Novel blocker of NFAT activation inhibits IL-6 production in human myometrial arteries and reduces vascular smooth muscle cell proliferation
Open Access
- 1 March 2007
- journal article
- Published by American Physiological Society in American Journal of Physiology-Cell Physiology
- Vol. 292 (3), C1167-C1178
- https://doi.org/10.1152/ajpcell.00590.2005
Abstract
The calcineurin/nuclear factor of activated T cells (NFAT) signaling pathway has been found to play a role in regulating growth and differentiation in several cell types. However, the functional significance of NFAT in the vasculature is largely unclear. Here we show that NFATc1, NFATc3, and NFATc4 are expressed in human myometrial arteries. Confocal immunofluorescence and Western blot analysis revealed that endothelin-1 efficiently increases NFATc3 nuclear accumulation in native arteries. Endothelin-1 also stimulates NFAT-dependent transcriptional activity, as shown by a luciferase reporter assay. Both the agonist-induced NFAT nuclear accumulation and transcriptional activity were prevented by the calcineurin inhibitor CsA and by the novel NFAT blocker A-285222. Chronic inhibition of NFAT significantly reduced IL-6 production in intact myometrial arteries and inhibited cell proliferation in vascular smooth muscle cells cultured from explants from the same arteries. Furthermore, by using small interfering RNA-mediated reduction of NFATc3, we show that this isoform is involved in the regulation of cell proliferation. Protein synthesis in intact arteries was investigated using autoradiography of [35S]methionine incorporation in serum-free culture. Inhibition of NFAT signaling did not affect overall protein synthesis or specifically the synthesis rates of major proteins associated with the contractile/cytoskeletal system. An intact contractile phenotype under these conditions was also shown by unchanged force response to depolarization or agonist stimulation. Our results demonstrate NFAT expression and activation in native human vessels and point out A-285222 as a powerful pharmacological blocker of NFAT signaling in the vasculature.Keywords
This publication has 39 references indexed in Scilit:
- High Glucose Activates Nuclear Factor of Activated T Cells in Native Vascular Smooth MuscleArteriosclerosis, Thrombosis, and Vascular Biology, 2006
- Nuclear Factor of Activated T Cells and Serum Response Factor Cooperatively Regulate the Activity of an α-Actin Intronic EnhancerOnline Journal of Public Health Informatics, 2005
- Blockade of Nuclear Factor of Activated T Cells Activation Signaling Suppresses Balloon Injury-induced Neointima Formation in a Rat Carotid Artery ModelOnline Journal of Public Health Informatics, 2005
- A potential role for nuclear factor of activated T-cells in receptor tyrosine kinase and G-protein-coupled receptor agonist-induced cell proliferationBiochemical Journal, 2002
- Opposing Actions of Inositol 1,4,5-Trisphosphate and Ryanodine Receptors on Nuclear Factor of Activated T-cells Regulation in Smooth MuscleOnline Journal of Public Health Informatics, 2002
- Calcineurin-GATA-6 pathway is involved in smooth muscle–specific transcriptionThe Journal of cell biology, 2002
- Increased Store-Operated Ca2+ Entry into Contractile Vascular Smooth Muscle following Organ CultureJournal of Vascular Research, 2001
- Generic Signals and Specific Outcomes: Signaling through Ca2+, Calcineurin, and NF-ATCell, 1999
- TRANSCRIPTION FACTORS OF THE NFAT FAMILY:Regulation and FunctionAnnual Review of Immunology, 1997
- The Mechanism of Action of Cyclosporin A and FK506Clinical Immunology and Immunopathology, 1996