Mechanisms involved in the contraction of endothelial cells by hydrogen peroxide.
The importance of endothelial contraction in the genesis of inflammatory edema has been reported. ROS are metabolites synthesized in pathological conditions in that a significant intravascular fluid leak occurs, such as ischemia-reperfusion. Present experiments were designed to test the hypothesis that ROS, particularly H2O2, may elicit the contraction of endothelial cells, and to explore the mechanisms involved. Bovine aortic endothelial cells incubated with H2O2 showed a significant reduction in planar cell surface area (PCSA), and a significant increase in myosin light chain phosphorylation (MLCP), with a time- and dose-dependent pattern, without any significant toxicity. This effect of H2O2 was not blocked by sulotroban (TxA2 antagonist) or BN 52021 (PAF antagonist). Lanthanum chloride (calcium channel blocker) and EGTA partially inhibited the increase in MLCP induced by H2O2. H7 and staurosporine, PKC inhibitors, and PKC down-regulation (phorbol myristate acetate treatment, 24 h) also blocked H2O2-dependent endothelial contraction, measured as PCSA or MLCP. H2O2 increased the intracellular calcium concentration, an effect blunted by EGTA and lanthanum chloride. H2O2 also increased the phosphorylation of an 80 kD polypeptide, probably MARCKS, a PKC substrate. In summary, the present results demonstrate the ROS-dependent contraction of endothelial cells, an effect that could explain the intravascular fluid leak observed in some pathophysiological situations. Calcium and PKC may be involved in the development of this contraction.
López-Ongil
S
S
Department of Physiology and Pharmacology, Alcalá de Henares University, Madrid, Spain.
Torrecillas
G
G
Pérez-Sala
D
D
González-Santiago
L
L
Rodríguez-Puyol
M
M
Rodríguez-Puyol
D
D
D000818:Animals; D001011:Aorta; D002417:Cattle; D048429:Cell Size / Q000187:drug effects; D002478:Cells, Cultured; D006152:Cyclic GMP / Q000378:metabolism; D004224:Diterpenes; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D016166:Free Radical Scavengers / Q000494:pharmacology; D046934:Ginkgolides; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D007700:Kinetics; D007783:Lactones / Q000494:pharmacology; D018994:Myosin Light Chains / Q000378:metabolism; D010766:Phosphorylation; D017382:Reactive Oxygen Species; D013449:Sulfonamides / Q000494:pharmacology
D004224:Diterpenes; D016166:Free Radical Scavengers; D046934:Ginkgolides; D007783:Lactones; D018994:Myosin Light Chains; D017382:Reactive Oxygen Species; D013449:Sulfonamides; C040624:sulotroban; D006861:Hydrogen Peroxide; C045856:ginkgolide B; D006152:Cyclic GMP