ObjectiveThis study aims to investigate the molecular mechanism by which Guizhi Fulingwan （GFW） inhibits ferroptosis in endometriosis （EMT） through the regulation of the hypoxia inducible factor-1α/heme oxygenase 1 （HIF-1α/HO-1） signaling pathway. MethodsMachine learning was employed to identify ferroptosis-related biomarkers associated with EMT. Network pharmacology was utilized to identify the active components of GFW and its potential therapeutic targets against EMT， including core targets. Functional enrichment analysis was conducted to explore the biological processes， molecular functions， cellular components， and signaling pathways associated with the potential targets. An EMT rat model was established via autologous transplantation. Thirty female Sprague-Dawley （SD） rats were randomly divided into five groups： sham-operated， model， positive control （dienogest at 0.2 mg·kg^-1）， low-dose GFW （2.5 g·kg^-1）， and high-dose GFW （5 g·kg^-1）. After modeling， the rats received their respective treatment by oral gavage for 28 consecutive days， while the sham and model groups received equal volumes of distilled water. Serum and ectopic endometrial tissues were collected. Hematoxylin and eosin （HE） staining was employed to evaluate morphological alterations in ectopic lesions. Quantitative real-time polymerase chain reaction （Real-time PCR） and Western blot were conducted to assess mRNA and protein expression of HIF-1α， HO-1， glutathione peroxidase 4 （GPX4）， spermidine/spermine N1-acetyltransferase （SAT1）， and prostaglandin-endoperoxide synthase 2 （PTGS2）. Tissue levels of malondialdehyde （MDA）， glutathione （GSH）， and ferrous iron （Fe²⁺） were quantified using commercial assay kits. Serum levels of interleukin-6 （IL-6） and transforming growth factor-β₁ （TGF-β₁） were measured via enzyme-linked immunosorbent assay （ELISA）. ResultsFive ferroptosis-related biomarkers in EMT were identified： ALOX12， CHAC1， SAT1， AST1， and HO-1. Network pharmacology analysis revealed 42 active components of GFW and 192 potential therapeutic target genes related to EMT treatment， with FOS， JUN， HO-1 identified as core targets. Functional enrichment analysis indicated that the potential targets were primarily involved in oxidative stress response and reactive oxygen species metabolism and were enriched in the HIF-1 signaling pathway. Compared to the sham-operated group， the model group exhibited significant increases in both mRNA and protein expression of HIF-1α， HO-1， and PTGS2， as well as elevated tissue levels of Fe²⁺ and MDA. Conversely， GSH levels and the expression of GPX4 and SAT1 were markedly reduced， and serum levels of IL-6 and TGF-β₁ levels were significantly higher （P<0.01）. Compared with the model group， all GFW-treated groups showed significant downregulation of HIF-1α and HO-1， reduced Fe²⁺ levels， and downregulated expression of MDA， PTGS2， IL-6， and TGF-β₁. Meanwhile， GSH， GPX4， and SAT1 expression levels were significantly increased （P<0.05， P<0.01）， effectively ameliorating iron overload and oxidative stress， thereby demonstrating therapeutic efficacy in EMT， with the high-dose GFW demonstrating the most pronounced therapeutic effects. ConclusionGFW exerts therapeutic effects on endometriosis by regulating the HIF-1α/HO-1 signaling pathway to rectify iron metabolism disorders and attenuate free iron-induced oxidative damage. It upregulates the antioxidative defense system to inhibit lipid peroxidation cascades and modulates inflammatory cytokine networks. These effects collectively disrupt the pathological interaction between ferroptosis and chronic inflammation， providing a novel theoretical foundation for the clinical application of GFW in EMT treatment.

Objective To investigate the mechanism of Wenjing Decoction in treating endometriosis(EMT)using bioinformatics methods and in vitro experiments.Methods The active components and corresponding targets of Wenjing Decoction were obtained from the TCMSP database,while EMT-related targets were identified using the GEO database.Functional enrichment analysis was conducted on the targets to predict core targets for treating EMT with Wenjing Decoction.Molecular docking was performed on core targets-drug ligands,and in vitro experiments validated the findings.Results Through screening the TCMSP database,117 active components of Wenjing Decoction were identified,corresponding to 248 targets;5 312 EMT-related differential genes were gathered from GEO database,identifying 97 potential targets of Wenjing Decoction for treating EMT,with core targets being IL6,TNF and EGFR.Functional enrichment analysis of EMT differential genes showed enrichment in pathways such as neuroactive ligand-receptor interaction,MAPK signaling pathway,endocytosis,calcium signaling pathway,autophagy and PI3K-Akt signaling pathway.Molecular docking showed that IL6,TNF,EGFR bind stably to their corresponding drug ligands.In vitro experiments indicated that Wenjing Decoction could inhibit the PI3K/Akt/mTOR pathway,promote LC3 Ⅰ to LC3 Ⅱ conversion,enhance the expression of Beclin-1,and reduce P62 expression.Moreover,Wenjing Decoction could hinder the expression of the endometriosis-specific biomarker CA125,decrease EGFR,IL-6 and TNF-α expressions in ectopic endothelial cells,inhibiting proliferation.Conclusion Wenjing Decoction can treat EMT through multiple pathways and targets,with the key mechanism being the reversal of autophagy inhibition via down-regulating of the PI3K/Akt/mTOR pathway.

Objective To investigate the mechanism of Wenjing Decoction in treating endometriosis(EMT)using bioinformatics methods and in vitro experiments.Methods The active components and corresponding targets of Wenjing Decoction were obtained from the TCMSP database,while EMT-related targets were identified using the GEO database.Functional enrichment analysis was conducted on the targets to predict core targets for treating EMT with Wenjing Decoction.Molecular docking was performed on core targets-drug ligands,and in vitro experiments validated the findings.Results Through screening the TCMSP database,117 active components of Wenjing Decoction were identified,corresponding to 248 targets;5 312 EMT-related differential genes were gathered from GEO database,identifying 97 potential targets of Wenjing Decoction for treating EMT,with core targets being IL6,TNF and EGFR.Functional enrichment analysis of EMT differential genes showed enrichment in pathways such as neuroactive ligand-receptor interaction,MAPK signaling pathway,endocytosis,calcium signaling pathway,autophagy and PI3K-Akt signaling pathway.Molecular docking showed that IL6,TNF,EGFR bind stably to their corresponding drug ligands.In vitro experiments indicated that Wenjing Decoction could inhibit the PI3K/Akt/mTOR pathway,promote LC3 Ⅰ to LC3 Ⅱ conversion,enhance the expression of Beclin-1,and reduce P62 expression.Moreover,Wenjing Decoction could hinder the expression of the endometriosis-specific biomarker CA125,decrease EGFR,IL-6 and TNF-α expressions in ectopic endothelial cells,inhibiting proliferation.Conclusion Wenjing Decoction can treat EMT through multiple pathways and targets,with the key mechanism being the reversal of autophagy inhibition via down-regulating of the PI3K/Akt/mTOR pathway.