Abstract

1 The aim of this study was to identify the presence of matrix metalloproteinase-9 (MMP-9) in human platelets and systematically examine its inhibitory mechanisms of platelet activation. 2 In this study, we report on an efficient method for the quantitative analysis of pro-MMP-9 in human platelets using capillary zone electrophoresis (CZE). To elucidate subcellular localization of MMP-9 in human platelets, we investigated intraplatelet MMP-9 by immunogold labeling and visualized it using electron microscopy. In an in vivo thrombotic study, platelet thrombus formation was induced by irradiation of mesenteric venules with filtered light in mice pretreated with fluorescein sodium. 3 MMP-9-gold labeling was observed on the plasma membrane, α-granules, open canalicular system, and within the cytoplasma both in resting and activated platelets. Furthermore, activated MMP-9 concentration-dependently (15-90 ng ml -1) inhibited platelet aggregation stimulated by agonists. Activated MMP-9 (21 and 90 ng ml -1) inhibited phosphoinositide breakdown, intracellular Ca 2+ mobilization, and thromboxane A 2 formation in human platelets stimulated by collagen (1 μg ml -1). In addition, activated MMP-9 (21 and 90 ng ml -1) significantly increased the formation of nitric oxide/cyclic GMP. 4 Rapid phosphorylation of a platelet protein of Mr 47,000 (P47), a marker of protein kinase C activation, was triggered by phorbol-12, 13-dibutyrate (PDBu) (60 nM). This phosphorylation was markedly inhibited by activated MMP-9 (21 and 90 ng ml -1). Activated MMP-9 (1 μg g -1) significantly prolonged the latency period of inducing platelet plug formation in mesenteric venules. 5 These results indicate that the antiplatelet activity of activated MMP-9 may be involved in the following pathways. (1) Activated MMP-9 may inhibit the activation of phospholipase C, followed by inhibition of phosphoinositide breakdown, protein kinase C activation, and thromboxane A 2 formation, thereby leading to inhibition of intracellular Ca 2+ mobilization. (2) Activated MMP-9 also activated the formation of nitric oxide/cyclic GMP, resulting in inhibition of platelet aggregation. These results strongly indicate that MMP-9 is a potent inhibitor of aggregation. It may play an important role as a negative feedback regulator during platelet activation.

Original languageEnglish
Pages (from-to)193-201
Number of pages9
JournalBritish Journal of Pharmacology
Volume143
Issue number1
DOIs
Publication statusPublished - Sep 2004

Fingerprint

Matrix Metalloproteinase 9
Platelet Activation
Blood Platelets
Venules
Thromboxanes
Cyclic GMP
Phosphatidylinositols
Platelet Aggregation
varespladib methyl
Protein Kinase C
Nitric Oxide
human MMP9 protein
In Vitro Techniques
Phosphorylation
Phorbol 12,13-Dibutyrate
Matrix Metalloproteinase 1
Capillary Electrophoresis
Type C Phospholipases
Fluorescein
Gold

Keywords

  • Arterial thrombosis
  • Immunogold
  • Matrix metalloproteinase-9
  • Platelet aggregation
  • Protein kinase C

ASJC Scopus subject areas

  • Pharmacology

Cite this

Expression of matrix metalloproteinase-9 in human platelets : Regulation of platelet activation in in vitro and in vivo studies. / Sheu, Joen-Rong; Fong, Tsorng-Harn; Liu, Cheng-Ming; Shen, Ming Y.; Chen, Ta-Liang; Chang, Yi; Lu, Meng S.; Hsiao, George.

In: British Journal of Pharmacology, Vol. 143, No. 1, 09.2004, p. 193-201.

