HepG2细胞胰岛素抵抗模型建立及在筛选天然有效多糖中的应用

Establishment of Insulin Resistance Model of HepG2 Cells and Application in Selection of Active Natural Polysaccharides

探讨高糖、地塞米松、胰岛素诱导肝细胞HepG2胰岛素抵抗模型(IR-HepG2)的建立方法,检测杏鲍菇多糖、红枣多糖和枸杞多糖体外改善肝细胞胰岛素抵抗的作用,评估3种多糖的降糖效果.选择不同浓度的地塞米松、胰岛素及两者联合作用,在高糖培养条件下诱导HepG2胰岛素抵抗,MTT法检测不同诱导剂对HepG2细胞活性的影响,葡萄糖氧化酶法测定细胞葡萄糖消耗量,同时检测去除诱导剂后,IR-HepG2细胞模型的稳定性.以3种天然多糖处理IR-HepG2细胞,检测其对细胞活力及胰岛素抵抗的影响.结果表明:在高糖培养条件下,诱导剂为地塞米松(3.75μmol/L)联合胰岛素(0.001μmol/L)可成功诱导IR-HepG2细胞模型,且对细胞活性无显著影响,模型能够在72 h内保持稳定.枸杞多糖具有更好的降糖效果.

To explore the establishment of insulin resistance model of HepG2(IR-HepG2)induced by high glucose,dexamethasone and insulin,to detect the effect of Pleurotus eryngii polysaccharides,Ziziphus jujuba polysaccharides and Lycium barbarum polysaccharides on improving insulin resistance of liver cells in vitro,and to analyze the effect of three polysaccharides on hypoglycemic.Different concentrations of dexamethasone,insulin and their combination were selected to induce the insulin resistance of HepG2 cells in high glucose culture.MTT method was used to detect the effect of different inducers on the activity of HepG2 cells.Glucose oxidase method was used to detect the glucose content in the supernatant of cell culture.At the same time,the stability of IR-HepG2 cell model was detected after removing inducers.IR-HepG2 cells were treated with three kinds of natural polysaccharides at different concentrations.The effects of polysaccharides on cell viability and insulin resistance were detected by MTT and glucose oxidase methods respectively.Dexamethasone(3.75μmol/L)combined with insulin(0.001μmol/L)can successfully induce IR-HepG2 cell model in high glucose culture condition,and has no significant effect on cell activity,and the model can keep stable within 72 hours.Lycium barbarum polysaccharides had better hypoglycemic effects in vitro.