Angiopoietin-Related Growth Factor Enhances Blood Flow Via Activation of the ERK1/2-eNOS-NO Pathway in a Mouse Hind-Limb Ischemia Model
Angiopoietin-Related Growth Factor Enhances Blood Flow Via Activation of the ERK1/2-eNOS-NO Pathway in a Mouse Hind-Limb Ischemia Model
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DOI：
10.1161/atvbaha.107.149674
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发表时间：
2008-05
期刊：
影响因子：
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通讯作者：
Takashi Urano;Yasuhiro Ito;M. Akao;T. Sawa;K. Miyata;M. Tabata;T. Morisada;T. Hato;M. Yano
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文献类型：
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作者：
Takashi Urano;Yasuhiro Ito;M. Akao;T. Sawa;K. Miyata;M. Tabata;T. Morisada;T. Hato;M. Yano
目的：过量表达血管生成素相关生长因子(AGF)的转基因小鼠表现出促进血管生成的作用，提示AGF可能是治疗缺血性疾病的有用药物靶点。我们的目标是确定AGF是否在小鼠后肢缺血模型中增强血流，并确定AGF信号转导内皮细胞的分子机制。方法和结果：肌肉注射携带AGF的腺病毒到缺血肢体可增加AGF的产生，从而通过诱导血管生成和动脉生成增加血流量，从而减少截肢的必要性。体外分析表明，人脐静脉内皮细胞暴露于AGF可通过激活ERK1/2-内皮型一氧化氮合酶(ENOS)信号通路增加一氧化氮(NO)的产生。AGF刺激的eNOS磷酸化、NO产生和内皮细胞迁移均被特定的MEK1/2抑制剂阻断。此外，无论是接受一氧化氮合酶抑制剂L-NAME的小鼠，还是eNOS基因敲除的小鼠，血管生长因子都不能恢复缺血后肢的血流。结论：激活ERK1/2-eNOS-NO通路是AGF通过诱导血管生成和动脉生成增加血流量的重要信号机制。进一步研究AGF信号通路的调控机制可能有助于开发治疗缺血性血管疾病的新的临床策略。
Objective—Transgenic mice overexpressing angiopoietin-related growth factor (AGF) exhibit enhanced angiogenesis, suggesting that AGF may be a useful drug target in ischemic disease. Our goal was to determine whether AGF enhances blood flow in a mouse hind-limb ischemia model and to define molecular mechanisms underlying AGF signaling in endothelial cells. Methods and Results—Intramuscular injection of adenovirus harboring AGF into the ischemic limb increased AGF production, which increased blood flow through induction of angiogenesis and arteriogenesis, thereby reducing the necessity for limb amputation. In vitro analysis showed that exposing human umbilical venous endothelial cells to AGF increased nitric oxide (NO) production through activation of an ERK1/2-endothelial NO synthetase (eNOS) signaling pathway. AGF-stimulated eNOS phosphorylation, NO production, and endothelial cell migration were all abolished by specific MEK1/2 inhibitors. Moreover, AGF did not restore blood flow to ischemic hind-limbs of either mice receiving NOS inhibitor L-NAME or eNOS knockout mice. Conclusion—Activation of an ERK1/2-eNOS-NO pathway is a crucial signaling mechanism by which AGF increases blood flow through induction of angiogenesis and arteriogenesis. Further investigation of the regulation underlying AGF signaling pathway may contribute to develop a new clinical strategy for ischemic vascular diseases.