HepG2细胞胰岛素抵抗模型的建立及其应用

发布时间：2018-07-11 14:38

本文选题：HepG细胞 + 胰岛素抵抗　；参考：《核农学报》2017年09期

【摘要】：为建立方便、可靠、重复性好的胰岛素抵抗细胞模型,研究胰岛素受体底物-2(IRS-2)蛋白磷酸化与胰岛素抵抗的关系以及不同铬制剂对细胞葡萄糖代谢的作用。试验采用高胰岛素、高糖联合诱导Hep G2细胞建立胰岛素抵抗模型,通过葡萄糖氧化酶法及噻唑蓝比色法(MTT)分析受试物对细胞葡萄糖代谢及活性的影响,优化最佳造模条件;同时,利用蛋白免疫印迹法(WB)分析IRS-2的蛋白表达及其蛋白磷酸化,验证模型的胰岛素抵抗性。在此基础上,将模型应用于分析葡萄糖耐量因子(GTF)及其它铬制剂的降糖效果。结果表明,高胰岛素(10-6mol·L~(-1))和高糖培养基(25 mmol·L~(-1))处理细胞48h,细胞存活率达96%,与对照组相比,葡萄糖消耗量降低了5.7%,IRS-2含量减少31.02%,胰岛素抵抗效果显著,且该胰岛素抵抗模型可稳定维持48 h。相比于吡啶酸铬和三氯化铬,GTF对细胞葡萄糖代谢有显著的改善作用,其最佳作用浓度为1.0μg·m L-1。利用HepG2细胞建立体外胰岛素抵抗模型,方法简单可靠、重复性好,可广泛应用于天然活性物质的降糖功能研究。
[Abstract]:In order to establish a convenient, reliable and reproducible insulin resistance cell model, the relationship between insulin receptor substrate 2 (IRS-2) protein phosphorylation and insulin resistance and the effect of different chromium preparations on glucose metabolism were studied. The insulin resistance model of Hep G2 cells induced by high insulin and high glucose was established. The effects of glucose oxidase and thiazolyl blue colorimetric assay (MTT) on glucose metabolism and activity of Hep G2 cells were analyzed to optimize the optimal model conditions. At the same time, the protein expression and protein phosphorylation of IRS-2 were analyzed by Western blotting (WB) to verify the insulin resistance of the model. On this basis, the model was applied to analyze the hypoglycemic effect of glucose tolerance factor (GTF) and other chromium preparations. The results showed that the survival rate of cells treated with high insulin (10-6mol L-1) and high glucose medium (25 mmol L ~ (-1) for 48 h was 96. Compared with the control group, glucose consumption decreased the content of IRS-2 decreased by 31.02, and the effect of insulin resistance was significant. The insulin resistance model was stable for 48 h. Compared with Chromium picolinate and Chromium trichloride GTF, the glucose metabolism of the cells was significantly improved, and the optimum concentration was 1.0 渭 g ml ~ (-1). The model of insulin resistance in vitro was established by HepG2 cells. The method is simple, reliable and reproducible, and can be widely used in the study of the hypoglycemic function of natural active substances.
【作者单位】：中国农业科学院农产品加工研究所;天津科技大学食品工程与生物技术学院;
【基金】：国家自然基金面上项目(31371763) 中央级公益性科研院所基本科研业务费专项(S2016JC01)
【分类号】：R91

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