Research output: Contribution to journalArticle

@article{2174026dc00f4ca6a60b469b03e6673d,
title = "Expression of matrix metalloproteinase-9 in human platelets: Regulation of platelet activation in in vitro and in vivo studies",
abstract = "1 The aim of this study was to identify the presence of matrix metalloproteinase-9 (MMP-9) in human platelets and systematically examine its inhibitory mechanisms of platelet activation. 2 In this study, we report on an efficient method for the quantitative analysis of pro-MMP-9 in human platelets using capillary zone electrophoresis (CZE). To elucidate subcellular localization of MMP-9 in human platelets, we investigated intraplatelet MMP-9 by immunogold labeling and visualized it using electron microscopy. In an in vivo thrombotic study, platelet thrombus formation was induced by irradiation of mesenteric venules with filtered light in mice pretreated with fluorescein sodium. 3 MMP-9-gold labeling was observed on the plasma membrane, α-granules, open canalicular system, and within the cytoplasma both in resting and activated platelets. Furthermore, activated MMP-9 concentration-dependently (15-90 ng ml -1) inhibited platelet aggregation stimulated by agonists. Activated MMP-9 (21 and 90 ng ml -1) inhibited phosphoinositide breakdown, intracellular Ca 2+ mobilization, and thromboxane A 2 formation in human platelets stimulated by collagen (1 μg ml -1). In addition, activated MMP-9 (21 and 90 ng ml -1) significantly increased the formation of nitric oxide/cyclic GMP. 4 Rapid phosphorylation of a platelet protein of Mr 47,000 (P47), a marker of protein kinase C activation, was triggered by phorbol-12, 13-dibutyrate (PDBu) (60 nM). This phosphorylation was markedly inhibited by activated MMP-9 (21 and 90 ng ml -1). Activated MMP-9 (1 μg g -1) significantly prolonged the latency period of inducing platelet plug formation in mesenteric venules. 5 These results indicate that the antiplatelet activity of activated MMP-9 may be involved in the following pathways. (1) Activated MMP-9 may inhibit the activation of phospholipase C, followed by inhibition of phosphoinositide breakdown, protein kinase C activation, and thromboxane A 2 formation, thereby leading to inhibition of intracellular Ca 2+ mobilization. (2) Activated MMP-9 also activated the formation of nitric oxide/cyclic GMP, resulting in inhibition of platelet aggregation. These results strongly indicate that MMP-9 is a potent inhibitor of aggregation. It may play an important role as a negative feedback regulator during platelet activation.",
keywords = "Arterial thrombosis, Immunogold, Matrix metalloproteinase-9, Platelet aggregation, Protein kinase C",
author = "Joen-Rong Sheu and Tsorng-Harn Fong and Cheng-Ming Liu and Shen, {Ming Y.} and Ta-Liang Chen and Yi Chang and Lu, {Meng S.} and George Hsiao",
year = "2004",
month = "9",
doi = "10.1038/sj.bjp.0705917",
language = "English",
volume = "143",
pages = "193--201",
journal = "British Journal of Pharmacology",
issn = "0007-1188",
publisher = "John Wiley and Sons Inc.",
number = "1",

}

TY - JOUR

T1 - Expression of matrix metalloproteinase-9 in human platelets

T2 - Regulation of platelet activation in in vitro and in vivo studies

AU - Sheu, Joen-Rong

AU - Fong, Tsorng-Harn

AU - Liu, Cheng-Ming

AU - Shen, Ming Y.

AU - Chen, Ta-Liang

AU - Chang, Yi

AU - Lu, Meng S.

