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(Stroke. 2005;36:1040.)
© 2005 American Heart Association, Inc.

Original Contributions

NAD(P)H Oxidases in Rat Basilar Arterial Endothelial Cells

Tetsuro Ago, MD; Takanari Kitazono, MD; Junya Kuroda, MD; Yasuhiro Kumai, MD; Masahiro Kamouchi, MD; Hiroaki Ooboshi, MD; Masanori Wakisaka, MD; Tsukasa Kawahara, PhD; Kazuhito Rokutan, PhD; Setsuro Ibayashi, MD Mitsuo Iida, MD

From the Department of Medicine and Clinical Science (T.A., T.Kitazono, J.K. Y.K., M.K., H.O., M.W., S.I., M.I.), Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan; and the Department of Nutrition (T.Kawahara, K.R.), School of Medicine, Tokushima University, Tokushima, Japan.

Correspondence to Tetsuro Ago, MD, PhD, Department of Medicine and Clinical Science, Graduate School of Medical Sciences, Kyushu University, Maidashi 3-1-1, Higashi-ku, Fukuoka 812-8582, Japan. E-mail agou{at}intmed2.med.kyushu-u.ac.jp

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.

Key Words: cerebrovascular disorders • endothelium • free radicals • NAD(P)H oxidase

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