Direct Activation of NADPH Oxidase 2 by 2-Deoxyribose-1-Phosphate Triggers Nuclear Factor Kappa B-Dependent Angiogenesis.
Direct Activation of NADPH Oxidase 2 by 2-Deoxyribose-1-Phosphate Triggers Nuclear Factor Kappa B-Dependent Angiogenesis.
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DOI：
10.1089/ars.2016.6869
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发表时间：
2018-01-10
影响因子：
6.6
通讯作者：
Pula G
中科院分区：
文献类型：
--
作者：
Vara D;Watt JM;Fortunato TM;Mellor H;Burgess M;Wicks K;Mace K;Reeksting S;Lubben A;Wheeler-Jones CPD;Pula G
目的：脱氧核糖-1-磷酸(DRP)是一种促血管生成旁分泌刺激物质，由癌细胞、血小板和巨噬细胞释放，作用于内皮细胞。这项研究的目的是阐明DRP如何刺激人内皮细胞的血管生成反应。结果：活细胞成像、电子顺磁共振、DRP相互作用蛋白的下拉、免疫印迹、不同NADPH氧化酶(NOXs)的基因沉默、小干扰RNA(SiRNA)及其调节辅基转移以及糖转运体葡萄糖转运体1(GLUT1)抑制剂的实验表明，DRP通过直接激活内皮细胞NOX2复合体而不是NOX4发挥作用。NOX2活性引起的活性氧生成增加导致转录因子核因子kappaB(NF-κB)氧化还原依赖的激活，进而诱导血管内皮生长因子受体2上调。通过血管内皮细胞形成实验、小干扰RNA基因沉默和基于抗体的受体抑制，我们证明了核因子-κB和血管内皮生长因子受体2的激活是血管生成反应所必需的。在−/−小鼠体内的切除创伤和Matrigel Plug血管形成实验中，证实了DRP上调VEGFR2和依赖于NOX2的血管生成。创新：我们首次证明DRP在细胞内发挥作用，并通过直接结合和激活NOX2酶复合体刺激超氧阴离子的产生。结论：本研究揭示了DRP促血管生成活性的一种新的分子机制，该机制涉及NOX2和NF-κB的顺序激活以及VEGFR2的上调。抗氧化剂。氧化还原信号。28,110-130。
Aims: Deoxyribose-1-phosphate (dRP) is a proangiogenic paracrine stimulus released by cancer cells, platelets, and macrophages and acting on endothelial cells. The objective of this study was to clarify how dRP stimulates angiogenic responses in human endothelial cells. Results: Live cell imaging, electron paramagnetic resonance, pull-down of dRP-interacting proteins, followed by immunoblotting, gene silencing of different NADPH oxidases (NOXs), and their regulatory cosubunits by small interfering RNA (siRNA) transfection, and experiments with inhibitors of the sugar transporter glucose transporter 1 (GLUT1) were utilized to demonstrate that dRP acts intracellularly by directly activating the endothelial NOX2 complex, but not NOX4. Increased reactive oxygen species generation in response to NOX2 activity leads to redox-dependent activation of the transcription factor nuclear factor kappa B (NF-κB), which, in turn, induces vascular endothelial growth factor receptor 2 (VEGFR2) upregulation. Using endothelial tube formation assays, gene silencing by siRNA, and antibody-based receptor inhibition, we demonstrate that the activation of NF-κB and VEGFR2 is necessary for the angiogenic responses elicited by dRP. The upregulation of VEGFR2 and NOX2-dependent stimulation of angiogenesis by dRP were confirmed in excisional wound and Matrigel plug vascularization assays in vivo using NOX2−/− mice. Innovation: For the first time, we demonstrate that dRP acts intracellularly and stimulates superoxide anion generation by direct binding and activation of the NOX2 enzymatic complex. Conclusions: This study describes a novel molecular mechanism underlying the proangiogenic activity of dRP, which involves the sequential activation of NOX2 and NF-κB and upregulation of VEGFR2. Antioxid. Redox Signal. 28, 110–130.
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影响因子：
7.4
作者：
Evangelista, Alicia M.;Thompson, Melissa D.;Bolotina, Victoria M.;Tong, XiaoYong;Cohen, Richard A.
通讯作者：
Cohen, Richard A.
影响因子：
7.4
作者：
Hensley, K;Robinson, KA;Floyd, RA
通讯作者：
Floyd, RA
影响因子：
4.4
作者：
Chandel, NS;Trzyna, WC;Schumacker, PT
通讯作者：
Schumacker, PT
影响因子：
3.7
作者：
Carnesecchi, Stephanie;Carpentier, Jean-Louis;Szanto, Ildiko
通讯作者：
Szanto, Ildiko
影响因子：
120.1
作者：
Drummond, Grant R.;Selemidis, Stavros;Griendling, Kathy K.;Sobey, Christopher G.
通讯作者：
Sobey, Christopher G.