体外胰岛素抵抗细胞模型的建立及在药物筛选中的应用

The establishment of insulin resistant model in vitro and preliminary application for screening drugs

目的在体外建立肝胰岛素抵抗细胞模型和脂肪细胞胰岛素抵抗模型,并初步应用于中药有效成分的胰岛素抵抗活性筛选中。方法采用高胰岛素诱发HepG2细胞建立肝胰岛素抵抗模型,地塞米松诱发3T3-L1脂肪细胞建立脂肪细胞胰岛素抵抗模型,研究不同药物对胰岛素抵抗模型细胞葡萄糖消耗的影响。结果将HepG2细胞置于10-6mol·L-1的胰岛素中36h,HepG2细胞对胰岛素的抵抗作用最为明显,该特性可维持48h,但未发现细胞形态学变化;地塞米松作用3T3-L1脂肪细胞后,96h空白组与模型组葡萄糖消耗量差值最大,此时为胰岛素抵抗的最高状态;脂肪细胞胰岛素抵抗模型成立后,撤掉地塞米松,胰岛素抵抗细胞模型能够在24h内稳定。某些中药提取成分(如水苏糖、小檗碱和人参皂苷等)能促进HepG2胰岛素抵抗模型和3T3-L1脂肪细胞胰岛素抵抗模型的葡萄糖消耗。结论高胰岛素诱导培养法可以复制出稳定可靠的肝胰岛素抵抗细胞模型;地塞米松诱导3T3-L1脂肪细胞也可建立稳定的脂肪细胞胰岛素抵抗模型。

Aim To establish an in vitro insulin resistant model of HepG2 ceils and 333-L1 adipocyte and to screen drug in vitro. Methods The insulin resistant models of HepG2 and 333-L1 adipocyte were induced by high concentration of insulin and by dexamethasone, respectively. The glucose consumption of ceils was detected after administration with different drugs.Results In the concentration of 10^-6 mol·L^-1 insulin for 36 hours,HepG2 cells were resistant to insulin and the insulin resistance was maintained for 48 hours without change of cell morphology. After 3T3-L1 adipocyte insulin resistance was induced by dexamethasone, the maximal difference of glucose consumption between the blank control and insulin resistant model group was observed at 96h after dexamethasone administration, but the insulin resistant status had only been maintained for 24 hours without dexamethasone. The glucose consumptions of insulin re- sistant model of HepG2 and 3r13-L1 adipocyte were promoted by some drugs such as stachyose, berberine and ginsenoside. Con- clusions The insulin resistant model of HepG2 cell and 3r13-L1 adipocyte was successfully established in vitro by high concentration of insulin and by dexamethasone, respectively. It can be used to screen druas for treatment of insulin resistance.