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* Institut für Kardiovaskuläre Physiologie, Klinikum der J. W. Goethe-Universität, 60590 Frankfurt/Main, Germany; and
§ Stiftung Deutsche Klinik für Diagnostik, Fachbereich Hämostaseologie, 65191 Wiesbaden, Germany
1Correspondence: Institut für Kardiovaskuläre Physiologie. Klinikum der J. W. Goethe-Universität, Theodor-Stern-Kai 7, D-60590 Frankfurt/Main, Germany. E-mail: A.Goerlach{at}em.uni-frankfurt.de
Vascular injury after balloon angioplasty results in the rapid activation of platelets leading to the release of growth factors and vasoactive substances. In addition, up-regulation of tissue factor (TF) and an increased production of reactive oxygen species (ROS) have been detected at sites of vascular injury. We investigated whether platelet-derived products (PDP) released from activated human platelets increase ROS production, resulting in the induction of TF expression in vascular smooth muscle cells (SMC). PDP induced a time- and concentration-dependent increase in ROS generation in cultured SMC that was mediated mainly by PDGF-AB and TGF-ß1 and impaired by the flavin inhibitor diphenylene iodonium. Increased ROS formation was associated with enhanced mRNA levels of the small NAD(P)H oxidase subunit p22phox or its smooth muscle isoform. Transient transfection with a p22phox antisense vector decreased PDP-induced ROS generation. PDP up-regulated TF mRNA expression, which was redox sensitive and reduced by transfection of the p22phox antisense vector. In addition, PDP-stimulated reporter gene activity of two TF promoter constructs was decreased by coexpression of the p22phox antisense vector. These results indicate that activated platelets up-regulate TF expression and that this response involves ROS generation and a p22phox-containing NAD(P)H oxidase in SMC.—Görlach, A., Brandes, R. P., Bassus, S., Kronemann, N., Kirchmaier, C. M., Busse, R., Schini-Kerth, V. B. Oxidative stress and expression of p22phox are involved in the up-regulation of tissue factor in vascular smooth muscle cells in response to activated platelets.
Key Words: NAD(P)H oxidase • reactive oxygen species • platelet-derived products • vascular injury • antisense technique
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