Mesenchymal Stem Cells Alleviate Crohn's Disease by Regulating CBS-Mediated Ferroptosis.

BACKGROUND: Mesenchymal stem cells (MSCs) possess immunoregulatory properties and have been shown to alleviate symptoms of Crohn's disease (CD). While ferroptosis has been implicated in the onset of CD, the precise connection between MSCs, CD, and ferroptosis remains unclear. This study sought to explore whether MSCs could alleviate CD by inhibiting ferroptosis. MATERIALS AND METHODS: We screened the hub genes involved in both ferroptosis and CD and validated them in clinical colon biopsies by immunohistochemistry (IHC). MSCs were administered intraperitoneally to mice with colitis induced by trinitrobenzene sulfonic acid (TNBS) enema to evaluate the therapeutic effect. Additionally, a rescue experiment was conducted by co-treating colitis mice with a cystathionine-β-synthase (CBS) inhibitor (aminooxyacetic acid, AOAA) and MSCs to further explore the mechanism of the MSCs-CBS-ferroptosis axis. The expression levels of CBS and ferroptosis-related markers in colonic tissues were assessed by real-time quantitative polymerase chain reaction (RT-qPCR), western blot, and IHC. RESULTS: Bioinformatics and machine learning identified CBS as a hub gene in both CD and ferroptosis. IHC revealed that CBS expression was downregulated in colon tissues from CD patients. MSCs treatment significantly reduced pathological symptoms and intestinal inflammation in colitis mice. MSCs increased CBS expression, which had been diminished in colitis mice. Additionally, MSCs increased glutathione (GSH) and glutathione peroxidase 4 (GPX4) expression, thereby inhibiting ferroptosis activation. In a rescue experiment, co-treatment with AOAA and MSCs exacerbated intestinal inflammation and reversed the elevation of CBS and ferroptosis-related markers. DISCUSSION: This study demonstrates that MSCs alleviate the symptoms of CD by regulating CBS-mediated ferroptosis.