Human recombinant interleukin-1 beta- and tumor necrosis factor alpha-mediated suppression of heparin-like compounds on cultured porcine aortic endothelial cells.
Cytokines are known to tip the balance of the coagulant-anticoagulant molecules on the endothelial cell surface toward intravascular coagulation. Their effects on endothelial cell surface-associated heparin-like compounds have not been examined yet. Incorporation of [35S]sulfate into heparan sulfate on cultured porcine aortic endothelial cells was suppressed by human recombinant interleukin-1 beta (rIL-1 beta) or tumor necrosis factor alpha (rTNF alpha) in a dose- and time-dependent manner with little effect on cell number, protein content, and [3H]leucine incorporation of cells. Maximal inhibition was achieved by incubation of cells with 100 ng/ml of rIL-1 beta or 5 ng/ml of rTNF alpha for 12-24 hours, resulting in a reduction of the synthesis of heparan sulfate on the cell surface by approximately 50%. The dose dependency was consistent with that seen in the stimulation of endothelial cell procoagulant activity by each cytokine. The suppression of heparan sulfate synthesis was sustained for at least 48 hours after pretreatment of cells with cytokines and was unchanged after the addition of indomethacin or polymyxin B. The rate of degradation of prelabeled 35S-heparan sulfate on the cell surface was not altered by cytokine treatments. Neither the size, the net negative charge, nor the proportion of the molecule with high affinity for antithrombin III of endothelial cell heparan sulfate was changed by cytokines. Furthermore, specific binding of 125I-labeled antithrombin III to the endothelial cell surface was reduced to 40-60% of control by cytokines. In parallel with reduction in binding, antithrombin III cofactor (heparin-like) activity was partially diminished in cytokine-treated endothelial cells. Thus, cytokine-mediated suppression of heparin-like substance on endothelial cells appears to be another cytokine-inducible endothelial effects affecting coagulation.
Department of Medicine and Geriatrics, Kochi Medical School, Japan.
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Kobayashi
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Shimada
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Ozawa
D000818:Animals; D000990:Antithrombin III / Q000378:metabolism; D001011:Aorta / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D001685:Biological Factors / Q000494:pharmacology; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D002478:Cells, Cultured; D016207:Cytokines; D004730:Endothelium, Vascular / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D006025:Glycosaminoglycans / Q000096:biosynthesis; D006493:Heparin / Q000235:genetics / Q000378:metabolism*; D007375:Interleukin-1 / Q000502:physiology*; D011485:Protein Binding / Q000187:drug effects; D011994:Recombinant Proteins / Q000502:physiology; D013462:Sulfur Radioisotopes; D013489:Suppression, Genetic / Q000187:drug effects*; D013552:Swine; D014409:Tumor Necrosis Factor-alpha / Q000502:physiology*
D001685:Biological Factors; D016207:Cytokines; D006025:Glycosaminoglycans; D007375:Interleukin-1; D011994:Recombinant Proteins; D013462:Sulfur Radioisotopes; D014409:Tumor Necrosis Factor-alpha; D000990:Antithrombin III; D006493:Heparin