Therapeutic mechanism of hederagenin, an active component in Guizhi Fuling Pellets, against cervical cancer in nude mice

doi:10.12122/j.issn.1673-4254.2025.07.08

Abstract

Abstract:

Objective To explore the therapeutic mechanism of Guizhi Fuling (GZFL) Pellets against cervical cancer. Methods Publicly available databases were used to identify the targets of GZFL Pellets and cervical cancer to construct the protein-protein interaction (PPI) network, followed by GO biological process and KEGG pathway enrichment analysis of the hub genes. The "Traditional Chinese Medicine-Active Ingredients-Targets-Pathways" network for GZFL Pellets in cervical cancer treatment was generated using Cytoscape v10.0.0, and molecular docking of the drug and potential targets was performed to predict the specific targets of active components in Guizhi Fuling Pellets. The inhibitory effects of hederagenin, an active ingredient in GZFL Pellets, was tested in cultured cervical cancer cells and in nude mice bearing cervical cancer xenografts. Results GZFL Pellets contain 338 active components targeting 247 action sites. A total of 10127 cervical cancer-related targets were obtained, and among them 195 were identified as potential therapeutic targets of GZFL Pellets for cervical cancer treatment, including the key targets of GABRA1, PTK2, JAK2, HTR3A, GSR, and IL-17. Molecular docking study showed low binding energies of the active components such as hederagenin, campesterol, and stigmasterol for protein-molecule interaction. GO enrichment analysis suggested that GZFL Pellets inhibited cervical cancer primarily by regulating responses to steroid hormones, oxidative stress, and lipopolysaccharides. Among the active components of GZFL Pellets, hederagenin was found to inhibit cervical cancer cells in vitro and significantly reduced STAT3 phosphorylation level in the cancer cells. In nude mice bearing cervical cancer xenografts, hederagenin effectively inhibited tumor growth rate without causing obvious adverse effects. Conclusion GZFL Pellets inhibit cervical cancer cell growth through its multiple active components that target different pathways. Among these components, hederagenin inhibits tumor cell growth possibly by directly binding to JAK2 protein to inhibit STAT3 phosphorylation.

Key words: network pharmacology, Guizhi Fuling Pellets, cervical cancer, HPV infection, hederagenin

Yinfu ZHU, Yiran LI, Yi WANG, Yinger HUANG, Kunxiang GONG, Wenbo HAO, Lingling SUN. Therapeutic mechanism of hederagenin, an active component in Guizhi Fuling Pellets, against cervical cancer in nude mice[J]. Journal of Southern Medical University, 2025, 45(7): 1423-1433.

Figures/Tables 9

Fig.1 Construction of the "Traditional Chinese Medicine-Active Ingredients-Targets" Network. A:Venn diagram of the drug targets and cervical cancer targets. B: Traditional Chinese medicine-active ingredient-target network of Guizhi Fuling Pellets against cervical cancer.

Tab.1 Topology parameters of the herbs and compounds in the network

Name	Betweenness centrality	Closeness centrality	Degree	Eccentricity	Neighborhood connectivity	Number of undirected edges	Type
Quercetin	0.12874832	0.465960666	44	4	32.38636364	44	ingredient
Beta-sitosterol	0.042529052	0.447024673	26	4	60.03846154	26	ingredient
Kaempferol	0.054556646	0.450951684	31	4	50.90322581	31	ingredient
(+)-Catechin	0.01968019	0.426002766	15	5	94.73333333	15	ingredient
Hederagenin	0.108300219	0.443165468	17	5	59.58823529	17	ingredient
Stigmasterol	0.011819554	0.421340629	16	5	62.625	16	ingredient
Sitosterol	0.011819554	0.421340629	16	5	62.625	16	ingredient
Luteolin	0.041269784	0.42837274	22	4	51.40909091	22	ingredient
Baicalein	0.022662695	0.422496571	18	4	76.83333333	18	ingredient
Campesterol	0.003171002	0.417910448	10	5	112.4	10	ingredient

Fig.2 GO and KEGG enrichment analysis of the intersection targets. A: PPI network constructed and visualized using Cytoscape v10.0.0. Nodes are colored based on the degrees. The degree values range from large to small, following a continuous mapping from yellow to pink. B: Top 10 items for the biological process (BP), cellular component (CC), and molecular function (MF) enrichment analysis in the GO enrichment analysis. The orange section represents the biological processes. The green part represents the cellular component. The blue part represents the molecular function. C: Top 10 items of the KEGG pathway enrichment analysis.

