Vendrov Aleksandr E, Madamanchi Nageswara R, Hakim Zeenat S, Rojas Mauricio, Runge Marschall S
Carolina Cardiovascular Biology Center, Department of Medicine, University of North Carolina, Chapel Hill, NC 27599-7055, USA.
Circ Res. 2006 May 26;98(10):1254-63. doi: 10.1161/01.RES.0000221214.37803.79. Epub 2006 Apr 6.
To characterize novel signaling pathways that underlie NAD(P)H oxidase-mediated signaling in atherosclerosis, we first examined differences in thrombin-induced gene expression between wild-type and p47phox(-/-) (NAD[P]H oxidase-deficient) VSMC. Of the 9000 genes analyzed by cDNA microarray method at the G1/S transition point, 76 genes were similarly and significantly modulated in both the cell types, whereas another 22 genes that encompass various functional groups were regulated in NAD(P)H oxidase-dependent manner. Among these 22 genes, thrombin-induced NAD(P)H oxidase-mediated regulation of Klf15, Igbp1, Ak4, Adamts5, Ech1, Serp1, Sec61a2, Aox1, Aoh1, Fxyd5, Rai14, and Serpinh1 was shown for the first time in VSMC. The role of NAD(P)H oxidase in the regulation of a subset of these genes (CD44, BMP4, Id1, and Id3) was confirmed using modulators of reactive oxygen species (ROS) generation, a ROS scavenger and in gain-of-function experiments. We then characterized regulation of these genes in restenosis and atherosclerosis. In both apoE(-/-) mice and in a mouse vascular injury model, these genes are regulated in NAD(P)H oxidase-dependent manner during vascular lesion formation. Based on these findings, we propose that NAD(P)H oxidase-dependent gene expression in general, and the CD44 and BMP4-Id signaling pathway in particular, is important in restenosis and atherosclerosis.
为了明确动脉粥样硬化中NAD(P)H氧化酶介导的信号传导所涉及的新信号通路，我们首先检测了野生型和p47phox(-/-)（NAD[P]H氧化酶缺陷型）血管平滑肌细胞（VSMC）中凝血酶诱导的基因表达差异。在G1/S转换点通过cDNA微阵列方法分析的9000个基因中，有76个基因在两种细胞类型中受到相似且显著的调节，而另外22个涵盖各种功能组的基因则以NAD(P)H氧化酶依赖性方式受到调节。在这22个基因中，凝血酶诱导的NAD(P)H氧化酶介导的对Klf15、Igbp1、Ak4、Adamts5、Ech1、Serp1、Sec61a2、Aox1、Aoh1、Fxyd5、Rai14和Serpinh1的调节首次在VSMC中得到证实。使用活性氧（ROS）生成调节剂、ROS清除剂并通过功能获得实验，证实了NAD(P)H氧化酶在这些基因（CD44、BMP4、Id1和Id3）的一个子集中的调节作用。然后，我们明确了这些基因在再狭窄和动脉粥样硬化中的调节情况。在载脂蛋白E(-/-)小鼠和小鼠血管损伤模型中，这些基因在血管病变形成过程中均以NAD(P)H氧化酶依赖性方式受到调节。基于这些发现，我们提出，一般而言，NAD(P)H氧化酶依赖性基因表达，特别是CD44和BMP4-Id信号通路，在再狭窄和动脉粥样硬化中很重要。
