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eNOS Inhibition is Protective against Oxygen Glucose Deprivation in Brain Microvascular Endothelial Cells by Preempting eNOS Uncoupling
Author(s) -
Katz Paige S,
Wappler Edina A,
Katakam Prasad V,
Busija David W
Publication year - 2012
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.26.1_supplement.1058.5
Subject(s) - enos , reactive oxygen species , nitric oxide , endothelial nos , chemistry , endothelial stem cell , medicine , microbiology and biotechnology , endocrinology , nitric oxide synthase , biochemistry , pharmacology , biology , in vitro
Reactive oxygen species (ROS) have been implicated in cellular injury due to oxygen glucose deprivation (OGD). Our objective was to determine whether uncoupling of endothelial nitric oxide synthase (eNOS) contributes to cell injury in endothelial cells exposed to OGD. Brain microvascular endothelial cells from rat (RBMEC) and human (HBMEC) were cultured and exposed either to 2 hours of OGD or normoxia ± N ω‐nitro‐1‐arginine methyl ester (NAME). RBMEC and HBMEC viabilities were reduced by OGD and rescued by NAME dose‐dependently. Low temperature native PAGE revealed reduced eNOS monomer/dimer ratio in RBMEC following OGD compared to control. Fluorescence measurements of dihydroethidium (DHE), a ROS‐sensitive fluoroprobe, displayed increased ROS generation in RBMEC following OGD compared to control that was abolished by NAME. In conclusion, eNOS inhibition prevented ROS generation via eNOS uncoupling and afforded protection against OGD in endothelial cells.