Fig.3 Molecular docking results of the target proteins and the active molecules. A: Heatmap of the molecular docking fraction showing the binding energy of target proteins and active compounds (kcal/mol). B: Molecular docking model for JAK 2 and hederagenin. C: Molecular docking model for GSR and campesterol. D: Molecular docking model for HTR 3 A and beta-sitosterol.

Tab.2 Binding energy between the predicted active components and targets

Sample	GABRA1	PTK2	JAK2	HTR3A	GSR	IL-17
(+)-catechin	-8.1	-6.7	-8.7	-8.6	-8.7	-9.5
Baicalein	-8.1	-6.9	-9.4	-8.8	-8.7	-9.2
Beta-sitosterol	-4.3	-8.2	-9.6	-10.8	-10.7	-10.4
Campesterol	-4.6	-8	-9.4	-11.3	-10.5	-9.2
Hederagenin	-8.6	-6.9	-10.7	-9.4	-11.1	-10.2
Kaempferol	-7.7	-7	-8.8	-8.5	-8.9	-8.8
Luteolin	-8.5	-8.5	-9	-9	-9	-9.1
Sitosterol	-4.3	-7.6	-9.4	-10.2	-10.8	-9.4
Quercetin	-8.1	-8.9	-9.6	-9.2	-9.1	-8.8
Stigmasterol	-4.7	-8.6	-10.0	-10.8	-11	-9.3
Isocorypalmine	-8.0	-8.2	-8.2	-8.9	-9.7	-8.7

Tab. 3 Active curpockets of the amino acids

	JAK2-hederagenin	GSR-campesterol	HTR3A-beta-sitosterol
Chain A	LYS607 HIS608 GLU652 LYS655 GLN656 TRP659 HIS662 GLU666 PRO692 PHE694 PHE798 ARG799 ALA800 ILE802 ARG803 ASN806	LYS67 TRP70 ASN71 VAL74 PHE78 PRO376 PRO405 MET406 TYR407 LEU438 GLY439 ASP441 GLU442 PRO468 SER470	PHE244 VAL247 SER248 LEU249 LEU251 PRO252 PHE255 LEU256 THR279 LEU280 LEU282 GLY283 TYR284 VAL286 PHE287 ILE290 VAL291 ASP293 THR294
Chain B	ASP768 ARG769 HIS770 GLN771 LEU772 ASP789 TYR790 GLU791 HIS794		LEU281 SER285 LEU288 VAL291 SER292 LEU295 PRO296 ALA297 THR298 ALA299 GLY301 THR302 PRO303 GLY306 VAL307 PHE309 VAL310 MET313 ALA314 VAL317

Fig. 4 Hederagenin inhibits growth of cervical cancer cells in vitro. A: IC50 curve of hederagenin against U14 cells. B, C: Viability of U14 cells treated with high-concentration (B) and low-concentration (C) hederagenin. D, E: Western blotting and quantitative analysis of STAT3 phosphorylation inhibited by hederagenin treatment for 72 h. *P<0.05, ***P<0.001.

Fig.5 Hederagenin inhibits growth of cervical cancer xenografts in nude mice. A: Gross observation of the dissected tumors. B: Tumor growth curve during hederagenin treatment. C: HE staining and Ki-67 immunohistochemistry of the tumor sections. D: Weight of the tumors. E: Body weight curve of the nude mice during hederagenin treatment. *P<0.05.

Fig. 6 HE staining results of the liver, spleen and kidney sections from the tumor-bearing mice.

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