The effect of thrombin on low-density lipoprotein permeability and uptake by an arterial endothelial smooth muscle cell bilayer.
Thrombin, a mediator of thrombosis, has been shown to directly alter the function of vascular cells. We studied the effect of thrombin on low-density lipoprotein permeability and uptake by an arterial endothelial cell-smooth muscle cell bilayer to determine its potential role in atherogenesis. Confluent cell bilayers were incubated in media containing thrombin (10 or 50 units/ml) for a period of 24 hours to 9 days. Iodine 125 (125I)-LDL (10 micrograms protein/ml) was then added to the media, and after a 3-hour incubation, 125I-LDL transit through the endothelial cell layer as well as membrane binding and uptake by each cell type were measured. The lower concentration of thrombin caused a delayed increase in both the permeability (p less than 0.0001) and uptake (p less than 0.05) of LDL, but had no effect on membrane binding of the lipoprotein. The higher thrombin concentration led to an immediate increase in endothelial cell permeability to LDL (p less than 10(-7)) and a significant reduction in both cellular uptake (p less than 10(-7)) and membrane binding (p less than 0.0005). In contrast, smooth muscle cell binding and uptake were unaffected at the lower concentration of thrombin. At the higher concentration, smooth muscle cell uptake of LDL was increased (p less than 10(-7)) disproportionately to a more limited increase in membrane binding (p less than 0.05). Endothelial DNA content, reflecting cell number, was increased at 10 units/ml thrombin (p less than 0.001) but markedly reduced at 50 units/ml thrombin (p less than 0.0005), whereas smooth muscle cell DNA content remained unchanged.(ABSTRACT TRUNCATED AT 250 WORDS)
Alexander
J J
JJ
Department of Surgery, Case Western Reserve University, Cleveland Metropolitan General Hospital, OH 44109.
Miguel
R
R
Graham
D
D
Piotrowski
J J
JJ
D000818:Animals; D001011:Aorta / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D002417:Cattle; D002452:Cell Count / Q000187:drug effects; D002463:Cell Membrane Permeability / Q000187:drug effects*; D002478:Cells, Cultured; D004247:DNA / Q000032:analysis; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D007457:Iodine Radioisotopes; D008077:Lipoproteins, LDL / Q000378:metabolism*; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D013917:Thrombin / Q000494:pharmacology*
D007457:Iodine Radioisotopes; D008077:Lipoproteins, LDL; D004247:DNA; D013917:Thrombin