Compound Centella asiatica formula alleviates Schistosoma japonicum-induced liver fibrosis in mice by inhibiting the inflammation-fibrosis cascade via regulating the TLR4/MyD88 pathway

doi:10.12122/j.issn.1673-4254.2025.06.20

Abstract

Abstract:

Objective To explore the therapeutic mechanism of compound Centella asiatica formula (CCA) for alleviating Schistosoma japonicum (Sj)-induced liver fibrosis in mice. Methods The active components and targets of CCA were identified using the TCMSP database with cross-analysis of Sj-related liver fibrosis targets. A "drug-component-target-pathway-disease" network was constructed using Cytoscape 3.9.1. Functional enrichment analysis (GO/KEGG) was performed using DAVID. Molecular docking study was carried out to validate interactions between the core targets and the key compounds. For experimental validation of the results, 36 mice were divided into control group, Sj-infected model group, and CCA-treated groups. In the latter two groups, liver fibrosis was induced via abdominal infection with Sj cercariae for 8 weeks, followed by 8 weeks of daily treatment with CCA decoction or saline. Hepatic pathology of the mice was assessedwith HE and Masson staining, and hepatic expressions of collagen-I and collagen-III were detected using immunohistochemistry; serum IL-6 and TNF-α levels were determined with ELISA. Hepatic expressions of TLR4 and MyD88 proteins were analyzed with Western blotting. Results We identified a total of 107 bioactive CCA components and 791 targets, including 37 intersection targets linked to Sj-induced fibrosis. The core targets included TNF, TP53, JUN, MMP9, and CXCL8, involving the IL-17 signaling, lipid metabolism, TLR4/MyD88 axis, and cancer pathways. Molecular docking study confirmed strong binding affinity between quercetin (a primary CCA component) and TNF/TP53/JUN/MMP9. In Sj-infected mouse models, CCA treatment significantly attenuated hepatic inflammatory cell infiltration, reduced collagen-I and collagen-III deposition, improved tissue architecture, reduced serum IL-6 and TNF-α levels, and downregulated TLR4 and MyD88 expressions in the liver. Conclusion CCA mitigates Sj-induced liver fibrosis by targeting TNF, TP53, JUN, and MMP9 to modulate the TLR4/MyD88 pathway, thereby suppressing pro-inflammatory cytokine release, inhibiting hepatic stellate cell activation, reducing collagen deposition, and preventing granuloma formation in the liver.

Key words: Schistosoma japonicum-induced hepatic fibrosis, compound Centella asiatica formula, network pharmacology, molecular docking, experimental validation

Liping GUAN, Yan YAN, Xinyi LU, Zhifeng LI, Hui GAO, Dong CAO, Chenxi HOU, Jingyu ZENG, Xinyi LI, Yang ZHAO, Junjie WANG, Huilong FANG. Compound Centella asiatica formula alleviates Schistosoma japonicum-induced liver fibrosis in mice by inhibiting the inflammation-fibrosis cascade via regulating the TLR4/MyD88 pathway[J]. Journal of Southern Medical University, 2025, 45(6): 1307-1316.

Figures/Tables 10

Fig. 1 Venn diagram of the intersection targets between the drug components and disease.

Fig. 2 Drug-component-target-pathway-disease network diagram.

Fig. 3 Analysis of potential target protein interaction.

Fig. 4 GO functional enrichment analysis （A） and KEGG pathway enrichment analysis （B）.

Tab.1 Molecular docking score table (kcal/mol)

Active ingredient	TNF	MMP9	JUN	TP53
quercetin	-5.65	-6.55	-7.06	-5.19
Kaempferol	-5.62	-7.87	-5.39	-5.15
Stigmasterol	-9.52	-8.75	-7.88	-7.02

Fig. 5 Molecular docking simulation of binding between the key components and core targets of compound Centella asiatica formula. A： quercetin-JUN. B： Kaempferol-MMP-9. C： Stigmasterol-TNF. D： Stigmasterol-TP53.

Fig.6 Pathological changes in liver tissue of mouse models with Schistosoma japonicum-induced liver fibrosis (HE and Masson staining, scale bar=200 µm).

Fig.7 Immunohistochemical analysis of expression levels of type I and type III collagen mouse liver tissue （Scale bar=200 µm） and quantitative comparison among the 3 groups. **P<0.01, ***P<0.001 vs Sj, ****P<0.0001 vs control.

Fig.8 ELISA detection of serum levels of IL-6 （A） TNF-α （B） in mouse models with Schistosoma japonicum-induced liver fibrosis. *P<0.05, ***P<0.001.

Fig.9 Effect of compound Centella asiatica formula on hepatic expressions of TLR4 and MyD88 proteins in mouse models of Schistosoma japonicum-induced liver fibrosis detected by Western blotting. **P<0.01, ***P<0.001.

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