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胰岛素激活PI3K-AKT通路而促进H9c2心肌细胞葡萄糖摄取

, PP. 538-542

Keywords: 胰岛素,-NBDG,Hc心肌细胞,葡萄糖摄取,PIK-AKT,pMAPK,GLUT和GLUT

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Abstract:

目的探索胰岛素(Insulin)对H9c2心肌细胞葡萄糖摄取的影响及其与PI3K-AKT信号通路的关系。方法采用不同时间及不同浓度的胰岛素处理H9c2心肌细胞,确定胰岛素处理H9c2细胞的最佳浓度和最适时间。实验分空白对照组(NC组),2-NBDG组,Insulin组,Insulin+PI3K-AKT通路抑制剂LY294002组(LY294002组),Insulin+p38MAPK通路抑制剂BIRB796组(BIRB796组),使用荧光标记2-脱氧葡萄糖(2-NBDG)检测H9c2心肌细胞葡萄糖摄取能力,以流式细胞仪和荧光酶标仪检测心肌细胞对葡萄糖摄取的变化。通过间接免疫荧光法及激光共聚焦检测H9c2心肌细胞葡萄糖转运体-1(glucosetansporter-1,GLUT-1)及葡萄糖转运体-4(glucosetansporter-4,GLUT-4)的表达。结果①2-NBDG可用于检测H9c2心肌细胞葡萄糖摄取能力。②Insulin处理H9c2心肌细胞最适浓度和时间分别为200μmol/L和30min。③Insulin组与对照组相比增加H9c2心肌细胞葡萄糖摄取(P<0.05);LY294002组与Insulin组相比降低心肌细胞葡萄糖摄取,两组间差异明显(P<0.05),BIRB796组较Insulin组无统计学差异(P>0.05),提示LY294002可抑制Insulin促进心肌细胞葡萄糖摄取,而BIRB796不能抑制上述作用。④H9c2心肌细胞膜表达葡萄糖转运体1(GLUT1)和葡萄糖转运体4(GLUT4),Insulin处理H9c2心肌细胞30min后经激光共聚焦成像显示H9c2心肌细胞膜GLUT1和GLUT4表达增加(P<0.05)。结论胰岛素促进H9c2心肌细胞葡萄糖摄取的作用可能与PI3K-AKT通路相关,与p38MAPK通路不相关,胰岛素激活PI3K-AKT信号通路后可能促进H9c2心肌细胞膜上GLUT-1及GLUT-4的表达,从而使H9c2心肌细胞对葡萄糖摄取增加。

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