title: Enhanced secretion of tissue plasminogen activator by simultaneous use of retinoic acid and ascorbic acid from tissue cultured gastroepiploic artery.
abstract: Tissue-type plasminogen activator (tPA) is a key enzyme in the fibrinolysis system and the regulation of its expression has been extensively studied in cultured vascular endothelial cells. Many kinds of supplements including growth factors are needed, however, to keep endothelial cells viable, which leads the culture condition far from the physiological milieu. Using a new device of amorphous calcium phosphate coated culture plate, we succeeded in culturing ring-cut gastroepiploic artery in a basic medium of RPMI 1640 containing 10% fetal calf serum. The overall normal vessel architecture and the antigenicity of von Willebrand factor, tPA and plasminogen activator inhibitor type 1 (PAI-1) were retained for at least 9 days. tPA was constantly secreted into the conditioned medium at least up to day 12. Employing this organ culture technique, we analyzed the effects of two well-known profibrinolytic vitamins of retinoic acid (Vit. A) and ascorbic acid (Vit. C) on the release of tPA and PAI-1. The cultured artery responded well and the tPA secretion was enhanced by factors of 1.5 fold by Vit. A, 1.7 fold by Vit C and 3.2 fold by their combination, whereas none of these stimuli increased PAI-1 secretion. These results suggested that the cultured ring-cut artery retained functional endothelial cells for at least 9 days and was suitable in analyzing the regulatory mechanism of protein synthesis and secretion from the vascular wall. Using this method, vitamins A and C were shown to lead the intravascular condition to a profibrinolytic state.
authors.affiliation: Emergency Unit, Hamamatsu University School of Medicine, Japan. yoshino@hmedc.or.jp
authors.forename: Atsuto
authors.initials: A
authors.lastname: Yoshino
authors.forename: Kazuya
authors.initials: K
authors.lastname: Suzuki
authors.forename: Tetsumei
authors.initials: T
authors.lastname: Urano
authors.forename: Katsunori
authors.initials: K
authors.lastname: Aoki
authors.forename: Yumiko
authors.initials: Y
authors.lastname: Takada
authors.forename: Teruhisa
authors.initials: T
authors.lastname: Kazui
authors.forename: Akikazu
authors.initials: A
authors.lastname: Takada
mesh_terms: D001205:Ascorbic Acid / Q000494:pharmacology*; D017077:Culture Media, Conditioned / Q000737:chemistry / Q000378:metabolism; D004338:Drug Combinations; D004730:Endothelium, Vascular / Q000187:drug effects* / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D024405:Gastroepiploic Artery / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D007231:Infant, Newborn; D009924:Organ Culture Techniques / Q000379:methods; D017395:Plasminogen Activator Inhibitor 1 / Q000378:metabolism; D010959:Tissue Plasminogen Activator / Q000378:metabolism*; D014212:Tretinoin / Q000494:pharmacology*; D014471:Umbilical Veins / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D014841:von Willebrand Factor / Q000378:metabolism
chemical_list: D017077:Culture Media, Conditioned; D004338:Drug Combinations; D017395:Plasminogen Activator Inhibitor 1; D014841:von Willebrand Factor; D014212:Tretinoin; D010959:Tissue Plasminogen Activator; D001205:Ascorbic Acid