Circulation Research. 2002
Published online before print August 15, 2002, doi: 10.1161/01.RES.0000033523.08033.16
Published online before print August 15, 2002, doi: 10.1161/01.RES.0000033523.08033.16
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Submitted on April 3, 2002
Revised on July 24, 2002
Accepted on August 2, 2002
Angiotensin II Stimulation of NAD(P)H Oxidase Activity. Upstream Mediators
Puvi N. Seshiah ;
From the Division of Cardiology, Emory University, Atlanta, Ga.
* To whom correspondence should be addressed. E-mail: kgriend{at}emory.edu.
Angiotensin II (Ang II)
stimulated hypertrophy of vascular smooth muscle cells is mediated by reactive oxygen species (ROS) derived from NAD(P)H oxidases. The upstream signaling mechanisms by which Ang II activates these oxidases are unclear but may include protein kinase C, tyrosine kinases, phosphatidylinositol-3-kinase, and Rac, a small molecular weight G protein. We found that Ang IIstimulated ROS production is biphasic. The first phase occurs rapidly (peak at 30 seconds) and is dependent on protein kinase C activation. The larger second phase of ROS generation (peak at 30 minutes) requires Rac activation, because inhibition of Rac by either Clostridium difficile toxin A or dominant-negative Rac significantly inhibits Ang IIinduced ROS production. Phosphatidylinositol-3-kinase inhibitors (wortmannin or LY294002) and the epidermal growth factor (EGF) receptor kinase blocker AG1478 attenuate both Rac activation and ROS generation. The upstream activator of EGF receptor transactivation, c-Src, is also required for ROS generation, because PP1, an Src kinase inhibitor, abrogates the Ang II stimulation of both responses. These results suggest that c-Src, EGF receptor transactivation, phosphatidylinositol-3-kinase, and Rac play important roles in the sustained Ang IImediated activation of vascular smooth muscle cell NAD(P)H oxidases and provide insight into the integrated signaling mechanisms whereby Ang II stimulation leads to activation of the growth-related NAD(P)H oxidases.
Key words: angiotensin II • reactive oxygen species • vascular smooth muscle • NAD(P)H oxidase • Rac
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