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目的:采用Urotensin Ⅱ(UⅡ)体外诱导法建立HepG 2细胞胰岛素抵抗模型,并探讨其机制。方法:采用含不同UⅡ浓度的DMEM培养基培养HepG 2细胞,最后半小时与1×10-7mol/L insulin共孵育,葡萄糖-己糖激酶法检测不同时点培养基中葡萄糖含量,以HepG 2细胞蛋白进行定量,确定胰岛素抵抗最佳作用浓度及时间;多功能酶标仪检测UⅡ诱导hepG 2细胞内活性
OBJECTIVE: To establish a model of insulin resistance in HepG2 cells induced by Urotensin Ⅱ (UⅡ) in vitro and its mechanism. Methods: HepG 2 cells were cultured in DMEM medium containing different concentrations of UⅡ, incubated with 1 × 10-7mol / L insulin for half an hour at a final concentration of 1 × 10-7mol / L insulin. Glucose-hexokinase method was used to detect the glucose content in medium at different time points. HepG2 Cell protein was quantified to determine the best concentration of insulin resistance and time; multi-function microplate reader to detect heparin II induced intracellular activity