4.5. High Glucose and Fatty Acids-Induced Insulin Resistance In Vitro

Human liver carcinoma cell HepG2 was cultured in low glucose (5.5 mM) DMEM medium supplemented with 10% FBS and 1% antibiotics in a 96-well plate for 24 h. Cell culture medium was changed to FG conditioned medium containing 25 mM glucose and 0.5 mM palmitic acid and oleic acid (molar ratio 1:2) with or without polysaccharides for 24 h. Subsequently, the cells were treated with 1 nM insulin for 12 h in serum-free low-glucose DMEM medium. Cells were collected for glucose uptake measured by an assay kit (Applygen, Beijing, China) using a microplate reader (-Infinite F50, Tecan, Switzerland).