Abstract

In contrast to other cell types, vascular smooth muscle cells modify their phenotype in response to external signals. NADPH oxidase 4 (Nox4) is critical for maintenance of smooth muscle gene expression; however, the underlying mechanisms are incompletely characterized. Using smooth muscle alpha-actin (SMA) as a prototypical smooth muscle gene and transforming growth factor-beta (TGF-beta) as a differentiating agent, we examined Nox4-dependent signaling. TGF-beta increases Nox4 expression and activity in human aortic smooth muscle cells (HASMC). Transfection of HASMC with siRNA against Nox4 (siNox4) abolishes TGF-beta-induced SMA expression and stress fiber formation. siNox4 also significantly inhibits TGF-beta-stimulated p38MAPK phosphorylation, as well as that of its substrate, mitogen-activated protein kinase-activated protein kinase-2. Moreover, the p38MAPK inhibitor SB-203580 nearly completely blocks the SMA increase induced by TGF-beta. Inhibition of either p38MAPK or NADPH oxidase-derived reactive oxygen species impairs the TGF-beta-induced phosphorylation of Ser103 on serum response factor (SRF) and reduces its transcriptional activity. Binding of SRF to myocardin-related transcription factor (MRTF) is also necessary, because downregulation of MRTF by siRNA abolishes TGF-beta-induced SMA expression. Taken together, these data suggest that Nox4 regulates SMA expression via activation of a p38MAPK/SRF/MRTF pathway in response to TGF-beta. (C) 2010 Elsevier Inc. All rights reserved.