GOD-POD法在3T3-L1脂肪细胞胰岛素抵抗评价中的应用

GOD-POD assay for evaluating insulin resistance of 3T3-L1 adipocytes

目的:通过葡萄糖氧化酶-过氧化物酶(Glucose oxidase-peroxidase,GOD-POD)法检测脂肪细胞葡萄糖消耗,建立简便、经济的脂肪细胞胰岛素敏感性的评价方法。方法:诱导分化成熟的3T3-L1脂肪细胞,分别采用高浓度葡萄糖(25 mmol/L)、地塞米松(Dexamethasone,DEX)(1.0μmol/L)诱导脂肪细胞产生胰岛素抵抗(Insulin resistance,IR),GOD-POD法测定糖浓度,动态观察诱导过程中细胞糖消耗,并计算胰岛素敏感性指数(Insulin sensitivity index,ISI);同时应用RT-PCR检测脂联素(Adiponectin)mRNA表达。结果:通过该方法检测,可以反映脂肪细胞对葡萄糖的摄取利用,以胰岛素刺激下较基础状态细胞葡萄糖消耗的增加量,计算出ISI,能够反映细胞对胰岛素的敏感性及抵抗程度。结论:GOD-POD法能够准确测定细胞总体葡萄糖消耗,可用于脂肪细胞糖摄取利用与IR评价,此方法操作简便、经济易行。

Objective：To establish a simple and economical method for evaluating the insulin resistance of adipocyte with Glucose oxidase-peroxidase（GOD-POD） assay in vitro.Methods：3T3-L1 cells were induced into mature adipocytes which were stimulated with glucose（25 mmol/L） and DEX（1.0 μmol/L） respectively to induce insulin resistance.Glucose concentration was measured with GOD-POD assay during the induction,the glucose consumption dynamic viewed and the insulin sensitivity index（ISI） calculated.And the mRNA expression of Adiponectin was tested by RT-PCR.Results：The glucose metabolism of adipocytes was detected with this method.The insulin sensitivity and resistance levels of adipocytes were reflected by ISI calculated by the increase of insulin-stimulated glucose consumption.Conclusion：Glucose consumption can be accurately detected with GOD-POD assay which proves to be a simple,easy and economical method to evaluate insulin resistance of adipocytes.