Axin1调节骨骼肌GLUT4蛋白表达的作用研究

目的：探讨体轴发育抑制因子（Axin1）调节骨骼肌葡萄糖转运蛋白4(GLUT4)蛋白表达的作用及其机制。方法：利用RNAi干扰的Axin1腺病毒（Ad-siAxin1）感染C2C12小鼠骨骼肌细胞敲低Axin1，荧光显微镜和MTS实验明确Ad-siAxin1最佳感染浓度和时间。分别用携带绿色荧光蛋白的腺病毒（Ad-GFP）、Ad-siAxin1、载体质粒（vector）和Axin1质粒转染C2C12小鼠骨骼肌细胞敲低或过表达Axin1蛋白，Western印迹检测Axin1、端锚聚合酶蛋白（TNKS）和GLUT4蛋白水平。结果：荧光显微镜和MTS实验结果显示，在C2C12小鼠骨骼肌细胞中Ad-siAxin1作用的最佳浓度和时间分别为160μL和48 h。Western印迹结果显示，与Ad-GFP组相比，Ad-siAxin1组中Axin1蛋白降低（t=6.746,P<0.01）。Ad-siAxin1组TNKS蛋白水平降低（t=4.019,P<0.05）,GLUT4蛋白水平下调（t=3.248,P<0.05）。与vector组相比，转染Axin1质粒后，Axin1蛋白水平上...

Study the effect of Axin1 on regulating GLUT4 protein in skeletal muscle

Objective: To investigate the role and molecular mechanism of Axin formation inhibitor（Axis1） in regulating glucose transporter 4（GLUT4） protein in skeletal muscle cells. Methods:C2C12 mouse skeletal muscle cells were infected with adenovirus expressing siAxin1（Ad-siAxin1） to knock down Axin1. Fluorescence microscopy and MTS experiments were measured to clarify the optimal infection concentration and time of Ad-siAxin1. C2C12 cells were infected with Ad-GFP（adenovirus expressing green fluorescent protein） or Ad-siAxin1 to knock down Axin1，and transfected with vector plasmid or Axin1 plasmid to overexpress Axin1. The expression of Axin1，Tankyrase（TNKS） and GLUT4 proteins were detected by Western blotting. Results:The results of fluorescence microscopy and MTS experiments showed that the optimal concentration and time of Ad-siAxin1 in C2C12 skeletal muscle cells were 160 μL and 48 h. Western blotting results showed that compared with the Ad-GFP group，Axin1 protein in Ad-siAxin1 group was decreased（t=6.746，P<0.01）. Ad-siAxin1 significantly decreased TNKS protein（t=4.019，P<0.05），and down-regulated GLUT4 protein（t=3.248，P<0.05）.The expression of Axin1 protein was up-regulated after transfection with Axin1 plasmid（t=4.868，P<0.01）.The protein of TNKS and GLUT4 were significantly up-regulated（t=4.897，4.789，both P<0.01）. Conclusion:Axin1 may regulate the level of GLUT4 protein through TNKS.