Nishida K, Miyazawa Y, Hatano M, Suzuki K, Hirose A, Fukushima R, Okinaga K
Second Department of Surgery, Teikyo University School of Medicine, Itabashi-ku, Tokyo, Kaga, 2-11-1, Japan.
J Surg Res. 1998 Sep;79(1):85-90. doi: 10.1006/jsre.1998.5390.
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.
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.
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.
The present study suggests that 120 min of hypoxia/reoxygenation induces endothelial dysfunction of ECs but does not cause cell death.
内皮细胞在调节血栓形成和止血过程中起关键作用。在本研究中，我们试图在体外对缺氧/复氧后的内皮功能障碍和内皮细胞损伤进行特征描述。
将培养的人脐静脉内皮细胞（ECs）暴露于120分钟的缺氧环境后再进行复氧。检测血栓调节蛋白（TM）的释放和前列腺素I2（PGI2）的生成。通过两种检测方法，即Fura-2释放检测法和51铬（51Cr）释放检测法，来测定缺氧/复氧过程中的内皮细胞损伤情况。
在常氧孵育期间，未检测到ECs释放TM，而在缺氧孵育期间TM释放略有增加。复氧后，TM的释放增加，与缺氧孵育相比，复氧120分钟时TM释放显著更高。在缺氧孵育期间，PGI2的生成显著减少，复氧后30分钟内进一步减少，但在复氧120分钟时恢复到常氧水平。在Fura-2释放检测中，与缺氧孵育相比，在缺氧/复氧过程中观察到Fura-2快速且显著更多地释放。在显示细胞死亡的51Cr释放检测中，缺氧/复氧过程中51Cr释放未增加。
本研究表明，120分钟的缺氧/复氧诱导了ECs的内皮功能障碍，但未导致细胞死亡。