AU - Hsiao, George

PY - 2004/9

Y1 - 2004/9

N2 - 1 The aim of this study was to identify the presence of matrix metalloproteinase-9 (MMP-9) in human platelets and systematically examine its inhibitory mechanisms of platelet activation. 2 In this study, we report on an efficient method for the quantitative analysis of pro-MMP-9 in human platelets using capillary zone electrophoresis (CZE). To elucidate subcellular localization of MMP-9 in human platelets, we investigated intraplatelet MMP-9 by immunogold labeling and visualized it using electron microscopy. In an in vivo thrombotic study, platelet thrombus formation was induced by irradiation of mesenteric venules with filtered light in mice pretreated with fluorescein sodium. 3 MMP-9-gold labeling was observed on the plasma membrane, α-granules, open canalicular system, and within the cytoplasma both in resting and activated platelets. Furthermore, activated MMP-9 concentration-dependently (15-90 ng ml -1) inhibited platelet aggregation stimulated by agonists. Activated MMP-9 (21 and 90 ng ml -1) inhibited phosphoinositide breakdown, intracellular Ca 2+ mobilization, and thromboxane A 2 formation in human platelets stimulated by collagen (1 μg ml -1). In addition, activated MMP-9 (21 and 90 ng ml -1) significantly increased the formation of nitric oxide/cyclic GMP. 4 Rapid phosphorylation of a platelet protein of Mr 47,000 (P47), a marker of protein kinase C activation, was triggered by phorbol-12, 13-dibutyrate (PDBu) (60 nM). This phosphorylation was markedly inhibited by activated MMP-9 (21 and 90 ng ml -1). Activated MMP-9 (1 μg g -1) significantly prolonged the latency period of inducing platelet plug formation in mesenteric venules. 5 These results indicate that the antiplatelet activity of activated MMP-9 may be involved in the following pathways. (1) Activated MMP-9 may inhibit the activation of phospholipase C, followed by inhibition of phosphoinositide breakdown, protein kinase C activation, and thromboxane A 2 formation, thereby leading to inhibition of intracellular Ca 2+ mobilization. (2) Activated MMP-9 also activated the formation of nitric oxide/cyclic GMP, resulting in inhibition of platelet aggregation. These results strongly indicate that MMP-9 is a potent inhibitor of aggregation. It may play an important role as a negative feedback regulator during platelet activation.

AB - 1 The aim of this study was to identify the presence of matrix metalloproteinase-9 (MMP-9) in human platelets and systematically examine its inhibitory mechanisms of platelet activation. 2 In this study, we report on an efficient method for the quantitative analysis of pro-MMP-9 in human platelets using capillary zone electrophoresis (CZE). To elucidate subcellular localization of MMP-9 in human platelets, we investigated intraplatelet MMP-9 by immunogold labeling and visualized it using electron microscopy. In an in vivo thrombotic study, platelet thrombus formation was induced by irradiation of mesenteric venules with filtered light in mice pretreated with fluorescein sodium. 3 MMP-9-gold labeling was observed on the plasma membrane, α-granules, open canalicular system, and within the cytoplasma both in resting and activated platelets. Furthermore, activated MMP-9 concentration-dependently (15-90 ng ml -1) inhibited platelet aggregation stimulated by agonists. Activated MMP-9 (21 and 90 ng ml -1) inhibited phosphoinositide breakdown, intracellular Ca 2+ mobilization, and thromboxane A 2 formation in human platelets stimulated by collagen (1 μg ml -1). In addition, activated MMP-9 (21 and 90 ng ml -1) significantly increased the formation of nitric oxide/cyclic GMP. 4 Rapid phosphorylation of a platelet protein of Mr 47,000 (P47), a marker of protein kinase C activation, was triggered by phorbol-12, 13-dibutyrate (PDBu) (60 nM). This phosphorylation was markedly inhibited by activated MMP-9 (21 and 90 ng ml -1). Activated MMP-9 (1 μg g -1) significantly prolonged the latency period of inducing platelet plug formation in mesenteric venules. 5 These results indicate that the antiplatelet activity of activated MMP-9 may be involved in the following pathways. (1) Activated MMP-9 may inhibit the activation of phospholipase C, followed by inhibition of phosphoinositide breakdown, protein kinase C activation, and thromboxane A 2 formation, thereby leading to inhibition of intracellular Ca 2+ mobilization. (2) Activated MMP-9 also activated the formation of nitric oxide/cyclic GMP, resulting in inhibition of platelet aggregation. These results strongly indicate that MMP-9 is a potent inhibitor of aggregation. It may play an important role as a negative feedback regulator during platelet activation.

KW - Arterial thrombosis

KW - Immunogold

KW - Matrix metalloproteinase-9

KW - Platelet aggregation

KW - Protein kinase C

UR - http://www.scopus.com/inward/record.url?scp=4644299706&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=4644299706&partnerID=8YFLogxK

U2 - 10.1038/sj.bjp.0705917

DO - 10.1038/sj.bjp.0705917

M3 - Article

C2 - 15289295

AN - SCOPUS:4644299706

VL - 143

SP - 193

EP - 201

JO - British Journal of Pharmacology

JF - British Journal of Pharmacology

SN - 0007-1188

IS - 1

ER -