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目的研究明日叶查尔酮(Angelica keiskei chalcones,AC)对胰岛素抵抗L6细胞磷脂酰肌醇3激酶-蛋白激酶-葡萄糖转运体4(PI3K-Akt-GLUT4)信号通路的影响。方法采用高浓度胰岛素诱导的胰岛素抵抗L6细胞,分别加入5、10、20μmol/L终浓度的明日叶查尔酮共同培养24 h,葡萄糖氧化酶法测定细胞上清液中葡萄糖含量,逆转录聚合酶链式反应方法检测细胞PI3K和Akt的mRNA表达水平,蛋白印迹法检测GLUT4和Akt的蛋白表达水平。结果胰岛素抵抗组细胞培养液的葡萄糖含量显著高于空白对照组(P<0.05),而PI3K mRNA、Akt mRNA和GLUT4、Akt蛋白表达水平则降低(P<0.05);中、高剂量AC组细胞培养液葡萄糖含量均低于胰岛素抵抗组(P<0.05),而PI3K mRNA、Akt mRNA和GLUT4、Akt蛋白表达水平则升高(P<0.05)。结论明日叶查尔酮可上调L6细胞胰岛素抵抗模型PI3K-AktGLUT4信号通路的表达水平,增强其对葡萄糖的摄取利用,改善胰岛素抵抗。
Objective To investigate the effect of ACEI on PI3K-Akt-GLUT4 signaling pathway in insulin-resistant L6 cells. Methods Insulin-resistant L6 cells were induced by high concentration of insulin. Melatonin (5, 10, and 20μmol / L) were added into the culture for 24 h. Glucose oxidase was used to determine the glucose content in the supernatant. Reverse transcription polymerase chain reaction The mRNA expression of PI3K and Akt was detected by enzyme-linked immunosorbent assay. The protein expression of GLUT4 and Akt was detected by Western blotting. Results The glucose content of cell culture medium of insulin resistance group was significantly higher than that of blank control group (P <0.05), while the expression of PI3K mRNA, Akt mRNA and GLUT4 and Akt protein were decreased (P <0.05) The content of glucose in culture medium was lower than that in insulin resistance group (P <0.05), while the expression of PI3K mRNA, Akt mRNA and GLUT4, Akt protein increased (P <0.05). Conclusion Yechal ketone could up-regulate the expression of PI3K-AktGLUT4 signaling pathway in insulin resistance model L6 cells and enhance its uptake and utilization of glucose to improve insulin resistance.