Weifuchun (胃复春 ) exerts therapeutic effects on gastric fundic gland polyps by promoting ferroptosis

doi:10.19852/j.cnki.jtcm.20250515.001

Abstract

Abstract:

OBJECTIVE: To investigate the therapeutic effects of Chinese medicine Weifuchun (WFC, 胃复春) on gastric fundic gland polyps (FGPs).
METHODS: FGPs organoids were constructed with patients-derived samples. The morphology and size of FGPs organoids were detected using bright-field imaging. Effective components and corresponding potential targets of WFC were screened using multiple open-source databases and research on Traditional Chinese Medicine or compound formulas. Core genes were identified through protein-protein interaction networks. Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) enrichment analyses of the core genes were conducted. The interactions between main components and core targets were analyzed through the FerrDb database. The expressions of core targets were detected by quantitative real-time polymerase chain reaction (qRT-PCR).
RESULTS: After WFC treatment, the number and size of FGPs organoids were significantly reduced. Twenty nine active drug components and 162 candidate targets of WFC for treating FGPs were identified, including 37 targets related to ferroptosis. Quercetin, Glaucocalyxin B, Melissoidesin U, Melissoidesin O, Hesperetin, Glaucocalyxin A, Angustifolin, Melissoidesin M, Di-n-octyl phthalate, and beta-sitosterol were identified as the main active compounds. SRC proto-oncogene, non-receptor tyrosine kinase, signal transducer and activator of transcription 3, phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha, phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit beta, phosphoinositide-3-kinase regulatory subunit 1, and AKT serine/threonine kinase 1 were identified as the primary targets. KEGG pathways related to carcinogenesis, cell proliferation and metabolism, and oxidative stress. WFC promoted FGPs organoids’ death and could be reversed by ferroptosis inhibitor of Erastin. The qRT-PCR results showed that WFC treatment could regulate the mRNA expression levels of solute carrier family 7 member 11, acyl-CoA synthetase long chain family member 4, and arachidonate 15-lipoxygenase, type B.
CONCLUSION: WFC may exert its therapeutic effects by inducing ferroptosis in FGPs cells.

Key words: ferroptosis, network pharmacology, organoids, fundic gland polyps, Weifuchun

LI Yue, DENG Jinyan, PI Shanshan, ZHANG Yingjuan, ZHAO Dan, GUO Yi, YE Yong’an, ZAO Xiaobin, DU Hongbo. Weifuchun (胃复春 ) exerts therapeutic effects on gastric fundic gland polyps by promoting ferroptosis[J]. Journal of Traditional Chinese Medicine, 2025, 45(3): 618-627.

Figures/Tables 6

Figure 1 Morphology and histopathology features of FGP-PDOs A: morphology of primary, passaged, and thawed organoids under bright-field microscopy at different magnifications. A1-A3: bright-field microscope × 50; A4-A6: bright-field microscope × 100. B: HE staining of the three constructed organoid lines was consistent with that of the primary tissue. B1-B3: FGPs; B4-B6: FGPs-PDO. Primary organoids group: A1, A4; passage 4 (P4) organoids group: A2, A5; organoids at passage 1 after thawed group: A3, A6. Patient 1: B1, B4; Patient 2: B2, B5; Patient 3: B3, B6. Scale bar: 100 μm. FGPs: fundic gland polyps; PDO: patient-derived organoids.HE: hematoxylin-eosin staining.

Figure 2 Effect of WFC on FGPs-PDO cell activity A: bright-field images of three organoid lines in the control group at 0 and 72 h. A1-A3 are bright-field images at 0 h after drug administration; A4-A6 are bright-field images at 72 h after drug administration. A1, A4: Organoid 1; A2, A5: Organoid 2; A3, A6: Organoid 3; A7: the statistical analysis of the average organoid area at 0, 72 h. B: bright-field images of three organoid lines treated with 10 μM sorafenib at 0, 72 h. B1-B3 are bright-field images at 0 h after drug administration; B4-B6 are bright-field images at 72 h after drug administration. B1, B4: Organoid 1; B2, B5: Organoid 2; B3, B6: Organoid 3; B7: the statistical analysis of the average organoid area at 0 and 72 h under sorafenib treatment. C: bright-field images of three organoid lines treated with 250 μg/mL WFC at 0 and 72 h. C1-C3 are bright-field images at 0h after drug administration; C4-C6 are bright-field images at 72 h after drug administration. C1, C4: Organoid 1; C2, C5: Organoid 2; C3, C6: Organoid 3; C7: the statistical analysis of the average organoid area at 0 and 72 h under WFC treatment. A: the control group; no treatment; B: the sorafenib group; treated with 10 μM sorafenib for 72 h; C: the WFC group: treated with 250 μg/mL WFC for 72 h. Scale bar: 100 μm, FGPs: fundic gland polyps; PDO: patient-derived organoids. The unpaired t-test was used to evaluate the differences between the two groups. Data are presented as mean ± standard error of mean (n = 3). aP < 0.001, compared with 0 h.

