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DOI: 10.1160/TH08-05-0272
Heparan sulfate proteoglycan is essential to thrombin-induced calcium transients and nitric oxide production in aortic endothelial cells
Financial support: This study was supported in part by a grant-in-aid from the Japan Society for the promotion of Science.Publication History
Received
01 May 2008
Accepted after minor revision
30 June 2008
Publication Date:
22 November 2017 (online)
![](https://www.thieme-connect.de/media/10.1055-s-00035024/200803/lookinside/thumbnails/10-1160-th08-05-0272-1.jpg)
Summary
Thrombin induces Ca2+ transients and subsequent nitric oxide (NO) production in vascular endothelial cells. Thrombin cleaves protease-activated receptors, resulting in activation of intracellular signals, but it is not clarified how the extracellular thrombin stays around the cells to exert its enzyme activities. This study aimed to investigate the possible involvement of heparin sulfate proteoglycan (HSPG) in the effects of thrombin on vascular endothelium. Heparinase III completely removed the polysaccharide chain of HSPG in bovine aortic endothelial cells (BAECs).Thrombin induced Ca2+ transients in control BAECs, but not in heparinase III-treated BAECs. In contrast, ATP induced Ca2+ transients both in control and heparinase III-treated BAECs. Thrombin that was pre-incubated with heparin also failed to induced Ca2+ transients in BAECs. Furthermore, thrombin-induced NO production, as assessed with DAF-2 flu-orescence, was suppressed in heparinase III-treated BAECs and by the pre-incubation of thrombin with heparin. ATP-induced NO production was, however, not affected in heparinase III-treated BAECs. These results indicate that it is essential for thrombin to bind to the polysaccharide chain of HSPG for inducing Ca2+ transients and NO production in BAECs.
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References
- 1 Berkels R, Dachs C, Roesen R. et al. Simultaneous measurement of intracellular Ca2+ and nitric oxide: a new method. Cell Calcium 2000; 27: 281-286.
- 2 Martorell L, Martinez-Gonzalez J, Rodriguez C. et al. Thrombin and protease-activated receptors (PARs) in atherothrombosis. Thromb Haemost 2008; 99: 305-315.
- 3 Coughlin SR. Protease-activated receptors in hemostasis, thrombosis and vascular biology. J Thromb Haemost 2005; 03: 1800-1814.
- 4 Qiao D, Meyer K, Mundhenke C. et al. Heparan sulfate proteoglycans as regulators of fibroblast growth factor-2 signaling in brain endothelial cells. Specific role for glypican-1 in glioma angiogenesis.. J Biol Chem 2003; 278: 16045-16053.
- 5 Florian JA, Kosky JR, Ainslie K. et al. Heparan sulfate proteoglycan is a mechanosensor on endothelial cells. Circ Res 2003; 93: e136-142.
- 6 Nader HB, Dietrich CP, Buonassisi V. et al. Heparin sequences in the heparan sulfate chains of an endothelial cell proteoglycan. Proc Natl Acad Sci USA 1987; 84: 3565-3569.
- 7 Dunzendorfer S, Kaneider N, Rabensteiner A. et al. Cell-surface heparan sulfate proteoglycan-mediated regulation of human neutrophil migration by the serpin antithrombin III. Blood 2001; 97: 1079-1085.
- 8 Sheehan JP, Sadler JE. Molecular mapping of the heparin-binding exosite of thrombin. Proc Natl Acad Sci USA 1994; 91: 5518-5522.
- 9 Oike M, Ito Y. Dynamic regulation of intracellular Ca2+ concentration in aortic endothelial cells. Eur. J. Pharmacol 1997; 319: 291-298.
- 10 Kojima H, Nakatsubo N, Kikuchi K. et al. Detection and imaging of nitric oxide with novel fluorescent indicators: diaminofluoresceins. Anal Chem 1998; 70: 2446-2453.
- 11 Kimura C, Oike M, Ito Y. Acute glucose overload abolishes Ca2+ oscillation in cultured endothelial cells from bovine aorta: a possible role of superoxide anion. Circ Res 1998; 82: 677-685.
- 12 Motley ED, Eguchi K, Patterson MM. et al. Mechanism of endothelial nitric oxide synthase phosphorylation and activation by thrombin. Hypertension 2007; 49: 577-583.
- 13 Koyama T, Kimura C, Park SJ. et al. Functional implications of Ca2+ mobilizing properties for nitric oxide production in aortic endothelium. Life Sci 2002; 72: 511-520.
- 14 Tiruppathi C, Ahmmed GU, Vogel SM. et al. Ca2+ signaling, TRP channels, and endothelial permeability. Microcirculation 2006; 13: 693-708.
- 15 Viana F, de Smedt H, Droogmans G. et al. Calcium signalling through nucleotide receptor P2Y2 in cultured human vascular endothelium. Cell Calcium 1998; 24: 117-127.
- 16 Solic N, Wilson J, Wilson SJ. et al. Endothelial activation and increased heparan sulfate expression in cystic fibrosis. Am J Respir Crit Care Med 2005; 172: 892-898.
- 17 Gotte M, Bernfield M, Joussen AM. Increased leukocyte-endothelial interactions in syndecan-1-deficient mice involve heparan sulfate-dependent and -independent steps. Curr Eye Res 2005; 30: 417-422.
- 18 Hirano K. The roles of proteinase-activated receptors in the vascular physiology and pathophysiology. Arterioscl Thromb Vasc Biol 2007; 27: 27-36.
- 19 Lindahl U. Heparan sulfate-protein interactions--a concept for drug design?. Thromb Haemost 2007; 98: 109-115.