Cell culture and oxygen-glucose deprivation/ reoxygenation (OGD/R)

PC12 cells were seeded onto 6-well plates in a humidified atmosphere at 37 °C with 5% CO₂. PC12 cells were pre-treated with different concentrations of Sal for 24 h before OGD. Subsequently, the culture medium was replaced with DMEM without glucose, and the cells were subjected to hypoxic conditions with 5% CO₂, 94% N₂ and 1% O₂ at 37 °C for 4 h. Then, the cells were fed normal culture medium and returned to the incubator under normoxic conditions for an additional 24 h. The viability of PC12 cells treated with different doses of Sal (1, 10 and 100 μM) was assessed by the CCK-8 assay.