Penicillin streptomycin p s

The human monocytic leukemia cell line (THP-1), the human-type II cell alveolar epithelial cells (A549) and human umbilical vein endothelial cells (HUVEC) were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). THP-1 cells were cultured in RPMI 1640 culture medium (Servicebio, Wuhan, China). A549 cells were cultured in F12K medium and HUVEC was cultured in DMEM medium with high glucose. All the culture media were supplemented with 10% fetal bovine serum (FBS; Gibco, Waltham, MA, USA) and 1% penicillin/streptomycin (P/S) (Biosharp, Hefei, China). Cells were maintained at 37 °C, 5% CO2 humidified incubator. For Nrf2 knockdown assay, the THP-1 cells were transfected with Nrf2 siRNA (5′-AUUGAUGUUUCUGAUCUAUCACUTT-3′) using Lipofectamine 2000 (Invitrogen, Waltham, MA, USA) according to the instruction.

The human monocytic leukemia cell line THP-1 were obtained from the American Type Culture Collection (ATCC, Manassas VA, USA) and was cultured in RPMI 1640 culture medium (Servicebio, China) supplemented with 10% fetal bovine serum (FBS; Gibco, US) and 1% penicillin/streptomycin (P/S) (Biosharp, China), and maintained at 37 °C, 5% CO₂ humidified incubator. For the siRNA transfections, the cells were transfected with Nrf2 siRNA (5′-AUUGAUGUUUCUGAUCUAUCACUTT-3′). Lipofectamine 3000 (Invitrogen, USA) was used for plasmid transfections according to the manufacturer’s instructions.

Oleanolic acid was extracted from Ligustrum lucidum by ultrasonic-assisted technology. Dulbecco’s modified Eagle’s medium (DMEM) was obtained from biological industries (Shanghai, China). A CCK-8 kit, nonessential amino acids (NEAAs), trypsin−EDTA, PBS and penicillin−streptomycin (PS) were obtained from Biosharp (Hefei, China). Cell cycle and apoptosis analysis kits were obtained from Beyotime (Shanghai, China). An Annexin V-FITC/PI apoptosis detection kit was obtained from Vazyme (Nanjing, China).