title: Endothelial cells exhibit differential chemokinetic and mitogenic responsiveness to alpha-thrombin.
abstract: Rapid regeneration of the endothelium is a critical component of vascular wall repair because limitations of this process enhance early thrombotic and vasospastic complications, as well as late sequelae of recurrent lesion formation. We have postulated that direct activation of the thrombin receptor initiates both mitogenic and chemokinetic endothelial behavior which facilitates intimal repair. To characterize the role of the thrombin receptor in human endothelial cell (EC) proliferation and migration, we investigated the effects of both alpha-thrombin (0.5-10 U/ml) and its receptor-activating peptide (TRAP; 1-100 microM). Responses of human aortic (HAEC) and umbilical vein (HUVEC) were characterized using [3H]thymidine and 61Cr microcarrier bead assays of proliferation and migration, respectively. Expression of motility-related genes was evaluated using a ribonuclease protection assay. Thrombin exerts both of its chemokinetic and mitogenic effects differentially in human endothelial cells. Following 2 or 4 days in culture, HUVEC proliferation increased two- to threefold after exposure to thrombin, primarily in the low concentration range (P < 0.05). However, HACE proliferation was inhibited up to 50% after a 4-day incubation period (P < 0.005). These mitogenic effects, including the inhibition of aortic endothelial cell proliferation, were reproduced, in part, by thrombin receptor activation with TRAP. In contrast, thrombin stimulates migratory responses in HAEC, but not HUVEC. However, this behavior was not reproduced by TRAP. It is noteworthy that urokinase-plasminogen activator (u-PA) expression was much more strongly expressed in migrating HAEC than in the HUVEC population. Moreover, when stimulated with thrombin, u-PA gene expression was significantly augmented in HAEC. It has been speculated that an effective human thrombin receptor (HTR) antagonist may reduce the proliferation of vascular smooth muscle cells and the development of a restenotic lesion following arterial wall injury. Our data suggest that such an inhibitor will likely also accelerate intimal regeneration through a dominant effect on limiting the HTR inhibitory effect on endothelial proliferation.
authors.affiliation: Division of General Vascular Surgery, Emory University School of Medicine, Atlanta, Georgia 30322, USA.
authors.forename: H S
authors.initials: HS
authors.lastname: Wang
authors.forename: F
authors.initials: F
authors.lastname: Li
authors.forename: M S
authors.initials: MS
authors.lastname: Runge
authors.forename: E L
authors.initials: EL
authors.lastname: Chaikof
mesh_terms: D000595:Amino Acid Sequence; D001011:Aorta; D002455:Cell Division / Q000187:drug effects*; D002633:Chemotaxis / Q000187:drug effects*; D004730:Endothelium, Vascular / Q000166:cytology* / Q000502:physiology; D005347:Fibroblasts; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D009842:Oligopeptides / Q000494:pharmacology; D011956:Receptors, Cell Surface / Q000235:genetics; D018179:Receptors, Thrombin / Q000235:genetics / Q000502:physiology; D055293:Receptors, Urokinase Plasminogen Activator; D013917:Thrombin / Q000494:pharmacology*; D010959:Tissue Plasminogen Activator / Q000235:genetics; D014471:Umbilical Veins; D014568:Urokinase-Type Plasminogen Activator / Q000235:genetics
chemical_list: D009842:Oligopeptides; C524898:PLAUR protein, human; D011956:Receptors, Cell Surface; D018179:Receptors, Thrombin; D055293:Receptors, Urokinase Plasminogen Activator; C096900:thrombin receptor-activating peptide SFLLRNPNDKY; D013917:Thrombin; D010959:Tissue Plasminogen Activator; D014568:Urokinase-Type Plasminogen Activator