20-Deoxyingenol alleviates ferroptosis by activating transcription factor EB in spinal cord injury

Background Spinal cord injury (SCI) lacks effective treatments. Ferroptosis contributes to SCI pathology. We investigated the therapeutic potential of 20-deoxyingenol (20-DOI), a natural diterpene, focusing on its role in inhibiting ferroptosis. Methods We used ventral spinal cord 4.1 motor neurons in vitro , challenged with H 2 O 2 or erastin. Assessments included cell viability, proliferation, autophagy, lysosomal function, and ferroptosis. Transcription factor EB (TFEB) knockdown validated its involvement. In vivo , a mouse SCI model assessed functional recovery (Basso mouse scale score), tissue damage, and ferroptosis markers. Results 20-DOI restored cell viability and proliferation, enhanced autophagy and lysosomal activity. It suppressed ferroptosis, reducing lipid peroxidation and reactive oxygen species, and preserving mitochondrial function. These benefits required TFEB; its knockdown abolished the protection and reduced NRF2/GPX4 levels. In mice, 20-DOI improved motor function, preserved neurons, and attenuated ferroptosis. Conclusion 20-DOI promotes recovery after SCI by activating TFEB to inhibit ferroptosis. Our work identifies TFEB as a key target and 20-DOI as a promising therapeutic agent.