游离脂肪酸对3T3 L1脂肪细胞糖代谢影响的研究.pdfVIP

Abstract Objective: Through the cultivation of 3T3-L1 preadipocytes, and induction of differentiation to mature study of free fatty acids (FFA) on glucose metabolism. By etecting the 3T3-L1 preadipocytes during differentiation and maturation of intracellular reactive oxygen species (ROS) concentration, intracellular free radical reactions. Methods: Cultured 3T3-L1 preadipocytes by IBMX, insulin, induced by the combination of dexamethasone, cells from the spindle into a big, round and full of lipid droplets in fat cells, that cell differentiation. Oil red O staining and compared the morphological changes in the structure. Through different FFA (LPS, EPA, SA, PA) intervention mature fat cells, different cells with MTT assay of free fatty acids stimulate cell for 48 hours, the survival rate of fat cells. At the same time, containing FFA (LPS, EPA, SA, PA) in DMEM intervention in mature adipocytes 6h, 12h, 24h, 48h collected culture medium, glucose oxidase fat cells in each group calculated glucose consumption. Fat cells was detected by different time of reactive oxygen species concentration. Mature fat cells, detected by Western blot in each group at different times after the intervention FFA cell AMPK, GLUT4 protein content. Results: ① induced differentiation of 3T3-L1 pre-pre-fat cells, spindle-shaped cytoplasm without lipid droplets, showed fibroblast-like morphology of cells to contact inhibition, the stagnation in the growth of; induced to differentiate 2 days rounded morphology, but no lipid droplets in cytoplasm appear; induced differentiation of the first 4 days, cells become greatly changed round, a small amount of lipid droplets in the cytoplasm around the nucleus appears in the oil red O staining of fat cells can be observed in cell pulp