骨骼肌细胞去极化和收缩调节GLUT4转位的机制研究

发布时间：2018-02-14 03:15

  本文关键词： 葡萄糖转运子4 去极化 收缩 转位　出处：《天津医科大学》2009年硕士论文　论文类型：学位论文

【摘要】：目的： 本研究应用稳定表达GLUT4myc的L6GLUT4myc和C2C12GLUT4myc两种骨骼肌细胞模型探讨去极化和收缩调节GLUT4myc转位的机制。 方法： 第一部分比较55mM高钾溶液诱发的L6GLUT4myc肌管去极化和C2C12GLUT4myc肌管收缩对GLUT4myc转位的影响。测定两种细胞GLUT4myc转位响应高钾作用的时间曲线。第二部分比较55mM高钾溶液刺激L6GLUT4myc肌管和C2C12GLUT4myc肌管GLUT4myc转位过程中的信号分子的作用,检测信号分子的磷酸化。在C2C12GLUT4myc肌管中通过使用不同蛋白激酶抑制剂,分析高钾刺激GLUT4myc转位的信号机制。 结果： L6GLUT4myc肌管的GLUT4myc转位对高钾诱发的去极化作用呈时间依赖关系,最大值为基础状态的1.67±0.41倍(P0.05)：C2C12GLUT4myc肌管的GLUT4myc转位对高钾诱发的收缩作用同样呈时间依赖关系,最大值为基础状态的1.51±0.15倍(P0.05)。 比较高钾诱导的去极化和收缩对信号分子的作用结果显示,在L6GLUT4myc肌管中去极化刺激增强AMPK和CaMKⅡ以及AS160的磷酸化；在C2C12GLUT4myc肌管中收缩同样刺激增强AMPK和CaMKⅡ以及AS160的磷酸化。在C2C12GLUT4myc肌管中,骨骼肌肌球蛋白ATPase抑制剂BTS可抑制58%(P0.05)的GLUT4myc的转位,AMPK的抑制剂Compound C可抑制49%(P0.05)的GLUT4myc的转位,钙离子螯合剂BAPTA-AM可抑制42%(P0.05)的GLUT4myc的转位,CaMKⅡ抑制剂KN93可抑制50%(P0.05)的GLUT4myc的转位。 结论： 1、不可收缩的L6GLUT4myc肌管和可收缩的C2C12GLUT4myc肌管的GLUT4myc转位对高钾作用呈时间依赖关系,去极化和收缩均可使细胞膜GLUT4myc增加。 2、高钾溶液诱发的去极化和收缩刺激两种肌管GLUT4myc转位,信号分子AMPK、CaMKⅡ和AS160的磷酸化都增强,这三种信号分子都响应去极化和收缩作用。 3、收缩刺激C2C12GLUT4myc肌管膜GLUT4myc增加时,收缩、Ca2+及信号蛋白AMPK和CaMKⅡ参与GLUT4myc的转位。
[Abstract]:Objective:. In this study, two skeletal muscle cell models, L6GLUT4myc and C2C12GLUT4myc, which stably expressed GLUT4myc, were used to investigate the mechanism of depolarization and contraction regulating GLUT4myc translocation. Methods:. In the first part, we compare the effects of depolarization of L6GLUT4myc muscle tube induced by 55mm high potassium solution and contraction of C2C12GLUT4myc myc muscle tube on GLUT4myc transposition. The time curves of GLUT4myc transposition in two kinds of cells in response to hyperkalemia were measured. The second part compared the effects of 55 mm high potassium solution on L6GLUT4myc. The role of signaling molecules in the GLUT4myc translocation of muscle tubes and C2C12GLUT4myc muscle tubes. The signal mechanism of GLUT4myc translocation stimulated by high potassium was analyzed by using different protein kinase inhibitors in C2C12GLUT4myc myc muscle tube. Results:. The GLUT4myc transposition of L6GLUT4myc muscle tube showed a time-dependent effect on the depolarization induced by hyperkalemia. The maximum value was 1.67 卤0.41 times of the base state (P0.05: C2C12GLUT4myc myc myc muscle tube) and the contraction induced by hyperkalemia was also time-dependent. The maximum value was 1.51 卤0.15 times of the base state (P0.05). The effects of depolarization and contraction induced by high potassium on signal molecules were compared. The results showed that the phosphorylation of AMPK, CaMK 鈪，

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