2.5. Cell Culture
hyperlipidemia
HYPERLIPIDEMIA
HepG2 cells were obtained from Procell Life Science & Technology (Wuhan, China) and cultured in Dulbecco’s modified eagle medium with 10% fetal bovine serum, 100 U/mL penicillin, and 100 µg/mL streptomycin at 37 °C in 5% CO2. To establish an in vitro hyperlipidemia model, HepG2 cells were treated with free fatty acids (FFA), a mixture of oleate and palmitate with a final concentration of 750 µmol/L, and incubated with or without resveratrol (10 μmol/L) for 48 h. Uric acid levels in culture supernatants were measured by Mindray BS-830 automatic biochemical analyzer (Shenzhen, China). Intracellular XO activity was analyzed and normalized by the total protein measured by commercial kits (Cat NO. PC0020, Solarbio, Beijing, China). The protein expression of XO was analyzed by Western blot with primary antibodies against GAPDH (Cat NO. K200057M, Solarbio, Beijing, China) and XO (Cat NO. ab133268, Abcam, Cambridge, UK).