Danggui Jixueteng Decoction protects HS-5 cells against chemotherapy-induced injury via the AKT/FOXO3a pathway to suppress excessive mitophagy and ferroptosis.

ETHNOPHARMACOLOGICAL RELEVANCE: Chemotherapy-induced myelosuppression is a common and severe complication in clinical oncology treatment. Chemotherapeutic agents trigger this condition by damaging bone marrow mesenchymal stem cells (BMSCs) and disrupting hematopoietic microenvironment homeostasis. Danggui Jixueteng Decoction (DJD), a classical blood-tonifying and blood-activating formula, has been confirmed to alleviate post-chemotherapy myelosuppression, but its specific molecular mechanisms in protecting BMSCs remain unclear.

OBJECTIVE

This study aims to clarify the molecular mechanisms whereby DJD-containing serum alleviates chemotherapy-induced BMSCs injury, identify key signaling pathways and targets for its myeloprotection, and provide evidence supporting DJD for treating chemotherapy-induced myelosuppression.

MATERIALS AND

METHODS

UHPLC-Orbitrap-MS analyzed DJD's chemical constituents, including blood-entry prototypes and metabolites. A carboplatin (CBP) -induced cell injury model was established. Different concentrations of DJD-containing serum were applied. Functional, proteomic, Western blot, molecular docking, cellular thermal shift assay (CETSA) and molecular dynamics simulation (MD) were used for detection and verification.

RESULTS

107 chemical components and 38 blood-entry components of DJD were identified. DJD-containing serum concentration-dependently reversed the aforementioned damaging effects of CBP, significantly improved mitochondrial function, suppressed excessive mitophagy and ferroptosis, and effectively restored AKT/FOXO3a pathway signaling function. Molecular docking, CETSA and MD validated that tanshinol B stably binds to FOXO3a and enhances its thermal stability.

CONCLUSION

This study demonstrates that DJD-containing serum protects BMSCs and maintains hematopoietic microenvironment homeostasis by targeting the AKT/FOXO3a signaling pathway, thereby repairing its abnormally inhibited state, suppressing CBP-induced excessive mitophagy, and inhibiting subsequent ferroptosis. This study provides novel insights and experimental evidence for targeted intervention of chemotherapy-induced bone marrow injury using traditional Chinese medicine.