.59
MOL002914	-5.61	-5.09	-4.75
MOL003006	-4.7	-6.97	-5.31
MOL003014	-2.53	-2.96	-0.83
MOL003044	-5.02	-5.99	-5.1
MOL003095	-4.7	-4.9	-3.59
MOL003111	-2.85	-4.11	-3.23
MOL003117	-4.94	-4.79	-4.24

Fig.4 Visualization of molecular docking results. A: Potential interaction between MOL002914 and Trp53. B: Potential interaction between MOL003044 and Trp53. C: Potential interaction between MOL003044 and TNF. D: Potential interaction between MOL003006 and TNF. E: Potential interaction between MOL003006 and IL-6. F: Potential interaction between MOL003044 and IL-6.

Fig.5 Serum levels of AST (A) and ALT (B) in mice in each group (Mean±SD, n=6). ***P<0.001 vs Sham; ###P<0.001 vs DOX; $P<0.05, $$P<0.01 vs DOX+LJF (0.2 g/kg); &&P<0.01 vs DOX+LJF (0.4 g/kg).

Fig.6 HE staining of mouse liver tissues in each group (Original magnification: ×40). Black arrows indicate erythrocyte exudates, red arrows indicate hepatocyte ballooning, and blue arrows indicate inflammatory cell infiltration.

Fig.7 Masson staining of mouse liver tissue in each group (×120). The black arrows indicate fibrotic hyperplasia and the red arrows indicate hepatocyte balloons.

Fig.9 TNF-α (A) and IL-6 (B) levels in mouse liver tissues in each group (Mean±SD, n=6). ***P<0.001 vs Sham; ###P<0.001 vs DOX; $$P<0.01, $$$P<0.001 vs DOX+LJF (0.2 g/kg) ; &&P<0.01 vs DOX+LJF (0.4 g/kg).

Fig.10 HYP (A) and COL-IV (B) levels in mouse liver tissues in each group (Mean±SD, n=6). ***P<0.001 vs Sham; ###P<0.001 vs DOX; $P<0.05, $$P<0.01, $$$P<0.001 vs DOX+LJF (0.2 g/kg) ; &P<0.05, &&P<0.01 vs DOX+LJF (0.4 g/kg).

Fig.11 Protein expression level of P53 in mouse liver tissue in each group (Mean±SD, n=6). A: P53 protein expression detected by Western blotting in each group. B: Quantitative data of P53 expression. ***P<0.001 vs Sham; ###P<0.001, ##P<0.01, #P<0.05 vs DOX; &P<0.05 vs DOX+LJF (0.2 g/kg).

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