title: Inhibition of thrombin-induced vascular endothelial growth factor production in human neuroblastoma (NB-1) cells by argatroban.
abstract: Thrombin, a serine protease that plays a pivotal role in blood coagulation, wound healing, and angiogenesis, has also been implicated in the mitogenesis of various cell types. Previously, we showed that thrombin and the thrombin receptor agonist peptide (TRAP-14; SFLLRNPNDKYEPF) for protease-activated receptor 1 (PAR1) induce vascular endothelial growth factor (VEGF) secretion in PC-12 cells. In this study, we show that thrombin and TRAP-14 also stimulate VEGF secretion in the human NB-1 neuroblastoma cells. In these cells, we further show that thrombin-induced VEGF secretion was blocked by cycloheximide and actinomycin D, indicating that de novo protein synthesis is essential for this process. Reduced thrombin-induced VEGF secretion upon treatment with LY294002, calphostin C, or BAPTA, further suggests that the process is dependent on phosphatidyl-inositol-3-kinase, protein kinase C, and calcium. However, the complete loss of thrombin-induced VEGF production upon treatment with argatroban, a derivative of arginine and a potent anticoagulant/antithrombin agent, supports the notion that argatroban serves as a useful therapeutic tool for thrombin-associated pathologic conditions. Here, it appears that argatroban may be effective in controlling disorders linked to thrombin-induced VEGF production in neuronal cells.
authors.affiliation: Department of Laboratory and Vascular Medicine, Cardiovascular and Respiratory Disorders, Graduate School of Medical and Dental Sciences, Kagoshima University, Kagoshima, Japan. kpsarker@ucalgary.ca
authors.forename: Krishna Pada
authors.initials: KP
authors.lastname: Sarker
authors.forename: Kamal Krishna
authors.initials: KK
authors.lastname: Biswas
authors.forename: Kazuyo
authors.initials: K
authors.lastname: Yamaji
authors.forename: Munekazu
authors.initials: M
authors.lastname: Yamakuchi
authors.forename: Teruto
authors.initials: T
authors.lastname: Hashiguchi
authors.forename: Ki-Young
authors.initials: KY
authors.lastname: Lee
authors.forename: Ikuro
authors.initials: I
authors.lastname: Maruyama
mesh_terms: D000818:Animals; D001120:Arginine / Q000031:analogs & derivatives; D002118:Calcium / Q000378:metabolism; D020013:Calcium Signaling / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006490:Hemostatics / Q000494:pharmacology*; D006801:Humans; D009447:Neuroblastoma / Q000378:metabolism*; D016716:PC12 Cells; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D010875:Pipecolic Acids / Q000494:pharmacology*; D010975:Platelet Aggregation Inhibitors / Q000494:pharmacology*; D011493:Protein Kinase C / Q000378:metabolism; D051381:Rats; D013449:Sulfonamides; D013917:Thrombin / Q000494:pharmacology*; D042461:Vascular Endothelial Growth Factor A / Q000096:biosynthesis*
chemical_list: D001120:Arginine; D002118:Calcium; D004791:Enzyme Inhibitors; D006490:Hemostatics; D019869:Phosphatidylinositol 3-Kinases; D010875:Pipecolic Acids; D010975:Platelet Aggregation Inhibitors; D011493:Protein Kinase C; D013449:Sulfonamides; D013917:Thrombin; D042461:Vascular Endothelial Growth Factor A; C031942:argatroban