NAD(P)H oxidases in rat basilar arterial endothelial cells
NAD(P)H oxidases in rat basilar arterial endothelial cells
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DOI：
10.1161/01.str.0000163111.05825.0b
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发表时间：
2005-05-01
期刊：
影响因子：
8.3
通讯作者：
Iida, M
中科院分区：
文献类型：
--
作者：
Ago, T;Kitazono, T;Iida, M
背景和目的-活性氧（ROS）可能在血管张力的调节和血管疾病（如中风）的发展中起关键作用。NAD（P）H氧化酶是血管细胞（包括内皮细胞）中ROS的主要来源。据认为，Nox 2和Nox 4在Nox同系物中仅在非脑血管的内皮细胞中表达。然而，脑动脉内皮细胞中NAD（P）H氧化酶的精确分子身份尚未完全了解。我们研究了Nox同源物的表达及其在脑动脉内皮细胞中的激活机制。方法-我们分离和培养的基底动脉内皮细胞（BAECs）的Sprague-Dawley大鼠。结果RT-PCR显示BAEC中Nox 4高表达，边缘表达Nox 2。此外，Nox 1在BAEC中在mRNA和蛋白水平上均高度表达。免疫组织化学染色显示基底动脉内皮细胞中Nox 1的表达显著。对于NAD（P）H氧化酶的胞浆成分，BAEC表达p67（phox），并在较小程度上表达p47（phox）、Noxo 1和Noxa 1。NADH和NADPH均能诱导BAEC膜产生超氧化物。吞噬细胞型胞质组分p47（phox）和p67（phox）显著增强了BAEC膜的NADPH诱导的超氧化物产生，而这些组分未能增加NADPH诱导的超氧化物产生。脑动脉内皮细胞NAD（P）H氧化酶可能具有独特的被吞噬细胞型胞质组分激活的机制。
Background and Purpose - Reactive oxygen species (ROS) may play a critical role in the regulation of vascular tone and development of vascular diseases, such as stroke. NAD( P) H oxidase is a major source of ROS in vascular cells, including endothelial cells. It has been considered that Nox2 and Nox4 are exclusively expressed among Nox homologues in the endothelial cells of noncerebral blood vessels. However, the precise molecular identity of the NAD( P) H oxidase in the endothelial cells of the cerebral arteries is not fully understood. We examined the expression of Nox homologues and their activation mechanism in the endothelial cells of the cerebral arteries.Methods - We isolated and cultured basilar artery endothelial cells (BAECs) of Sprague-Dawley rats. Expression of NAD( P) H oxidase was examined by reverse-transcription-polymerase chain reaction (RT-PCR) and immunohistological staining.Results - RT-PCR disclosed abundant expression of Nox4 with marginal Nox2 in BAEC. In addition, Nox1 was expressed highly both at mRNA and protein levels in BAECs. Immunohistological staining also showed the prominent expression of Nox1 in the endothelial cells of the basilar artery. With respect to the cytosolic components of NAD( P) H oxidases, BAECs expressed p67(phox) and, to a lesser extent, p47(phox), Noxo1, and Noxa1. Both NADH and NADPH induced superoxide production of the BAEC membranes. The phagocyte-type cytosolic components, p47(phox) and p67(phox), significantly enhanced the NADH-induced superoxide production of the BAEC membranes, whereas the components failed to increase the NADPH-induced superoxide production.Conclusions - Nox1 is highly expressed in the endothelial cells of the cerebral arteries along with Nox2 and Nox4, and the endothelial NAD( P) H oxidase of the cerebral arteries may have a unique activation mechanism by the phagocyte-type cytosolic components.