|
Objective:Through the cultivation of3T3-L1pre-adipocytes, induction of differentiation to mature, study the influence of different concentration of uric acid on glucose metabolism. By testing the expression of IRS-1and tyrosine phosphorylation in those mature fat cells, to investigate whether high uric acid may be aggravating factors in the defect of the insulin receptor signal transduction pathway.Methods:The experiments were designed for three parts:the first part was the differentiation of cells.3T3-L1pre-adipocytes were incubate with isobutylmenthylxanthine(0.5mmol/1), dexamethasone(1umol/1), insulin(10ug/ml).cells were differentiated into adipocytes as determined by Oil Red O staining. The second part:containing different concentrations (0,2.5,5,7.5,10,15mg/dl) of uric acid intervention in mature fat cells1h,6h,12h,24h,48h collected culture medium, with glucose oxidase method to detect the amount of glucose in the culture medium, fat cells in each group calculated glucose consumption. Part3: Mature fat cells with uric acid intervention48h, the expression of IRS-1protein and the level of tyrpsine phosphorylation of IRS-1protein were detected by western-blot.Results:3T3-L1cells were differentiated into adipocytes as determined by Oil Red O staining.With different concentrations of uric acid (0,2.5,5,7.5,10,15mg/dl) in the medium effect on the mature fat cells, with the extension of time, compared with the control group, there was no significant differences at2.5mg/dl group; at5-15mg/dl groups their glucose consumption decreased obviously,the difference was statistically significant (P<0.05).Different concentrations of uric acid (0,2.5,5,7.5,10,15mg/dl) in the medium acting on the mature fat cells, compared with the control group, the expression of IRS-1there was no difference in experimental group, and the expression of its trosine phosphorylation was significantly decreased, the difference was statistically significant (P<0.05or<0.01).Conclusion:High uric acid can reduce glucose absorption of fat cells which were induced by insulin, affect glucose metabolism of fat cells, which may be one of the causes of insulin resistance。Tyrosine phosphorylation is lower in experimental groups, it indicates that the way of the insulin receptor signal transduction pathway is inhibited may be one of the cause that high uric acid leads to insulin resistance.The thesis comprises7figures,2tables and48references
|