Rats wer2.4 mg/kg PE by intraperitoneal administration

SH-SY5Y cells were cultured and maintained in Dulbecco's Modified Eagle Medium (DMEM) solutions (Gibco Laboratories, Grand Island, NY), supplemented with 10% fetal bovine serum (FBS, Gibco Laboratories, Grand Island, NY). Cellswere kept at 37 °C in a humidified 5% CO2/95% O2 incubator. The dissociated cells were seeded in poly-L-lysine-coated plates at a density of 5 × 105/cm2 and cultured in DMEM, supplemented with 10% fetal bovine serum, 100 U/ml penicillin and 0.1 mg/ml streptomycin. The medium was changed twice weekly. After 8 days of culture, the SH-SY5Y cells were randomly divided into seven groups: normal group (no oxygen–glucose deprivation), oxygen–glucose deprivation group (control group) and oxygen–glucose deprivation +PE groups (2.5, 5, 10, 20 and 40 μM). For oxygen–glucose deprivation, the SH-SY5Y cellswere washed twice in glucose-free balanced salt solution (BSS), and incubated in BSS (no glucose) in a humidified chamber filled with 95% N2/5% CO2 for 3 h at 36.5 °C, while the normal group and PE 20 μM group (no oxygen–glucose deprivation) were filled with 5% CO2/95% O2 and incubated in glucose-free BSS. After oxygen–glucose deprivation, the cultureswere replaced into neurobasalmedium and incubated with PE (2.5, 5, 10, 20 and 40 μM) in a CO2 incubator for 12 h.