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Signal transduction pathways of bacterial lipopolysaccharide-stimulated bovine vascular endothelial cells

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Abstract

Increased procoagulant activity of vascular endothelial cells may be an important component in the pathogenesis of intravascular coagulation associated with gram-negative bacterial diseases. Two bovine endothelial cell (BEC) lines isolated from pulmonary arteries (ENS-2 and ENT-18) were used in this study to investigate procoagulant signal transduction pathways of endotoxin (lipopolysaccharide, LPS) —stimulated BECs. The endothelial cell line ENS-2 was sensitive to LPS as demonstrated by tissue factor (TF) expression, but in contrast, the ENT-18 endothelial cell line was unusually resistant to the effects of LPS. No remarkable quantitative difference in binding of radiolabeled LPS was detected between the two endothelial cell lines. A protein kinase C (PKC) activator (phorbol 12-myristate 13-acetate, PMA) failed to induce TF expression in either cell line at concentrations ranging from 0.05 to 1.00μM when used as a sole stimulus for the endothelial cells. However, when PMA was used in combination with LPS, PMA enhanced the stimulatory effect of LPS on the endothelial cells. In parallel experiments, PKC inhibitors (H-7 and GF 109203X) interfered with the stimulatory effect of LPS on the cells by decreasing tissue factor expression. We also found that an activator of adenylate cyclase, forskolin, similarly inhibited LPS-induced tissue factor activity. In contrast, protein tyrosine kinase inhibitors (genistein, lavendustin A) had no inhibitory effect on LPS-induced endothelial cell tissue factor expression. Our results collectively suggest that activation of PKC is an important step in stimulation of endothelial cells by LPS, and that LPS and phorbol esters may synergize to produce an enhanced stimulatory effect. Our results also suggest participation of cAMP in controlling LPS-mediated stimulation of endothelial cells, but fail to demonstrate a role for protein tyrosine kinase activity.

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Authors and Affiliations

  1. Department of Pathobiology, The University of Tennessee, Knoxville

    Zhengang Yang, Philip N. Bochsler, Candace D. Carter & Lajwanti S. Khemlani

  2. Department of Medical Biology, The University of Tennessee Medical Center, Knoxville, Tennessee

    Roger C. Carroll

  3. Parke-Davis Pharmaceutical Research, Ann Arbor, Michigan

    Michael A. Breider

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  1. Zhengang Yang
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  6. Michael A. Breider
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Yang, Z., Bochsler, P.N., Carroll, R.C. et al. Signal transduction pathways of bacterial lipopolysaccharide-stimulated bovine vascular endothelial cells. Inflammation 18, 221–233 (1994). https://doi.org/10.1007/BF01534563

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