A. Gorlach et al., A gp91phox containing NADPH oxidase selectively expressed in endothelial cells is a major source of oxygen radical generation in the arterial wall, CIRCUL RES, 87(1), 2000, pp. 26-32
Reactive oxygen species (ROS) play an important role in regulating vascular
tone and intracellular signaling; the enzymes producing ROS in the vascula
r wall are, however, poorly characterized. We investigated whether a functi
onally active NADPH oxidase similar to the leukocyte enzyme, ie, containing
the subunits p22phox and gp91phox, is expressed in endothelial cells (ECs)
and smooth muscle cells (SMCs). Phorbol 12-myristate 13-acetate (PMA), a s
timulus for leukocyte NADPH oxidase, increased ROS generation in cultured E
Cs and endothelium-intact rat aortic segments, but not in SMCs or endotheli
um-denuded arteries. NADPH enhanced chemiluminescence in all preparations.
p22phox mRNA and protein was detected in ECs and SMCs, whereas the expressi
on of gp91phox was confined to ECs. Endothelial gp91phox was identical to t
he leukocyte form as determined by sequence analysis. In contrast, mitogeni
c oxidase-1 (mox1) was expressed in SMCs, but not in ECs. To determine the
functional relevance of gp91phox expression, experiments were performed in
aortic segments from wild-type, gp91phox(-/-), and endothelial NO synthase
(eNOS)(-/-) mice. PMA-induced ROS generation was comparable in aortae from
wild-type and eNOS(-/-) mice, but was attenuated in segments from gp91phox(
-/-) mice. Endothelium-dependent relaxation was greater in aortae from gp91
phox(-/-) than from wild-type mice. The ROS scavenger tiron increased endot
helium-dependent relaxation in segments from wild-type, but not from gp91ph
ox(-/-) mice. These data demonstrate that ECs, in contrast to SMCs, express
a gp91phox-containing leukocyte-type NADPH oxidase. This enzyme is a major
source for arterial ROS generation and affects the bioavailability of endo
thelium-derived NO.