Assessment of production of endothelium-derived relaxing factor (EDRF) by cultured human vascular endothelial cells based on its anti-aggregatory effect on human platelets.
In order to establish an experimental system to assess the production of endothelium-derived relaxing factor (EDRF) by cultured human umbilical vascular endothelial cells (HUVECs), the effect of endothelial cells on thrombin-induced platelet aggregation was examined. Cultured HUVECs were harvested from umbilical veins by collagenase treatment. The platelet aggregation experiments were performed using cuvettes lined with HUVECs. The cuvettes were prepared by seeding HUVECs in gelatin-coated cuvettes at a cell density of 2 x 10(5) cells/ml and culturing for 48 hours. Thrombin-induced platelet aggregation was inhibited in the presence of HUVECs. This HUVEC-dependent anti-platelet aggregatory effect was enhanced by the addition of bradykinin, which stimulates the production of EDRF, and thrombin-induced platelet aggregation was completely inhibited. Indomethacin reduced the HUVEC-dependent anti-platelet aggregatory effect. These findings suggest that this simple, new experimental system is useful in assessing the production of EDRF by HUVECs and in examining the effects of various chemicals (or agents) on EDRF production.
Ohno
M
M
Department of Dermatology, Shimane Medical University.
Kishimoto
T
T
Jidoi
J
J
Tada
M
M
D001681:Biological Assay / Q000379:methods; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000378:metabolism*; D006801:Humans; D009569:Nitric Oxide / Q000096:biosynthesis*; D010974:Platelet Aggregation
D009569:Nitric Oxide