Studies On Effects Of Free Fatty Acid On Glucose Metabolism In 3t3-l1 Adipocytes

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:â‘ 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 in the lipid droplets stained red; the first 6 days, further changes big changes round fat cells, small lipid droplets increased significantly, gathered around the nucleus, start with "Ring Ring"-like structure; induced differentiation of the first 8 days,90% cell differentiation and maturation are more lipid droplet accumulation, the small lipid droplets merging to form large lipid droplets.â‘¡with LPS, EPA, SA, PA in the medium effect on mature adipocytes, with the time, significantly inhibit the fat cells to absorb glucose (P<0.01).â‘¢3T3-L1 preadipocytes in the induction of the first 8 days, the largest number of cells, the highest activity, induced from day 0 to day 8 in the process of reactive oxygen species concentration in fat cells gradually increased to peak on day 8. To induction, as cells reduce their activity, cells were aging at the same time, fat cells gradually decreased the concentration of reactive oxygen species.â‘£with LPS, EPA, SA, PA in the medium acting on the mature fat cells, with the time, fat cells AMPK, GLUT4 protein content decreased.Conclusions:Fat cells of reactive oxygen species concentration can reflect the state of cell survival, free fatty acid can induce the molecular mechanism of insulin resistance may be through the insulin signaling pathway activated protein kinase (AMPK), thereby affecting the expression of GLUT4 protein to fat cell glucose uptake rate may affect glucose metabolism in fat cells.