吡格列酮对胰岛素抵抗HepG2细胞模型的药理学评价

目的应用高胰岛素诱导培养HepG2细胞,建立胰岛素抵抗的细胞模型。并探讨吡格列酮对该模型胰岛素敏感性和糖代谢的影响。方法将HepG2细胞置于5×10-7mol.L-1胰岛素培养液中16 h,采用3H-D-葡萄糖参入试验观察高胰岛素对HepG2细胞葡萄糖摄取率的影响。模型建立后,培养液中加入吡格列酮共同孵育,观察吡格列酮对胰岛素抵抗细胞模型葡萄糖摄取率和葡萄糖消耗量的影响。结果高胰岛素诱导培养的HepG2细胞葡萄糖参入率明显低于未用高胰岛素诱导的HepG2细胞(对照细胞)。将高胰岛素诱导培养的HepG2细胞置于不含胰岛素的培养液中60 h,其细胞葡萄糖摄取率仍明显低于对照细胞。含有吡格列酮(浓度为1×10-6~1×10-4mol.L-1)的胰岛素抵抗HepG2细胞的葡萄糖参入率与葡萄糖消耗量明显高于不含吡格列酮的胰岛素抵抗HepG2细胞(P<0.01)。结论将HepG2置于5×10-7mol.L-1胰岛素环境中16 h,该细胞对胰岛素的生物学效应产生抵抗,其胰岛素抵抗状态可维持60 h。该方法较为简便、易行、重复性好、成功率高,可广泛用于胰岛素抵抗的研究。吡格列酮能增加胰岛素抵抗模型细胞的胰岛素敏感性,并能明显改善糖代谢。

Establishment of Insulin-resistant HepG2 cell Model and pharmacological evaluation of pioglitazone

Aim To establish insulin resistant HepG2 cell model induced by high concentration insulin in vitro culture and evaluate the effects of pioglitazone on insulin sensitivity and glucose metabolism in the cell model.Methods HepG2 cells were incubated with 5×10~(-7)mol·L~(-1) insulin for 16 h and insulin-stimulated()~3H-D glucose incorporation was determined.Then,insulin-stimulated glucose incorporation rate which was used to estimate insulin sensitivity of HepG2 cell,and glucose consumption,the amountsof glucose disappeared from the culture medium of HepG2 cells within 24 hours was determined.The insulin resistance cells were incubated with pioglitazone 1×10~(-5) mol·L~(-1).Results It was demonstrated that the incorporation rate of()~3H-D-glucose in insulin-resistant HepG2 cells was decreased significantly than that in the control cells.Insulin-resistant HepG2 cells were free from stimulation for 48 h by insulin,but the incorporation rate of()~3H-D-glucose was lower than that in control cells.Incubation of insulin resistance cells with pioglitazone significantly increased glucose uptake and glucose consumption by the cells compared with that of the control cells(P<0.01).Conclusion HepG2 cells exposed to 5×10~(-7) mol·L~(-1) insulin for 16 h were able to induce a state of insulin resistance and reproduce a insulin-resistant cell model.The state of insulin resistance can be maintained for 48 h.The method is simple,practica and reliable. Pioglitazone can improve the insulin sensitivity and glucose metabolism in the cell model.