Ferroptosis-Regulated Osteoblasts Under Compression Via GPX4/RUNX1 Pathway

Abstract

Background: The adaptive remodeling of alveolar bone ensures the effectiveness and stability of orthodontic tooth movement (OTM), but how to accurately regulate the remodeling of alveolar bone is the focus of orthodontic research. Ferroptosis plays an important role in bone remodeling. Methods: To determine the generation and mechanism of ferroptosis during orthodontic treatment, we used immunohistochemical method to detect the expression of ferroptosis related proteins GPX4 and ACSL4. We performed proteomic analysis of GPX4 knockdown of MC3T3-E1 and predicted the potential transcription factor. Dual luciferase assay and Chip-qPCR assay were used to test the GPX4 promoter region. RUNX1 level was regulated through in vitro and in vivo experiments. Results: Compressive force significantly reduced GPX4 in alveolar bone, indicating that compressive force induced ferroptosis. JASPAR database displayed that RUNX1 may be a key transcription factor for glutathione peroxidase 4 (GPX4) during OTM. Chip -qPCR indicated that RUNX1 binds to the GPX4 promoter region, and RUNX1 agonist kartogenin could effectively save the osteogenic loss caused by the reduction of GPX4. By regulating GPX4 levels, GPX4 increasing could not reverse the decrease of RUNX1. Finally, by examining changes in osteogenic levels in MC3T3-E1 cells under compressive force after kartogenin treatment, we found that the increase of RUNX1 contributed to the reduction of osteogenic levels due to compressive force. Conclusion: Our findings suggest that the RUNX1/GPX4 signaling axis may be a potential therapeutic target during orthodontic tooth movement that can promote alveolar bone remodeling by inhibiting ferroptosis in OTM.