本文關(guān)鍵詞： 葡萄糖轉(zhuǎn)運(yùn)子4 去極化 收縮 轉(zhuǎn)位　出處：《天津醫(yī)科大學(xué)》2009年碩士論文　論文類型：學(xué)位論文

【摘要】：目的： 本研究應(yīng)用穩(wěn)定表達(dá)GLUT4myc的L6GLUT4myc和C2C12GLUT4myc兩種骨骼肌細(xì)胞模型探討去極化和收縮調(diào)節(jié)GLUT4myc轉(zhuǎn)位的機(jī)制。 方法： 第一部分比較55mM高鉀溶液誘發(fā)的L6GLUT4myc肌管去極化和C2C12GLUT4myc肌管收縮對(duì)GLUT4myc轉(zhuǎn)位的影響。測定兩種細(xì)胞GLUT4myc轉(zhuǎn)位響應(yīng)高鉀作用的時(shí)間曲線。第二部分比較55mM高鉀溶液刺激L6GLUT4myc肌管和C2C12GLUT4myc肌管GLUT4myc轉(zhuǎn)位過程中的信號(hào)分子的作用,檢測信號(hào)分子的磷酸化。在C2C12GLUT4myc肌管中通過使用不同蛋白激酶抑制劑,分析高鉀刺激GLUT4myc轉(zhuǎn)位的信號(hào)機(jī)制。 結(jié)果： L6GLUT4myc肌管的GLUT4myc轉(zhuǎn)位對(duì)高鉀誘發(fā)的去極化作用呈時(shí)間依賴關(guān)系,最大值為基礎(chǔ)狀態(tài)的1.67±0.41倍(P0.05)：C2C12GLUT4myc肌管的GLUT4myc轉(zhuǎn)位對(duì)高鉀誘發(fā)的收縮作用同樣呈時(shí)間依賴關(guān)系,最大值為基礎(chǔ)狀態(tài)的1.51±0.15倍(P0.05)。 比較高鉀誘導(dǎo)的去極化和收縮對(duì)信號(hào)分子的作用結(jié)果顯示,在L6GLUT4myc肌管中去極化刺激增強(qiáng)AMPK和CaMKⅡ以及AS160的磷酸化；在C2C12GLUT4myc肌管中收縮同樣刺激增強(qiáng)AMPK和CaMKⅡ以及AS160的磷酸化。在C2C12GLUT4myc肌管中,骨骼肌肌球蛋白ATPase抑制劑BTS可抑制58%(P0.05)的GLUT4myc的轉(zhuǎn)位,AMPK的抑制劑Compound C可抑制49%(P0.05)的GLUT4myc的轉(zhuǎn)位,鈣離子螯合劑BAPTA-AM可抑制42%(P0.05)的GLUT4myc的轉(zhuǎn)位,CaMKⅡ抑制劑KN93可抑制50%(P0.05)的GLUT4myc的轉(zhuǎn)位。 結(jié)論： 1、不可收縮的L6GLUT4myc肌管和可收縮的C2C12GLUT4myc肌管的GLUT4myc轉(zhuǎn)位對(duì)高鉀作用呈時(shí)間依賴關(guān)系,去極化和收縮均可使細(xì)胞膜GLUT4myc增加。 2、高鉀溶液誘發(fā)的去極化和收縮刺激兩種肌管GLUT4myc轉(zhuǎn)位,信號(hào)分子AMPK、CaMKⅡ和AS160的磷酸化都增強(qiáng),這三種信號(hào)分子都響應(yīng)去極化和收縮作用。 3、收縮刺激C2C12GLUT4myc肌管膜GLUT4myc增加時(shí),收縮、Ca2+及信號(hào)蛋白AMPK和CaMKⅡ參與GLUT4myc的轉(zhuǎn)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 鹵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 鹵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，

本文編號(hào)：1509748