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巨噬细胞条件培养基对胰岛素调节骨骼肌细胞作用的影响

刘婕1,牛文彦1,2   

  1. 1. 天津医科大学
    2.
  • 收稿日期:2010-09-30 修回日期:2010-11-17 出版日期:2011-02-15 发布日期:2011-02-15
  • 通讯作者: 刘婕

The Effect of the Conditioned Medium from Macrophages on Insulin Function in Skeletal Muscle Cells

  • Received:2010-09-30 Revised:2010-11-17 Published:2011-02-15 Online:2011-02-15
  • Contact: Jie LIU

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2009

摘要： 摘要 目的：探讨常氧和缺氧培养的巨噬细胞条件培养基对胰岛素调节骨骼肌葡萄糖转运子4（GLUT4）作用的影响。方法：常氧和缺氧培养巨噬细胞，提取条件培养基孵育C2C12GLUT4myc骨骼肌细胞，用偶联抗体的吸光度法测定细胞膜上GLUT4myc的含量，Real-time PCR法测定巨噬细胞TNF-α mRNA的表达，ELISA法测定巨噬细胞条件培基中TNF-α的含量。结果：常氧和缺氧培养的巨噬细胞条件培养基削弱胰岛素刺激的骨骼肌细胞GLUT4myc转位(P<0.01)；缺氧培养的巨噬细胞TNF-α mRNA和蛋白表达均显著高于常氧培养的巨噬细胞（P<0.05），但两种条件培养基对骨骼肌中胰岛素作用的影响无显著性差异。结论：常氧培养和缺氧培养的巨噬细胞条件培养基均直接造成骨骼肌细胞胰岛素抵抗，缺氧的巨噬细胞条件培养基并不能加重骨骼肌胰岛素抵抗。

关键词: 巨噬细胞, 缺氧, 条件培基, C2C12GLUT4myc, 转位

Abstract: Abstract Objective: To explore the effect of conditioned media (CM) from nomoxia- and hypoxia-treated macrophages on insulin-regulated glucose transporter 4 (GLUT4) in skeletal muscle cells. Methods: The macrophages were cultured under normoxia or hypoxia condition, respectively. Then conditioned media were collected and used to incubate C2C12GLUT4myc skeletal muscle cells. The levels of GLUT4myc on the cell surface were measured by an antibody-coupled absorbance assay. The mRNA expression of TNF-α in macrophages was determined by real-time PCR. The levels of TNF-α in the media of macrophages were determined by ELISA. Results: The increase of insulin-stimulated GLUT4myc translocation was significantly impaired under incubation with CM from both nomoxia- and hypoxia-treated macrophages (P<0.01). Both mRNA and protein expression of TNF-α were significantly higher in CM from hypoxia-treated macrophages than nomoxia-treated ones (P<0.05). But there was no difference between two CMs on insulin function in skeletal muscle cells. Conclusion: Both CM from nomoxia- and hypoxia-treated macrophages directly induce insulin resistance in skeletal muscle cells. CM from hypoxia-treated macrophages does not worsen insulin resistance.

Key words: Macrophages, Hypoxia, Conditioned medium, C2C12GLUT4myc, Translocation.