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(Circulation Research. 2001;89:408.)
© 2001 American Heart Association, Inc.

Integrative Physiology

Novel Competitive Inhibitor of NAD(P)H Oxidase Assembly Attenuates Vascular O₂^- and Systolic Blood Pressure in Mice

F. E. Rey, M. E. Cifuentes, A. Kiarash, M. T. Quinn, P. J. Pagano

From the Hypertension and Vascular Research Division (F.E.R., M.E.C., A.K., P.J.P.), Henry Ford Hospital, Detroit, Mich; and Veterinary Molecular Biology Laboratory (M.T.Q.), Montana State University, Bozeman, Mont.

Correspondence to Patrick J. Pagano, PhD, Hypertension & Vascular Research Division, Room 7044, E&R Bldg, Henry Ford Hospital, 2799 W Grand Blvd, Detroit, MI 48202-2689. E-mail ppagano1{at}hfhs.org

We previously reported enhanced expression of the p67^phox and gp91^phox components of NAD(P)H oxidase in angiotensin (Ang) II–induced hypertension, suggesting de novo assembly in response to Ang II. To examine the direct involvement of NAD(P)H oxidases in Ang II–induced O₂^- production, we designed a chimeric peptide that inhibits p47^phox association with gp91^phox in NAD(P)H oxidase (gp91ds-tat). This was achieved by linking a 9-amino acid peptide (aa) derived from HIV-coat protein (tat) to a 9-aa sequence of gp91^phox (known to interact with p47^phox). As a control, we constructed a chimera containing tat and a scrambled gp91 sequence (scramb-tat). We found that gp91ds-tat decreased O₂^- levels in aortic rings treated with Ang II (10 pmol/L) but had no effect on either the O₂^--generating enzyme xanthine oxidase or potassium superoxide–generated O₂^-. We infused vehicle, Ang II (0.75 mg · kg^-1 · d^-1), Ang II+gp91ds-tat (10 mg · kg^-1 · d^-1), or Ang II+scramb-tat intraperitoneally in C57Bl/6 mice and measured systolic blood pressure (SBP) on days 0, 3, 5, and 7 of infusion. SBP increased by day 3 in mice given Ang II and Ang II+scramb-tat but was significantly lower with Ang II+gp91-tat. On day 7, SBP was still significantly inhibited in mice given Ang II+gp91ds-tat, whereas Ang II–induced O₂^- production was inhibited throughout the aorta as detected by dihydroethidium staining, consistent with the ability of this inhibitor to block the various vascular NAD(P)H oxidase isoforms. These data support the hypothesis that inhibition of the interaction of p47^phox and gp91^phox (or its homologues) can block O₂^- production and attenuate blood pressure elevation in mice.

Key Words: superoxide • angiotensin II • NAD(P)H oxidase • gp91^phox • p47^phox

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				B. Lassegue and R. E. Clempus Vascular NAD(P)H oxidases: specific features, expression, and regulation Am J Physiol Regulatory Integrative Comp Physiol, August 1, 2003; 285(2): R277 - R297. [Abstract] [Full Text] [PDF]

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				J. Liu, F. Yang, X.-P. Yang, M. Jankowski, and P. J. Pagano NAD(P)H Oxidase Mediates Angiotensin II-Induced Vascular Macrophage Infiltration and Medial Hypertrophy Arterioscler Thromb Vasc Biol, May 1, 2003; 23(5): 776 - 782. [Abstract] [Full Text] [PDF]

				R. P. Brandes A Radical Adventure: The Quest for Specific Functions and Inhibitors of Vascular NAPDH Oxidases Circ. Res., April 4, 2003; 92(6): 583 - 585. [Full Text] [PDF]

				G. M. Jacobson, H. M. Dourron, J. Liu, O. A. Carretero, D. J. Reddy, T. Andrzejewski, and P. J. Pagano Novel NAD(P)H Oxidase Inhibitor Suppresses Angioplasty-Induced Superoxide and Neointimal Hyperplasia of Rat Carotid Artery Circ. Res., April 4, 2003; 92(6): 637 - 643. [Abstract] [Full Text] [PDF]

				J.-M. Li and A. M. Shah Mechanism of Endothelial Cell NADPH Oxidase Activation by Angiotensin II. ROLE OF THE p47phox SUBUNIT J. Biol. Chem., March 28, 2003; 278(14): 12094 - 12100. [Abstract] [Full Text] [PDF]

				P. T. Schumacker Angiotensin II Signaling in the Brain: Compartmentalization of Redox Signaling? Circ. Res., November 29, 2002; 91(11): 982 - 984. [Full Text] [PDF]

				F. E. Rey, X.-C. Li, O. A. Carretero, J. L. Garvin, and P. J. Pagano Perivascular Superoxide Anion Contributes to Impairment of Endothelium-Dependent Relaxation: Role of gp91phox Circulation, November 5, 2002; 106(19): 2497 - 2502. [Abstract] [Full Text] [PDF]

				U. Landmesser, H. Cai, S. Dikalov, L. McCann, J. Hwang, H. Jo, S. M. Holland, and D. G. Harrison Role of p47phox in Vascular Oxidative Stress and Hypertension Caused by Angiotensin II Hypertension, October 1, 2002; 40(4): 511 - 515. [Abstract] [Full Text] [PDF]

				F. Krotz, H. Y. Sohn, T. Gloe, S. Zahler, T. Riexinger, T. M. Schiele, B. F. Becker, K. Theisen, V. Klauss, and U. Pohl NAD(P)H oxidase-dependent platelet superoxide anion release increases platelet recruitment Blood, July 18, 2002; 100(3): 917 - 924. [Abstract] [Full Text] [PDF]

				A. Csiszar, Z. Ungvari, J. G. Edwards, P. Kaminski, M. S. Wolin, A. Koller, and G. Kaley Aging-Induced Phenotypic Changes and Oxidative Stress Impair Coronary Arteriolar Function Circ. Res., June 14, 2002; 90(11): 1159 - 1166. [Abstract] [Full Text] [PDF]

				R. M. Touyz, X. Chen, F. Tabet, G. Yao, G. He, M. T. Quinn, P. J. Pagano, and E. L. Schiffrin Expression of a Functionally Active gp91phox-Containing Neutrophil-Type NAD(P)H Oxidase in Smooth Muscle Cells From Human Resistance Arteries: Regulation by Angiotensin II Circ. Res., June 14, 2002; 90(11): 1205 - 1213. [Abstract] [Full Text] [PDF]

				B. Lassegue and K. K. Griendling Out Phoxing the Endothelium: What's Left Without p47? Circ. Res., February 8, 2002; 90(2): 123 - 124. [Full Text] [PDF]