Hydrogen peroxide alters the physical state and function of the plasma membrane of pulmonary artery endothelial cells
- 1 March 1991
- journal article
- research article
- Published by Wiley in Journal of Cellular Physiology
- Vol. 146 (3) , 362-369
- https://doi.org/10.1002/jcp.1041460305
Abstract
Hydrogen peroxide (H2O2) is an important mediator of acute oxidative injury to vascular endothelium. Because the plasma membrane is the initial site of interaction between endothelial cells and extracellular H2O2 produced by stimulated neutrophils or macrophages, we evaluated the effect of H2O2 on the physical state, i.e., fluidity, and function of porcine pulmonary artery endothelial cell plasma membranes. Lactate dehydrogenase (LDH) release, 5‐hydroxytryptamine (5‐HT) uptake, limiting fluorescence anisotropy (r∞) for trimethylamino‐diphenylhexatriene (TMA‐DPH), and conjugated dienes were measured 0.5, 6, and 24 hr after cells were exposed for 30 min to 50‐μM H2O2 or Hanks' Balanced Salt Solution (control). Compared with control cells, H2O2 caused significant increases in LDH release and in 5‐HT uptake 6 hr after exposure. The increase in 5‐HT uptake was not blocked by imipramine. H2O2 also caused a significant increase in r∞ for TMA‐DPH 0.5 hr after exposure and a significant reduction in r∞ for TMA‐DPH 6 hr after exposure. Cellular contents of conjugated dienes were increased 0.5 and 6 hr after exposure to H2O2. Twenty‐four hours after exposute LDH release, r∞, 5‐HT uptake, and conjugated dienes had returned to control levels. Preincubation with 50‐μM α‐tocopherol (vitamin E) or 1‐mM or 10‐mM dimethylthiourea (DMTU) for 1 hr or 24 hr prevented endothelial cell injury, whereas addition of vitamin E or DMTU to the medium 1 hr or 3 hr after H2O2 exposure did not protect against injury. These results indicate that H2O2 causes significant damage to the plasma membrane of pulmonary artery endothelial cells in vitro, leading to alterations in fluidity and leakiness of the membrane. This injury is associated with membrane lipid peroxidation, is reversible, and can be prevented by pretreatment, but not by post‐treatment, with vitamin E or DMTU.Keywords
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