Circulation Research, Vol 74, 1141-1148, Copyright © 1994 by American Heart Association

ARTICLES

Angiotensin II stimulates NADH and NADPH oxidase activity in cultured vascular smooth muscle cells

KK Griendling, CA Minieri, JD Ollerenshaw and RW Alexander
Division of Cardiology, Emory University, Atlanta, GA 30322.

The signaling pathways involved in the long-term metabolic effects of angiotensin II (Ang II) in vascular smooth muscle cells are incompletely understood but include the generation of molecules likely to affect oxidase activity. We examined the ability of Ang II to stimulate superoxide anion formation and investigated the identity of the oxidases responsible for its production. Treatment of vascular smooth muscle cells with Ang II for 4 to 6 hours caused a 2.7 +/- 0.4- fold increase in intracellular superoxide anion formation as detected by lucigenin assay. This superoxide appeared to result from activation of both the NADPH and NADH oxidases. NADPH oxidase activity increased from 3.23 +/- 0.61 to 11.80 +/- 1.72 nmol O2-/min per milligram protein after 4 hours of Ang II, whereas NADH oxidase activity increased from 16.76 +/- 2.13 to 45.00 +/- 4.57 nmol O2-/min per milligram protein. The NADPH oxidase activity was stimulated by exogenous phosphatidic and arachidonic acids and was partially inhibited by the specific inhibitor diphenylene iodinium. NADH oxidase activity was increased by arachidonic and linoleic acids, was insensitive to exogenous phosphatidic acid, and was inhibited by high concentrations of quinacrine. Both of these oxidases appear to reside in the plasma membrane, on the basis of migration of the activity after cellular fractionation and their apparent insensitivity to the mitochondrial poison KCN. These observations suggest that Ang II specifically activates enzyme systems that promote superoxide generation and raise the possibility that these pathways function as second messengers for long-term responses, such as hypertrophy or hyperplasia.

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				P. Chen, A. M. Guo, P. A. Edwards, G. Trick, and A. G. Scicli Role of NADPH oxidase and ANG II in diabetes-induced retinal leukostasis Am J Physiol Regulatory Integrative Comp Physiol, October 1, 2007; 293(4): R1619 - R1629. [Abstract] [Full Text] [PDF]

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				F. J. Miller Jr, M. Filali, G. J. Huss, B. Stanic, A. Chamseddine, T. J. Barna, and F. S. Lamb Cytokine Activation of Nuclear Factor {kappa}B in Vascular Smooth Muscle Cells Requires Signaling Endosomes Containing Nox1 and ClC-3 Circ. Res., September 28, 2007; 101(7): 663 - 671. [Abstract] [Full Text] [PDF]

				F. Anan, T. Masaki, Y. Umeno, T. Iwao, H. Yonemochi, N. Eshima, T. Saikawa, and H. Yoshimatsu Correlations of high-sensitivity C-reactive protein and atherosclerosis in Japanese type 2 diabetic patients Eur. J. Endocrinol., September 1, 2007; 157(3): 311 - 317. [Abstract] [Full Text] [PDF]

				S. Choudhary, M. Lu, R. Cui, and A. R. Brasier Involvement of a Novel Rac/RhoA Guanosine Triphosphatase-Nuclear Factor-{kappa}B Inducing Kinase Signaling Pathway Mediating Angiotensin II-Induced RelA Transactivation Mol. Endocrinol., September 1, 2007; 21(9): 2203 - 2217. [Abstract] [Full Text] [PDF]

				A. Sachse and G. Wolf Angiotensin II Induced Reactive Oxygen Species and the Kidney J. Am. Soc. Nephrol., September 1, 2007; 18(9): 2439 - 2446. [Abstract] [Full Text] [PDF]

				A. Katsurada, Y. Hagiwara, K. Miyashita, R. Satou, K. Miyata, N. Ohashi, L. G. Navar, and H. Kobori Novel sandwich ELISA for human angiotensinogen Am J Physiol Renal Physiol, September 1, 2007; 293(3): F956 - F960. [Abstract] [Full Text] [PDF]

				H. Kobori, Y. Ozawa, R. Satou, A. Katsurada, K. Miyata, N. Ohashi, N. Hase, Y. Suzaki, C. D. Sigmund, and L. G. Navar Kidney-specific enhancement of ANG II stimulates endogenous intrarenal angiotensinogen in gene-targeted mice Am J Physiol Renal Physiol, September 1, 2007; 293(3): F938 - F945. [Abstract] [Full Text] [PDF]

				M. J. Haurani and P. J. Pagano Adventitial fibroblast reactive oxygen species as autacrine and paracrine mediators of remodeling: Bellwether for vascular disease? Cardiovasc Res, September 1, 2007; 75(4): 679 - 689. [Abstract] [Full Text] [PDF]

				S. J. An, R. Boyd, M. Zhu, A. Chapman, D. R. Pimentel, and H. D. Wang NADPH oxidase mediates angiotensin II-induced endothelin-1 expression in vascular adventitial fibroblasts Cardiovasc Res, September 1, 2007; 75(4): 702 - 709. [Abstract] [Full Text] [PDF]

				W Zhao, D I Diz, and M E Robbins Oxidative damage pathways in relation to normal tissue injury Br. J. Radiol., September 1, 2007; 80(Special_Issue_1): S23 - S31. [Abstract] [Full Text] [PDF]

				Y. Ozawa and H. Kobori Crucial role of Rho-nuclear factor-{kappa}B axis in angiotensin II-induced renal injury Am J Physiol Renal Physiol, July 1, 2007; 293(1): F100 - F109. [Abstract] [Full Text] [PDF]

				A. A. Miller, G. R. Drummond, A. E. Mast, H. H.H.W. Schmidt, and C. G. Sobey Effect of Gender on NADPH-Oxidase Activity, Expression, and Function in the Cerebral Circulation: Role of Estrogen Stroke, July 1, 2007; 38(7): 2142 - 2149. [Abstract] [Full Text] [PDF]