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目的体外用地塞米松诱导培养法建立及鉴定肝胰岛素抵抗细胞模型。方法将HepG2细胞置于含不同浓度地塞米松培养基中培养24 h,用含胰岛素的新鲜培养基刺激24 h,采用葡萄糖氧化酶-过氧化物酶(glucose oxidase-peroxide enzyme,GOD-POD)法检测细胞对葡萄糖的消耗情况,蒽酮法检测细胞内糖原含量,利用Western blot检测细胞内胰岛素受体底物-2表达的变化。结果用地塞米松处理HepG2细胞24 h后,细胞对胰岛素的敏感性显著降低,1.0、3.3、10.0μmol/L地塞米松组葡萄糖消耗分别为(0.637±0.018)、(0.535±0.018)、(0.471±0.011)mmol/L(P<0.05,P<0.01),细胞内糖原含量分别为(2.83±0.13)、(2.42±0.10)、(2.23±0.06)μmol/mg(P<0.05,P<0.01),胰岛素受体底物-2表达量均下降(P<0.05,P<0.01)。结论体外地塞米松诱导培养法可以成功建立肝胰岛素抵抗细胞模型,此细胞模型出现胰岛素抵抗的机制可能与IRS-2表达的下调有关。
Objective To establish and identify hepatic insulin resistance cell model with dexamethasone in vitro. Methods HepG2 cells were cultured in dexamethasone medium containing different concentrations for 24 h and stimulated with fresh medium containing insulin for 24 h. Glucose oxidase-peroxidase (GOD-POD) The glucose consumption was detected by the method of cell counting, the intracellular glycogen content was detected by the anthrone method, and the expression of insulin receptor substrate-2 was detected by Western blot. Results Dexamethasone treatment of HepG2 cells for 24 h, the cell sensitivity to insulin was significantly reduced, 1.0,3.3,10.0μmol / L dexamethasone group glucose consumption (0.637 ± 0.018), (0.535 ± 0.018), (0.471 ± 0.011) mmol / L (P <0.05, P <0.01). The contents of intracellular glycogen were (2.83 ± 0.13), (2.42 ± 0.10) and (2.23 ± 0.06) μmol / 0.01), while the expression of insulin receptor substrate-2 decreased (P <0.05, P <0.01). Conclusion In vitro dexamethasone-induced culture successfully established hepatic insulin resistance cell model. The mechanism of insulin resistance in this cell model may be related to the down-regulation of IRS-2 expression.