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Ferroptosis is a form of necrosis caused by iron-induced accumulation of lipid hydroperoxide, involving several molecular events, and has been implicated in Parkinson's disease. Gastrodin is a component of Gastrodia elata Blume with strong antioxidant activity. We examined whether gastrodin can prevent H 2 O 2 -induced cytotoxicity in rat glioma cell line C6. For this purpose, C6 cells were pretreated with gastrodin (1, 5, 25 µM) and then exposed to 100 µM H 2 O 2 . Results showed that pretreatment of C6 cells with gastrodin decreased H 2 O 2 -induced lactate dehydrogenase (LDH) release and cell death. Moreover, gastrodin decreased intracellular malondialdehyde (MDA) level, whereas increased glutathione peroxidase (GPX) activity and glutathione (GSH) level after H 2 O 2 treatment. In addition, treatment of deferoxamine (DFO), ferrostatin-1, and liproxstatin-1 abolished ferroptosis induced by H 2 O 2 or erastin pretreatment. Treatment with gastrodin attenuated H 2 O 2 -induced ferroptosis and decreased lipid reactive oxygen species (ROS) (C11-BODIPY) production in C6 cells. Moreover, gastrodin increased the protein expression of nuclear factor erythroid 2-related factor 2 (Nrf2), GPX4, ferroportin-1 (FPN1), and heme oxygenase-1 (HO-1) in C6 cells treated with H 2 O 2 . RSL3, a GPX4 inhibitor, inhibited GPX4 protein level in cells co-treated with gastrodin and 100 µM H 2 O 2 . These findings indicate that gastrodin can inhibit H 2 O 2 -induced ferroptosis through its antioxidative effect in C6 cells.

Gastrodin Inhibits H<sub>2</sub>O<sub>2</sub>-Induced Ferroptosis through Its Antioxidative Effect in Rat Glioma Cell Line C6