Piper betle extract inhibits ferroptosis by scavenging oxyradicals, hydroperoxides and inducing NRF2 regulated antioxidants

Abstract

Ferroptosis, an iron catalysed programmed cell death initiated by membrane lipid peroxidation (LPO), is implicated in various degenerative diseases. We screened medicinal/food plants and discovered leaf extract of Piper betle (PB), a popular mouth freshener, as a ferroptosis inhibitor. Compared to vehicle, PB suppressed LPO and inhibited ferroptosis triggered by rotenone or RSL3. Mechanistic studies revealed that PB extract is a powerful lipophilic radical trapping antioxidant and an inducer of Nuclear factor-erythroid 2-Related Factor 2 (NRF2) regulated antioxidant defenses (glutathione/glutathione peroxidase-4 axis). Utilizing diverse free radical systems (hydroxyl, peroxyl, hydroperoxides) and LPO models (serum, LDL), we found that PB extract effectively inhibits LPO by scavenging lipid oxyradicals. LC-HRMS analysis identified hydroxychavicol and polyphenols as the key bioactive constituents in PB extract. In a Drosophila model of rotenone-induced neurotoxicity, PB extract supplementation prevented locomotor deficits and mortality compared to the control diet. In conclusion, PB extract could be developed as a nutraceutical for mitigating ferroptosis-linked disorders.