Superoxide induced by a high-glucose concentration attenuates production of angiogenic growth factors in hypoxic mouse mesenchymal stem cells
Superoxide induced by a high-glucose concentration attenuates production of angiogenic growth factors in hypoxic mouse mesenchymal stem cells
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DOI：
10.1677/joe-10-0305
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发表时间：
2011-02-01
影响因子：
4
通讯作者：
Watanabe, Yasuhiro
中科院分区：
文献类型：
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作者：
Ishizuka, Toshiaki;Hinata, Takashi;Watanabe, Yasuhiro
以往的研究表明，旁分泌系统可能是骨髓间充质干细胞（MSC）治疗缺血性疾病的重要介质。高血压和缺氧与活性氧水平增加有关;因此，氧化应激可能会影响糖尿病患者在缺氧条件下MSC的旁分泌作用，尽管这种作用的机制尚不清楚。低氧诱导因子1 α（HIF-1 α）调节低氧诱导基因的转录。我们确定了高浓度葡萄糖对缺氧间充质干细胞通过HIF-1 α诱导产生血管生成生长因子的影响。MSC用不同的葡萄糖浓度（5.6、11、20或30 mM）培养24 h。然后将细胞在低氧室（5%O-2）或常氧（21%O-2）下孵育6或24 h。葡萄糖以剂量依赖性方式减弱缺氧MSC中HIF-1 α、血管内皮生长因子A（165）（VEGF-A（165））和血小板衍生生长因子B（PDGF-B）的蛋白水平。用蛋白酶体活性的特异性抑制剂MG 132处理，显著逆转了低氧MSC中高糖浓度的抑制作用。4-羟基-四甲基哌啶-氧基（一种细胞可渗透的超氧化物清除剂）或夹竹桃素（一种NADPH氧化酶抑制剂）显著逆转葡萄糖诱导的VEGF-A（165）、PDGF-B和HIF-1 α蛋白水平的衰减。高浓度葡萄糖（30 mM）的刺激显着增加细胞内的超氧化物水平在缺氧的MSC。我们的研究结果表明，在缺氧MSC中，高浓度葡萄糖诱导的细胞内超氧化物水平的增加可能会减弱缺氧诱导的HIF-1 α表达，这反过来又减弱了缺氧诱导的VEGF-A（165）和PDGF-B转录。内分泌学杂志（2011）208，147-159
Previous reports have shown that the paracrine system may be an important mediator in bone-marrow-derived mesenchymal stem cell (MSC) therapy for ischemic diseases. Hyperglycemia and hypoxia have been associated with increased levels of reactive oxygen species; oxidative stress may therefore influence the paracrine effects of MSCs under hypoxic conditions in diabetic patients, although the mechanism underlying this effect remains unknown. Hypoxia-inducible factor 1 alpha (HIF-1 alpha) regulates the transcription of hypoxia-inducible genes. We determined the effect of high-glucose concentrations on the production of angiogenic growth factors via HIF-1 alpha induction in hypoxic MSCs. MSCs were cultured with different glucose concentration (5.6, 11, 20, or 30 mM) for 24 h. The cells were then incubated in a hypoxic chamber (5% O-2) or under normoxia (21% O-2) for 6 or 24 h. Protein levels of HIF-1 alpha, vascular endothelial growth factor A(165) (VEGF-A(165)), and platelet-derived growth factor B (PDGF-B) were attenuated by glucose in hypoxic MSCs in a dose-dependent manner. Treatment with MG132, a specific inhibitor of proteasome activity, significantly reversed the inhibitory effect of high-glucose concentrations in hypoxic MSCs. 4-Hydroxyl-tetramethylpiperidin-oxyl (a cell-permeable superoxide scavenger) or Apocynin (a NADPH oxidase inhibitor) significantly reversed glucose-induced attenuation of VEGF-A(165), PDGF-B, and HIF-1 alpha protein levels. Stimulation with a high-glucose concentration (30 mM) significantly increased intracellular superoxide levels in hypoxic MSCs. Our results suggest that in hypoxic MSCs the increase in intracellular superoxide levels induced by high-glucose concentrations may attenuate hypoxia-induced HIF-1 alpha expression, which in turn attenuates hypoxia-induced VEGF-A(165) and PDGF-B transcription. Journal of Endocrinology (2011) 208, 147-159