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Modulation of Endothelial Function by Hypoxia: Perturbation of Barrier and Anticoagulant Function, and Induction of a Novel Factor X Activator

  • Satoshi Ogawa
  • Revati Shreeniwas
  • Caesar Butura
  • Jerold Brett
  • David M. Stern
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 281)

Abstract

Exposure of the vessel wall to hypoxemia is a central feature of ischemic cardiovascular disease. This led us to examine the perturbation of endothelial cell properties under hypoxia. An atmosphere of pO2 of 12 mmHg is not lethal to the endothelial cells for up to five days, but barrier function was impaired. Increased passage of macromolecule tracers were observed in time-and dose-dependent manner and electron microscopy demonstrated small gaps (0.5 — 1.0 µm) between cells. Expression of the anticoagulant cofactor thrombomodulin was also perturbed: thrombomodulin activity and antigen decreased in parallel. Northern blots showed almost complete suppression of thrombomodulin in hypoxic culture. Furthermore, synthesis of other proteins, such as fibronectin, was slightly enhanced under hypoxia. In addition to the suppression of these anticoagulant cofactor, hypoxic endothelial cell displayed a noval procoagulant activity distinct from tissue factor. Further study revealed that hypoxic endothelial cultures directly activated Factor X, as assessed by functional assays and SDS-PAGE. In addition to this no activation of Factor IX or prothrombin was observed. The hypoxia-induced Factor X activator was membrane-associated, required calcium to form Factor Xa, was inhibited by HgCl2 but not by PMSF, and had Km ≈ 25 µg/ml. Co-incubation of hypoxic cultures with cycloheximide prevented the expression of this activity, suggesting that protein synthesis is required for its expression. These functional perturbations of endothelial cells were reversible following reoxygenation. These data indicate that hypoxia imposes a selective perturbation on endothelial cell function, suggesting the possible contribution of hypoxemia to vascular dysfunction in ischemia.

Keywords

Tissue Factor Procoagulant Activity Bovine Aortic Endothelial Cell Endothelial Monolayer Hypoxic Culture 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer Science+Business Media New York 1990

Authors and Affiliations

  • Satoshi Ogawa
    • 1
  • Revati Shreeniwas
    • 1
  • Caesar Butura
    • 1
  • Jerold Brett
    • 1
  • David M. Stern
    • 1
  1. 1.Department of Physiology and Cellular BiophysicsCollege of Physicians and Surgeons of Columbia UniversityNew YorkUSA

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