Ginsenoside Rg3 antagonises endometriosis glycolysis via the tripartite motif containing 28 and pyruvate dehydrogenase kinase 4 signalling pathway

BACKGROUND: Endometriosis is a common gynaecological disorder characterised by the ectopic growth of endometrial-like tissue, which can lead to clinical symptoms such as chronic inflammation, pelvic pain, and infertility. Aberrant cellular energy metabolism driven by glycolysis plays a crucial role in the pathogenesis and progression of endometriosis. Ginsenoside Rg3 has demonstrated broad pharmacological potential in cancer, inflammation, and gynaecological diseases. However, its specific regulatory mechanisms in the treatment of endometriosis mediated by abnormal glucose metabolism remain to be further elucidated. PURPOSE: This study focuses on the role and molecular mechanisms of ginsenoside Rg3 in the progression of endometriosis and its regulation of glycolysis. METHODS: The proliferation and migration capabilities of endometriotic cells were assessed using cell biology behaviour assays. Flow cytometry was used to analyse apoptosis and cell cycle progression. Glycolytic function was evaluated by measuring glucose consumption and lactate production. The co-localization and interaction between TRIM28 (Tripartite Motif Containing 28) and PDK4 (Pyruvate Dehydrogenase Kinase 4) were examined using laser confocal microscopy. RNA stability assays were conducted to determine PDK4 mRNA decay rates. RNA immunoprecipitation assays were performed to verify the interaction between TRIM28 and PDK4 mRNA. Additionally, a mouse model of endometriosis was established, and the regulatory effect of Rg3 on disease progression was validated using immunohistochemistry. RESULTS: Through eukaryotic reference transcriptome sequencing, we identified the key glycolytic regulatory factor PDK4 as a potential target gene of Rg3. Concurrently, PDK4 expression was significantly elevated in ectopic endometrial tissues from endometriosis patients. Further investigation revealed that Rg3 downregulated PDK4 protein expression and antagonised endometriosis-related glycolysis by targeting PDK4. Moreover, we identified TRIM28 as a novel interacting protein of PDK4 in endometriosis through mass spectrometry analysis. We subsequently report that Rg3 treatment effectively inhibits TRIM28 and PDK4 binding. Notably, TRIM28 transcriptionally regulates PDK4 mRNA levels, consequently impacting the cellular biological behaviour and glycolysis of endometriosis. Furthermore, TRIM28 expression in ectopic endometrial tissues positively correlate with those of PDK4. CONCLUSION: This study reveals a novel molecular mechanism through which ginsenoside Rg3 antagonises endometriosis, providing a critical theoretical basis and experimental foundation for the development of new targeted therapeutic strategies against this disease.