缺氧诱导因子-1α过表达通过葡萄糖转运载体-4介导缺氧大鼠骨髓间充质干细胞对葡萄糖的摄取

HIF-1αgene improves glucose intake of hypoxic bone marrow mesenchymal stem cells by GLUT-4 ex vivo

目的探列缺氧诱导因子-1α（HIF-1α）基因转染是否提高骨髓间充质干细胞（MSC）缺氧状态下的葡萄糖摄取能力以及这种能力是否与葡萄糖转运载体-4（GLUT-4）的表达和易位有关。方法将MSCs分为常氧非转染组（即对照组）、常氧转染组、缺氧非转染组和缺氧转染组，分别于常氧（5％CO2，37℃）和缺氧（94％N2、1％0z和5％CO2，37℃）条件下孵育8h。放射同位素法检测。H—G摄取量，免疫细胞化学和Western-blot检测GLUT-4的表达。结果①与缺氧非转染组相比，缺氧转染组显著提高细胞摄取。H—G量（P〈0．01），但二者均显著低于对照组和常氧转染组对。H—G的摄取（P〈0．01）；②与缺氧非转染组相比，缺氧转染组显著提高细胞和细胞膜GLUT-4蛋白的表达（P〈0．01），2组均显著低于对照组GLUT-4蛋白的表达和移位（P〈0．01）；③。H—G摄取量与细胞膜中GLUT-4表达呈正相关（r=0．437，P=0．001）。结论HIF-1a基因提高MSCs的葡萄糖摄取能力，此机制与HIF-1α基因提高GLUT-4的表达和易位相关。

Objective To elucidate whether hypoxia induced factor-1α（HIF-1α） gene improves the capability of glucose intake in hypoxia bone mesenchymal stem cells （MSCs）, and whether the transaction and the expression of GLUT-4 play an important role in this process ex vivo . Methods MSCs were randomly divided into control （normoxia non-HIF-1α transfection）, normoxia HIF-1α transfection group, hypoxia non-HIF-1α transfection group, hypoxia Hif-la transfection group, and then each group was cultured with normoxia （5%CO2 at 37 ℃ ） or hypoxia（94%N2, 1%O2, 5%CO2 at 37 ℃） for 8 h. Finally, glucose uptake and the expression of GLUT-4 in cells and in cell membrane were assayed. Results （1）3 H-G intake of MSCs was significantly increased in HIF-la gene transfection than in non-HIF-1α gene transfection （ P 〈0.01） when hypoxia, however, those were significantly lower in each hypoxla group than in each normoxia group （ P〈0.01） ; （2）Under hypoxia, HIF-la significantly increased the expression of GLUT-4 protein in cells and in cell membranes （ P 〈0. 01）, which were significantly decreased than in control group and in normoxia HIF-1α transfection group （P〈0.01） ; （3）There was significant positive relation between 3 H-G intake and GLUT-4 protein in plasma membrane （ r = 0. 437, P = 0. 001）. Conclusion HIF-1α gene significantly improved the capability of glucose intake of hypoxia MSCs, which was mediated by the expression and transaction of GLUT-4.