Reactive oxygen species mediate the activation of Akt/protein kinase B by angiotensin II in vascular smooth muscle cells

Angiotensin II, a hypertrophic/anti-apoptotic hormone, utilizes reactive ox ygen species (ROS) as growth-related signaling molecules in Vascular smooth muscle cells (VSMCs). Recently, the cell survival protein kinase Akt/prote in kinase B (PKB) was proposed to be involved in protein synthesis. Here we show that angiotensin II causes rapid phosphorylation of Akt/PKB (6- +/- 0 .4-fold increase). Exogenous H2O2 (50-200 mu M) also stimulates Akt/PKB pho sphorylation (maximal 8- +/- 0.2-fold increase), suggesting that Akt/PKB ac tivation is redox-sensitive. Both angiotensin II and H2O2 stimulation of Ak t/PKB are abrogated by the phosphatidylinositol 3-kinase (PI3-K) inhibitors wortmannin and LY294002 (2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one) , suggesting that PI3-K is an upstream mediator of Akt/PKB activation in VS MCs, Furthermore, diphenylene iodonium, an inhibitor of flavin-containing o xidases, or overexpression of catalase to block angiotensin II-induced intr acellular H2O2 production significantly inhibits angiotensin II-induced Akt /PKB phosphorylation, indicating a role for ROS in agonist-induced Akt/PKB activation. In VSMCs infected with dominant-negative Akt/PKB, angiotensin I I-stimulated [H-3]leucine incorporation is attenuated. Thus, our studies in dicate that Akt/PKB is part of the remarkable spectrum of angiotensin II si gnaling pathways and provide insight into the highly organized signaling me chanisms coordinated by ROS, which mediate the hypertrophic response to ang iotensin II in VSMCs.