[1]叶林,陆凯,张冬颖,等.胰岛素激活PI3K-AKT通路而促进H9c2心肌细胞葡萄糖摄取[J].陆军军医大学学报(原第三军医大学学报),2015,37(06):538-542.
 Ye Lin,Lu Kai,Zhang Dongying,et al.Insulin promotes glucose uptake via PI3K-AKT pathway in H9c2 myocardial cells[J].J Amry Med Univ (J Third Mil Med Univ),2015,37(06):538-542.
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陆军军医大学学报(原第三军医大学学报)[ISSN:1000-5404/CN:51-1095/R]

卷:
37卷
期数:
2015年第06期
页码:
538-542
栏目:
论著
出版日期:
2015-03-30

文章信息/Info

Title:
Insulin promotes glucose uptake via PI3K-AKT pathway in H9c2 myocardial cells
作者:
叶林陆凯张冬颖覃数
重庆医科大学附属第一医院:重症医学科,心血管内科
Author(s):
Ye Lin Lu Kai Zhang Dongying Qin Shu

Department of Critical Care Medicine, Department of Cardiovascular Medicine, the First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016; China

关键词:
胰岛素2-NBDGH9c2心肌细胞葡萄糖摄取PI3K-AKTp38MAPKGLUT1和GLUT4
Keywords:
insulin H9c2 cells 2-NBDG glucose uptake PI3K-AKT p38MAPK GLUT1 GLUT4
分类号:
Q578;R322.11;R329.26
文献标志码:
A
摘要:

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(glucose tansporter-1,GLUT-1)及葡萄糖转运体-4(glucose tansporter-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200 μmol/L和30 min。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。

Abstract:

Objective      To explore the effect of insulin on glucose uptake in H9c2 myocardial cells and its relationship with PI3K-AKT pathway.       Method      A fluorescence microplate reader was used to determine the optimal concentration and incubation time of insulin for H9c2 cells. The H9c2 cells were divided to 5 groups: a normal control group, a 2-[N-(7-nitrobenz-2-oxa-1,3-diaxol-4-yl)amino]-2-deoxyglucose (2-NBDG) group, an insulin group, an insulin+LY294002 (PI3K-AKT pathway inhibitor) group, and an insulin+BIRB796 (p38MAPK pathway inhibitor) group. The glucose uptake was analyzed with the fluorescence microplate reader and a flow cytometer. The expression of glucose tansporter-1 (GLUT1) and glucose tansporter-4 (GLUT4) in the H9c2 cells was detected by indirect immunofluorescence assay and confocal laser scanning analysis.       Results      2-NBDG could be used to detect glucose uptake in H9c2 cells. The optimal concentration and incubation time of insulin were 200 μmol/L and 30 min, respectively. Insulin could promote glucose uptake in the H9c2 cells (P<0.05), and this effect was blocked by LY294002 (P<0.05) but not by BIRB796 (P>0.05). GLUT1 and GLUT4 were expressed in the H9c2 cell membranes, and insulin increased the expressions of GLUT1and GLUT4.       Conclusion      Insulin can increase the expression of GLUT1and GLUT4 via PI3K-AKT pathway rather than p38MAPK pathway, and then promote glucose uptake in the H9c2 cells.

相似文献/References:

[1]叶林,陆凯,张冬颖,等.2-NBDG检测H9c2心肌细胞葡萄糖摄取能力的研究[J].陆军军医大学学报(原第三军医大学学报),2012,34(15):1533.
 Ye Lin,Lu Kai,Zhang Dongying,et al.2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino]-2-deoxyglucose as fluorescence probe to detect glucose uptake in H9c2 cells[J].J Amry Med Univ (J Third Mil Med Univ),2012,34(06):1533.

更新日期/Last Update: 2015-03-20