Intracellular signaling pathways involved in inhibition of PAI-1 expression by CNP in endothelial cells

PAI-1 is a multifunctional protein stimulated by infectious agents and its activation is mediated by inflammatory cytokines such as TNF alpha. Recent studies demonstrate that natriuretic peptides, particularly C-type (CNP), can affect PAI-1 expression in bovine aortic smooth muscle cells and rat aortic endothelial cells. We have previously shown that CNP inhibits both basal and TNF alpha induced expression of PAI-1 in human endothelial cells. Herein, we describe mechanism by which CNP modulates signaling engaged in controlling PAI-1 expression in human endothelial cells. To examine which pathway initiated by TNF alpha is influenced, we tested kinase activity of MAP, PI3K/AKT and involvement of cGMP in endothelial cells exposed to CNP. CNP significantly increased cGMP level in endothelial cells. Its analogue, 8-Br-cGMP alone had no effect but significantly inhibited TNF alpha induced expression of PAI-1. Similarly, CNP and the inhibitors of ERK1/2 (PD098059) and PI3K (LY294002) attenuated PAI-1 expression induced by TNF alpha. CNP almost abolished TNF alpha induced phosphorylation of ERK1/2 but did not affect JNK phosphorylation, indicating that its effect on ERK1/2 was specific. These data suggest that CNP might function as the natural defense of vascular wall against cytokine induced PAI-1 release through its ability to inactivate PI3K/AKT and MEK/ERK pathways. (C) 2009 Elsevier B.V. All rights reserved.