Novel Nox inhibitor VAS2870 attenuates PDGF-dependent smooth muscle cell chemotaxis, but not proliferation
Novel Nox inhibitor VAS2870 attenuates PDGF-dependent smooth muscle cell chemotaxis, but not proliferation
复制标题
DOI：
10.1016/j.cardiores.2006.01.022
复制
发表时间：
2006-07-15
影响因子：
10.8
通讯作者：
Rosenkranz, Stephan
中科院分区：
文献类型：
--
作者：
ten Freyhaus, Henrik;Huntgeburth, Michael;Rosenkranz, Stephan
目的：NAD(P)H氧化酶(NOx)产生的活性氧(ROS)在心血管疾病的病理生理过程中起重要作用。NAD(P)H氧化酶的表达和活性受血管紧张素11和血小板衍生生长因子等生长因子的调节。我们研究了新型NOx抑制剂VAS2870对血管平滑肌细胞(VSMC)依赖PDGF的ROS释放和细胞事件的影响。方法和结果：PDGF-BB使NAD(P)H氧化酶活性(荧光素增强的化学发光)和细胞内ROS水平(用2，7-DCF共聚焦激光扫描显微镜检测)在1和2 h分别增加到229+/-9%和362+/-54%。VAS2870(10和20 mU M)预孵育可完全阻断PDGF介导的NAD(P)H氧化酶的激活和ROS的产生。由于ROS参与了生长因子诱导的多种细胞功能，因此我们研究了VAS2870对PDGF诱导的DNA合成和趋化作用的影响。PDGF以浓度依赖的方式促进VSMC迁移(改良Boyden小室，p<；0.01)和DNA合成(BrdU掺入，p<；0.01)，最大增加3.2+/-0.4倍。VAS2870(0.1-20 mU M)预孵育不影响PDGF诱导的细胞周期进程。但对PDGF依赖的VSMC趋化作用的抑制作用呈浓度依赖性(10mM时100%抑制)。这些发现与PDGF依赖的信号事件有关。蛋白印迹分析显示下游信号分子Akt、Erk和Src被PDGF激活。然而，VAS2870以浓度依赖的方式抑制PDGF依赖的Src的激活，但不抑制Akt和Erk的激活。4结论：VAS2870能有效抑制生长因子介导的VSMC释放ROS。此外，它完全抑制依赖PDGF的VSMC迁移，但不影响DNA合成。这些不同的效应反映了Src活性的关键作用，与Akt和Erk相反，Src活性似乎是氧化还原敏感的，对于PDGF诱导的趋化是绝对必要的，但不是细胞周期进展所必需的。(C)2006年欧洲心脏病学会。爱思唯尔出版，版权所有。
Objective: Reactive oxygen species (ROS) produced by NAD(P)H oxidases (Nox) play a significant role in the pathophysiology of cardiovascular diseases. Expression and activity of NAD(P)H oxidases are regulated by growth factors such as angiotensin 11 and platelet-derived growth factor (PDGF). We characterized the effects of the novel Nox inhibitor VAS2870 on PDGF-dependent ROS liberation and cellular events in vascular smooth muscle cells (VSMC).Methods and results: PDGF-BB increased NAD(P)H oxidase activity (lucigenin-enhanced chemiluminescence) and intracellular ROS levels (detected by confocal laserscanning microscopy using 2,7-DCF) to 229 +/- 9% and 362 +/- 54% at 1 and 2 h, respectively. Preincubation with VAS2870 (10 and 20 mu M) completely abolished PDGF-mediated NAD(P)H oxidase activation and ROS production. Since ROS are involved in various growth factor-induced cellular functions, the influence of VAS2870 on PDGF-induced DNA synthesis and chemotaxis was determined. PDGF promoted a 4.2 +/- 0.2-fold increase of VSMC migration (modified Boyden chamber, p < 0.01) and increased DNA synthesis by maximally 3.2 +/- 0.4-fold (BrdU incorporation, p < 0.01) in a concentration-dependent manner. Preincubation with VAS2870 (0.1-20 mu M) did not affect PDGF-induced cell cycle progression. However, it abolished PDGF-dependent chemotaxis of VSMC in a concentration-dependent manner (100% inhibition at 10 mu M). These findings were related to PDGF-dependent signaling events. Western blot analyses using phospho-specific antibodies revealed that the downstream signaling molecules Akt, Erk, and Src were activated by PDGF. However, VAS2870 blocked PDGF-dependent activation of Src, but not of Akt and Erk, in a concentration-dependent manner. \Conclusions: VAS2870 effectively suppresses growth factor-mediated ROS liberation in VSMC. Furthermore, it completely inhibits PDGF-dependent VSMC migration, whereas it does not affect DNA synthesis. These divergent effects reflect the critical role of Src activity, which-in contrast to Akt and Erk-appears to be redox-sensitive and is absolutely required for PDGF-induced chemotaxis, but not cell cycle progression. (c) 2006 European Society of Cardiology. Published by Elsevier B.V. All rights reserved.