Table 1 Active components of WFC

Serial No.	Name	Source plant	Serial No.	Name	Source plant
XCC1	Ferulic acid	Herba seu Radix Isodi Amethystoidis	XCC11	Dibutyl terephthalate	Herba seu Radix Isodi Amethystoidis
XCC2	Caffeic acid	Herba seu Radix Isodi Amethystoidis	XCC12	Quercetin	Herba seu Radix Isodi Amethystoidis
XCC3	Lariciresinol	Herba seu Radix Isodi Amethystoidis	ZQ1	Hesperetin	Fructus Aurantii Submaturus
XCC4	Glaucocalyxin A	Herba seu Radix Isodi Amethystoidis	A	beta-sitosterol	Fructus Aurantii Submaturus, Panax ginseng
XCC5	Melissoidesin U	Herba seu Radix Isodi Amethystoidis	ZQ2	naringenin	Fructus Aurantii Submaturus
XCC6	Pomiferin F	Herba seu Radix Isodi Amethystoidis	ZQ3	nobiletin	Fructus Aurantii Submaturus
XCC7	Angustifolin	Herba seu Radix Isodi Amethystoidis	HS1	Dioctyl Phthalate	Panax Ginseng Rubra
XCC8	Glaucocalyxin B	Herba seu Radix Isodi Amethystoidis	HS2	(6Z,10E,14E,18E)-2,6,10,15, 19,23 hexa-methyltetra-cosa-2,6,10,14,18, 22-hexaene	Panax Ginseng Rubra
XCC9	Melissoidesin M	Herba seu Radix Isodi Amethystoidis	HS2		Panax Ginseng Rubra
XCC10	Melissoidesin O	Herba seu Radix Isodi Amethystoidis	HS3	ginsenoside rh2	Panax Ginseng Rubra

Figure 3 WFC's Co-targets and Interaction Network with FGPs A: venn diagram showing the intersection of FGPs disease genes and WFC target genes; B: network diagram illustrating the drug-active ingredient-intersection target relationship between FGPs and WFC; C: PPI network of WFC-FGPs-related targets; D: bubble chart displaying the top 10 enriched pathways from KEGG pathway. WFC: Weifuchun; FGPs: undic gland polyps; PPI: protein-protein interaction; KEGG: Kyoto Encyclopedia of Genes and Genomes.

Table 2 Top 10 active components by degree value

Active Component	Degree Value	Betweenness Centrality	Closeness Centrality
Quercetin	53	0.25	0.42
Glaucocalyxin B	46	0.17	0.41
Melissoidesin U	45	0.15	0.41
Melissoidesin O	42	0.16	0.40
Hesperetin	42	0.17	0.37
Glaucocalyxin A	40	0.12	0.40
Angustifolin	28	0.08	0.37
Melissoidesin M	28	0.08	0.37
Dioctyl Phthalate	24	0.12	0.36
beta-sitosterol	18	0.10	0.34

Figure 4 Organoid morphology and mRNA expression of key ferroptosis-related genes after WFC treatment A: after 2 d of organoid passaging, drug interventions were performed, followed by bright-field morphological observations at 0 and 72 h. A1-A5 are bright-field images at 0 h after drug administration; A6-A10 are bright-field images at 72 h after drug administration. B: mRNA expression levels of SLC7A11 in the control, Erastin, and WFC groups after 72 h of drug treatment. C: mRNA expression levels of ACSL4 in the control, Erastin, and WFC groups after 72 h of drug treatment. D: mRNA expression levels of SLC7A11 in three organoid lines after 72 h of WFC treatment. E: mRNA expression levels of ACSL4 in three organoid lines after 72 h of WFC treatment. F: mRNA expression levels of ALOX5B in three organoid lines after 72 h of WFC treatment. the control group: A1, A6, treated with 0.1% DMSO; Erastin group: A2, A7, treated with the 5 μM Erastin dissolved in DMSO; Fer-1 + erastin group:A3, A8, the group pretreated with 10 μM Fer-1 for 24 h followed by Erastin treatment; WFC group: A4, A9, the group treated with 250 μg/mL WFC dissolved in DMSO; Fer-1+WFC group: A5, A10, the group pretreated with 10 μM Fer-1 for 24 h followed by WFC treatment. Scale bar: 100 μm. WFC: Weifuchun; DMSO: dimethyl sulfoxide; Fer-1: Ferrostin-1. The unpaired t-test was used to evaluate the differences between the two groups. Data are presented as mean ± standard error of mean (n = 3). Compared with control, aP < 0.0001, bP < 0.05, cP < 0.001.

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