Reperfusion induces sublethal endothelial injury.
BACKGROUND
Endothelial cells are pivotal in regulating thrombosis and hemostasis. In this study, we sought to characterize endothelial dysfunction and endothelial cell injury in vitro after hypoxia/reoxygenation.


METHODS
Cultured human umbilical vein endothelial cells (ECs) were exposed to 120 min of hypoxia followed by reoxygenation. The release of thrombomodulin (TM) and the production of prostaglandin I2 (PGI2) were measured. Endothelial cell injury in hypoxia/reoxygenation was measured by two assays, the Fura-2 release assay and the 51chromium (51Cr) release assay.


RESULTS
TM release from ECs during normoxic incubation was undetectable, while it was slightly increased during hypoxic incubation. After reoxygenation, the release of TM increased, and it became significantly higher at 120 min after reoxygenation compared with hypoxic incubation. The production of PGI2 significantly decreased during hypoxic incubation and further decreased within 30 min after reoxygenation, but returned to normoxic levels at 120 min after reoxygenation. In the Fura-2 release assay, a rapid and significantly greater release of Fura-2 was observed in hypoxia/reoxygenation compared with hypoxic incubation. In the 51Cr release assay which demonstrates cell death, 51Cr release did not increase in hypoxia/reoxygenation.


CONCLUSIONS
The present study suggests that 120 min of hypoxia/reoxygenation induces endothelial dysfunction of ECs but does not cause cell death.
Nishida
K
K
Second Department of Surgery, Teikyo University School of Medicine, Itabashi-ku, Tokyo, Kaga, 2-11-1, Japan.
Miyazawa
Y
Y
Hatano
M
M
Suzuki
K
K
Hirose
A
A
Fukushima
R
R
Okinaga
K
K
D016923:Cell Death; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000473:pathology / Q000503:physiopathology*; D011464:Epoprostenol / Q000096:biosynthesis; D006801:Humans; D015427:Reperfusion Injury / Q000503:physiopathology*; D018180:Thrombomodulin / Q000096:biosynthesis
D018180:Thrombomodulin; D011464:Epoprostenol