TY - JOUR
T1 - Heparan sulfate proteoglycan is essential to thrombin-induced calcium transients and nitric oxide production in aortic endothelial cells
AU - Kimura, Chiwaka
AU - Oike, Masahiro
N1 - Copyright:
Copyright 2008 Elsevier B.V., All rights reserved.
PY - 2008/9
Y1 - 2008/9
N2 - 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 fluorescence, 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 Ca 2+ transients and NO production in BAECs.
AB - 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 fluorescence, 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 Ca 2+ transients and NO production in BAECs.
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U2 - 10.1160/TH08-05-0272
DO - 10.1160/TH08-05-0272
M3 - Article
C2 - 18766266
AN - SCOPUS:51349109184
VL - 100
SP - 483
EP - 488
JO - Thrombosis and Haemostasis
JF - Thrombosis and Haemostasis
SN - 0340-6245
IS - 3
ER -