Biochemistry (Moscow)

, Volume 74, Issue 6, pp 605–612 | Cite as

Activation and damage of endothelial cells upon hypoxia/reoxygenation. Effect of extracellular pH

  • O. A. Antonova
  • S. A. Loktionova
  • Yu. A. Romanov
  • O. N. Shustova
  • M. V. Khachikian
  • A. V. MazurovEmail author


Disturbances of blood flow upon vascular occlusions and spasms result in hypoxia and acidosis, while its subsequent restoration leads to reoxygenation and pH normalization (re-alkalization) in ischemic sites of the vascular bed. The effect of hypoxia/reoxygenation on activation and stimulation of apoptosis in cultured human endothelial cells was studied. The cells were subjected to hypoxia (2% O2, 5% CO2, 93% N2) for 24 h followed by reoxygenation (21% O2, 5% CO2, 74% N2) for 5 h. Reoxygenation was carried out at different pH-6.4 (preservation of acidosis after hypoxia), 7.0, and 7.4 (partial and complete re-alkalization, respectively). Hypoxia only slightly (by ∼30%) increased the cell adhesion molecule ICAM-1 content on the cell surface, whereas reoxygenation more than doubled its expression. The reoxygenation effect depended on the medium acidity, and ICAM-1 increase was more pronounced at pH 7.0 compared to that at pH 6.4 and 7.4. Neither hypoxia nor reoxygenation induced expression of two other cell adhesion molecules, VCAM and E-selectin. Incubation of cells under hypoxic conditions but not reoxygenation stimulated secretion of von Willebrand factor and increased its concentration in the culture medium by more than 4 times. The percentage of cells containing apoptosis marker, activated caspase-3, was increased by approximately 1.5 times upon hypoxia as well as hypoxia/reoxygenation. Maximal values were achieved when reoxygenation was performed at pH 7.0. These data show that hypoxia/reoxygenation stimulate pro-inflammatory activation (ICAM-1 expression) and apoptosis (caspase-3 activation) of endothelial cells, and the extracellular pH influences both processes.

Key words

endothelium hypoxia reoxygenation apoptosis cell adhesion molecules von Willebrand factor extracellular pH 



endothelial cell


intercellular cell adhesion molecule-1


tumor necrosis factor


vascular cell adhesion molecule


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Copyright information

© Pleiades Publishing, Ltd. 2009

Authors and Affiliations

  • O. A. Antonova
    • 1
  • S. A. Loktionova
    • 1
  • Yu. A. Romanov
    • 1
  • O. N. Shustova
    • 1
  • M. V. Khachikian
    • 1
  • A. V. Mazurov
    • 1
    Email author
  1. 1.Russian Cardiology Research and Production ComplexMoscowRussia

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