论文部分内容阅读
目的 探讨高浓度胰岛素和高浓度葡萄糖共同作用对原代培养老鼠脂肪细胞的糖转运活动、细胞内胰岛素信号肽及葡萄糖转运子 4 (GLUT4 )易位的影响。方法 分离的老鼠脂肪细胞在胰岛素 ( 10 4 μU/ml)和不同浓度葡萄糖 ( 5、10、15、2 5mM)培养基中孵育 2 4h ,然后测定糖的转运活动 ;用Westernblot方法测定这些细胞内胰岛素受体底物 (IRS) 1/ 2、肌醇磷脂 - 3 -激酶 85亚单位 (p85)、蛋白激酶B(PKB)和GLUT4的蛋白表达。结果 胰岛素和不同高浓度葡萄糖培养 2 4h ,以一种剂量依赖的方式诱导糖摄取率的减少 ,抑制了IRS1蛋白表达 ,而不影响p85、PKB和GLUT4的表达 ,但抑制了GLUT4易位至质膜 ;胰岛素加正常或不同高浓度葡萄糖治疗 ,均抑制了IRS2蛋白表达。结论 慢性胰岛素治疗诱导的胰岛素抵抗与周围环境中葡萄糖浓度增加有关 ,可能影响了胰岛素受体底物蛋白表达以及GLUT4易位等。
Objective To investigate the effects of high concentration of insulin and high concentration of glucose on glucose transporter activity, intracellular insulin signal peptide and glucose transporter 4 (GLUT4) translocation in primary cultured rat adipocytes. Methods Mouse adipocytes were incubated with insulin (10 4 μU / ml) and different concentrations of glucose (5, 10, 15, 25 mM) for 24 h, then the transport activity of glucose was measured. The intracellular Insulin receptor substrate (IRS) 1/2, phosphoinositide 3-kinase 85 subunit (p85), protein kinase B (PKB) and GLUT4 protein expression. Results Insulin and different concentrations of glucose cultured for 24 hours induced a decrease of glucose uptake rate in a dose-dependent manner, inhibited the expression of IRS1 protein, but did not affect the expression of p85, PKB and GLUT4, but inhibited the GLUT4 translocation to the quality Membrane; Insulin plus normal or different high concentration of glucose treatment, inhibited IRS2 protein expression. CONCLUSION: Insulin resistance induced by chronic insulin therapy is related to the increase of glucose concentration in the surrounding environment, which may affect insulin receptor substrate protein expression and GLUT4 translocation.