通过细胞实验探讨大豆肽与豌豆肽的复合物对II型糖尿病胰岛素抵抗作用,并初步探明其作用机制。采用1×10-6 mol/L的人胰岛素处理HepG2细胞24 h建立胰岛素抵抗模型;CCK-8法确定大豆肽及豌豆肽对HepG2细胞的安全剂量;检测不同复配比例的大豆肽和豌豆肽实验组的葡萄糖消耗、葡萄糖吸收及细胞中的糖原含量反应糖代谢情况,通过Western Blot法检测各蛋白的表达。结果表明,与对照组相比,HepG2细胞置于含1×10-6 mol/L人胰岛素培养液孵育24 h,葡萄糖消耗量、葡萄糖吸收量、糖原含量都降低,表明建模成功。大豆肽及豌豆肽干预均能改善HepG2细胞的胰岛素抵抗状况,提高细胞对葡萄糖的吸收和消耗能力、糖原合成能力,并能够提高葡萄糖转运蛋白GLUT2、GLUT4的表达量及胰岛素受体底物-1(insulin receptor substrate-1,IRS-1)和蛋白激酶B(Akt)蛋白的磷酸化水平,其中质量浓度为200 μg/mL、比例为2∶1的大豆肽和豌豆肽效果最好。大豆肽与豌豆肽以2∶1的质量比进行复配可以有效改善HepG2细胞的胰岛素抵抗状况,可能与肽提高细胞中GLUT2和GLUT4的表达量,以及提高IRS-1和Akt蛋白的磷酸化水平有关。
This study aimed to investigate the effects of soybean oligopeptides and pea oligopeptides on insulin resistance in type 2 diabetes mellitus by cell experiment and preliminarily explore the mechanism.The insulin resistance model was established by HepG2 cells treated with 1×10-6 mol/L human insulin for 24 h.The safe dose of soybean oligopeptides and pea oligopeptides to HepG2 cells was determined by the CCK-8 method.The glucose consumption, absorption, and glycogen content in cells were measured in the experimental groups with different proportions of soybean oligopeptides and pea oligopeptides.The expressions of IRS-1, p-IRS-1, Akt, p-Akt, GLUT2, and GLUT4 were detected by Western Blot.Results showed that compared with the control group, HepG2 cells were incubated in human insulin medium containing 1×10-6 mol/L for 24 h, and glucose consumption, glucose absorption and glycogen content were all reduced, indicating successful modelling.Both soybean oligopeptides and pea oligopeptides could improve the insulin resistance of HepG2 cells and have effective improvements, including glucose absorption and consumption ability and glycogen synthesis ability.The peptides could increase the expression of glucose transporters GLUT2 and GLUT4 and the phosphorylation levels of IRS-1 and Akt.And the most effective concentration was 200μg/mL and the ratio of soybean oligopeptides to pea oligopeptides was 2∶1.The 2∶1 combination of soybean oligopeptides and pea oligopeptides can effectively improve the insulin resistance of HepG2 cells, which may be related to the increased expression of glucose transporters GLUT2 and GLUT4 and the increased phosphorylation of IRS-1 and Akt.
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