To determine reactive oxygen species (ROS) levels, hippocampal tissues were cut into pieces and neurons were isolated using an ophthalmic nipper. The cells were collected after centrifugation at 500 × g for 10 min, washed with PBS, and incubated with dichlorofluorescein diacetate buffer (MedChemExpress, Princeton, NJ, USA) at 37°C for 30 min. Fluorescence intensity was measured using a microplate reader (Thermo-Max, Molecular Devices, Sunnyvale, CA, USA) with excitation/emission at 493/522 nm. The results were expressed as the ratio of ROS content to that of the normal group.
Hippocampal neurons were collected using the aforementioned procedures. Cells were subjected to different treatments according to the instructions of the total glutathione (GSH) detection kit, total superoxide dismutase (SOD) activity detection kit, and malondialdehyde (MDA) detection kit (all from Beyotime). Using a microplate reader, the absorbance at 410 nm was measured to calculate the total GSH content. The absorbance at 560 nm was measured to calculate SOD activity. The absorbance at 532 nm was measured to calculate MDA content.
Hippocampal neurons were collected using the aforementioned procedures. Cells were subjected to different treatments according to the instructions of the total glutathione (GSH) detection kit, total superoxide dismutase (SOD) activity detection kit, and malondialdehyde (MDA) detection kit (all from Beyotime). Using a microplate reader, the absorbance at 410 nm was measured to calculate the total GSH content. The absorbance at 560 nm was measured to calculate SOD activity. The absorbance at 532 nm was measured to calculate MDA content.