From the Vascular Biology (P.J.P., J.K.C.) and Myocardial Biology
(D.A.S., W.S.C.) Laboratories, Boston Medical Center, Boston, Mass; and the
Pediatric Oncology Branch, National Cancer Institute, National Institutes of
Health, Bethesda, Md (S.J.C.).
Abstract—Superoxide radical
(O2-) is ubiquitously critical to the
bioactivity of endothelial nitric oxide. In
angiotensin-dependent hypertension, vascular
O2- levels rise and impede
endothelium/nitric oxide–dependent vascular
relaxation. We have reported that the major
O2- source in the rabbit aorta is adventitial
fibroblast phagocyte-like NADPH oxidase and shown that
angiotensin (Ang) II treatment of adventitial fibroblasts
causes a concentration-dependent increase in particulate
NADPH-dependent O2-. From cultured rabbit
aortic adventitial fibroblasts treated or not treated with Ang II, we
prepared particulate fractions and measured lucigenin-enhanced
chemiluminescence. Because [Sar1,Thr8]-Ang
II, a generalized antagonist of Ang II and plausible
inhibitor of the conversion of Ang II, reversed Ang II (10
nmol/L)–induced NADH- and NADPH-dependent O2-
to basal levels, we tested the effect of the inhibitor of
aminopeptidase N, amastatin (10 µmol/L), and
found no effect on Ang II–stimulated O2-.
Ang(1-7), Ang III, and Ang IV also were not effective in stimulating
O2- levels at concentrations similar to those
of Ang II. Kinetic analysis showed a rise in NADPH oxidase
O2- production in response to Ang II,
which peaks at 3 hours and returns to basal levels by 16 hours.
p67phox, a cytosolic factor, appears to be affected at both
the level of transcription and protein synthesis because actinomycin
and cycloheximide individually inhibited the observed effect. A partial
sequence of p67phox was recovered by reverse transcriptase
from mRNA harvested from cultured rabbit aortic adventitial
fibroblasts. Furthermore, the p67phox mRNA transcript in
aortic fibroblasts is induced by Ang II before the peak of NADPH
oxidase by Northern analysis and ribonuclease protection
assays. These data suggest that Ang II stimulates NAD(P)H oxidase
O2- generation in fibroblasts of aortic
adventitia via transcriptional activation of p67phox. These
data also provide preliminary evidence for the regulation of factors of
the NADPH oxidase and potentially provide a novel means by which to
abrogate the development of O2--dependent
hypertension.
© 1998 American Heart Association, Inc.
Scientific Contributions
Angiotensin II Induces p67phox mRNA Expression and NADPH Oxidase Superoxide Generation in Rabbit Aortic Adventitial Fibroblasts
Key Words: rabbits • angiotensin II • superoxide • free radicals • reactive oxygen species • NADPH oxidoreductases
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B. Lassegue, D. Sorescu, K. Szocs, Q. Yin, M. Akers, Y. Zhang, S. L. Grant, J. D. Lambeth, and K. K. Griendling Novel gp91phox Homologues in Vascular Smooth Muscle Cells : nox1 Mediates Angiotensin II-Induced Superoxide Formation and Redox-Sensitive Signaling Pathways Circ. Res., May 11, 2001; 88(9): 888 - 894. [Abstract] [Full Text] [PDF]
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F. E. Rey, M. E. Cifuentes, A. Kiarash, M. T. Quinn, and P. J. Pagano Novel Competitive Inhibitor of NAD(P)H Oxidase Assembly Attenuates Vascular O2- and Systolic Blood Pressure in Mice Circ. Res., August 31, 2001; 89(5): 408 - 414. [Abstract] [Full Text] [PDF]
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