Abstract: In this study, puerarin (PUE)-loaded liposomes with both mitochondria and brain targeting properties were prepared and evaluated for their protective effects against deoxynivalenol (DON)-induced ferroptosis in neuronal cells. PUE-loaded liposomes (TPP/RVG29-Lipo@PUE (T/R-L@PUE)) co-modified with triphenylphosphine (TPP) cation and rabies virus glycoprotein 29 (RVG29) were prepared by the thin film hydration method. The encapsulation efficiency, in vitro release, microscopic morphology, stability and the mitochondria targeting capability of the liposomes were characterized by various techniques including high-performance liquid chromatography (HPLC), transmission electron microscopy (TEM), and confocal microscopy. The results showed that T/R-L@PUE had an average particle size of 127.20 nm, with a high encapsulation efficiency (70.56%) and drug loading efficiency (9.20%) for PUE, and exhibited excellent mitochondria targeting capacity. Based on these findings, a rat C6 glioma cell model of DON-induced toxicity was established. Compared with the same dosage of free PUE, T/R-L@PUE significantly alleviated DON-induced ferroptosis in C6 cells, indicating that the mitochondria-targeting liposomes effectively improved the bioavailability of PUE. Furthermore, the results showed that T/R-L@PUE alleviated DON-induced ferroptosis in neuronal cells by enhancing cellular antioxidant capacity, regulating iron metabolism, and restoring the solute carrier family 7 member 11 (Slc7a11)-glutathione (GSH)-glutathione peroxidase 4 (GPX4) axis functions. This provides a theoretical basis and technical reference for mitigating the toxic effects of mycotoxins.

Key words: deoxynivalenol; ferroptosis; puerarin; liposomes; mitochondrial targeting