thrombin-receptor-peptide-(42-55) and Obesity

During thrombosis, alpha-thrombin becomes sequestered by fibrin and the subendothelial basement membrane, and it is available to interact with the vasculature over prolonged periods. In this study, we investigated the long-term effect of alpha-thrombin on G alpha i3 and G alpha s levels in human vascular endothelial cells (EC). Because obesity is associated with changes in receptor signaling in many animal models, we also explored the influence of this clinical risk factor.. Primary cultures of human EC were exposed to alpha-thrombin for 16 hours, and immunologically detectable G alpha i3 and G alpha s levels were measured.. alpha-Thrombin (100 nmol/L) increased G alpha i3 levels in EC derived from the cerebral microvasculature and superficial temporal artery (4.2 +/- 1.2-fold and 2.8 +/- 0.32-fold, respectively) but had no significant effect on EC derived from omental artery (P > .6) or from the superficial temporal artery of obese (body mass index > or = 28 kg/m2) patients (P > .4). The expression of G alpha s was unchanged in all cell types (P > or = .1). Two other circulating peptides, vasoactive intestinal peptide and endothelin-1, failed to alter the expression of either G protein in EC from the cerebral microvasculature, further demonstrating the specificity of the alpha-thrombin effect. However, thrombin receptor activating protein-14 mimicked the alpha-thrombin response and increased G alpha i3 in EC derived from the cerebral microvasculature and superficial temporal artery.. We conclude that alpha-thrombin increases G alpha i3 expression in some EC through activation of its tethered liganded receptor. Obesity appears to suppress this action of alpha-thrombin.

Topics: Adult; Aged; Aged, 80 and over; Body Mass Index; Cells, Cultured; Cerebral Arteries; Cycloheximide; Endothelium, Vascular; Female; Gene Expression Regulation; GTP-Binding Protein alpha Subunits, Gi-Go; GTP-Binding Protein alpha Subunits, Gs; Hirudins; Humans; Male; Middle Aged; Obesity; Omentum; Peptide Fragments; Protein Synthesis Inhibitors; Receptors, Thrombin; Signal Transduction; Temporal Arteries; Thrombin