Abstract: AIM: To investigate the protective mechanism of gastrodin against hydrogen peroxide (H2O2) damage in PC12 cells. METHODS: PC12 cells were treated with H2O2 to establish injury model, MTT assay was used to detect cell viability to establish the best suitable concentration of the model and the optimal treatment concentration of gastrodin were determined. Biochemical methods were used to analyze MDA, GSH and SOD activity. Flow cytometry were used to detect the change of ROS. Inverted transmission electron microscopy was used to observe PC12 cell morphology. Western blot was used to analyze the expression of p53, GPX4 protein. RESULTS: MTT assay showed that 50 μmol/L H2O2 inhibited PC12 cells after 48 h of treatment, while 1, 5, 25 μmol/L gastrodin cells reduced the inhibition of H2O2 on PC12 cells. MDA, GSH and SOD results showed that gastrodin reduced MDA level and increased SOD and GSH level after H2O2 treatment. Flow cytometry detection showed that gastrodin pretreatment reduced the content of intracellular reactive oxygen species after H2O2 injury. Transmission electron microscopy showed that the mitochondrial ridge was thicker and the mitochondrial volume was smaller in the 50 μmol/L H2O2 group than in the normal group, while the mitochondrial morphology tend to normal after gastrodin preconditioned PC12 cells. Western blot results showed that gastrodin could increase GPX4 protein expression and decrease p53 protein expression after H2O2 injury. CONCLUSION: H2O2 can cause ferroptosis by increasing the level of oxidative stress in PC12 cells, and gastrodin pre-protection can reduce the generation of oxidative stress and prevent ferroptosis which may be related to the up-regulation of GPX4 protein expression and down-regulation of p53 protein expression.

Key words: gastrodin, ferroptosis, H2O2, oxidative stress, neurodegenerative disease