Induction of KDR expression in bovine arterial endothelial cells by thrombin: Involvement of nitric oxide

Autor: Jie Wang, Mitsue Onodera, Sei-itsu Murota, Ikuo Morita
Rok vydání: 2002
Předmět:
Transcriptional Activation
Vascular Endothelial Growth Factor A
Physiology
Clinical Biochemistry
Down-Regulation
Endothelial Growth Factors
Nitric Oxide
chemistry.chemical_compound
Thrombin
medicine
Animals
Receptors
Growth Factor
RNA
Messenger
Promoter Regions
Genetic
Protein kinase A
Cells
Cultured
Protein Kinase C
Protein kinase C
Lymphokines
Vascular Endothelial Growth Factors
Receptor Protein-Tyrosine Kinases
Kinase insert domain receptor
Arteries
Cell Biology
Molecular biology
Up-Regulation
Vascular endothelial growth factor
Endothelial stem cell
Vascular endothelial growth factor A
Receptors
Vascular Endothelial Growth Factor
chemistry
Cattle
Endothelium
Vascular
Mitogen-Activated Protein Kinases
Nitric Oxide Synthase
Tyrosine kinase
Cell Division
Gene Deletion
circulatory and respiratory physiology
medicine.drug
Zdroj: Journal of Cellular Physiology. 190:238-250
ISSN: 1097-4652
0021-9541
Popis: Thrombin, a multifunctional serine protease, is generated at the site with vascular injuries. It not only participates in the coagulation cascade, but also can induce a lot of events related to cell mitogenesis and migration. In this study, we investigated the effect of thrombin on endothelial cell proliferation induced by vascular endothelial growth factor (VEGF). Thrombin promoted proliferation of cultured bovine carotid endothelial cells in a time- and dose-dependent manner. Moreover, it drastically enhanced the cell growth stimulated by VEGF. This stimulatory effect was reduced by inhibitors of either protein kinase C (PKC) or mitogen-activated protein kinase kinase (MAPKK). Thrombin induced a significant increase in the level of mRNA of the kinase domain-containing receptor (KDR), but not tms-like tyrosine kinase (Flt-1), in a time-dependent manner, which reached the maximum after 24 h of stimulation. This increase coincides well with the KDR protein expression. The luciferase assay showed that thrombin induced an about 7.5-fold increase in the KDR promoter activity compared with the control. This enhanced KDR promoter activity was also abolished by inhibitors of either PKC or MAPKK. The deletion analyses indicated that the region between -115 and -97 (containing Sp1 binding region) within the KDR promoter gene was required for the enhanced KDR expression induced by thrombin and VEGF. Moreover, the nitric oxide synthase (NOS) inhibitor abolished both the accelerated cell proliferation and the increased KDR expression induced by thrombin and VEGF. This inhibition was abrogated by DETA NONOate, a NO donor with long half-life. These findings suggest that thrombin might potentiate the VEGF-induced angiogenic activity through increasing the level of the VEGF receptor KDR, in which production of NO is involved.
Databáze: OpenAIRE
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