H2O2 causes endothelial barrier dysfunction without disrupting the arginine-nitric oxide pathway.
We have previously demonstrated that nitric oxide (.NO) donors attenuate and that inhibition of endogenous nitric oxide synthase (NOS) enhances hydrogen peroxide (H2O2)-mediated porcine pulmonary artery endothelial cell (PAEC) injury. The current study investigates the hypothesis that oxidant-mediated inhibition of NOS contributes to PAEC injury. PAEC barrier function, measured as the transmonolayer clearance of albumin, was significantly impaired by H2O2 (10-100 microM) in the absence of cytotoxicity. Treatment with H2O2 did not alter NOS activity, measured as the conversion of [3H]arginine to [3H]citrulline in PAEC lysates, either immediately after treatment with 0-250 microM H2O2 for 30 min or for up to 120 min after treatment with 100 microM H2O2. H2O2 had little effect on NOS activity in intact PAECs, measured as 1) the formation of [3H]citrulline in [3H]arginine-loaded PAECs, 2) PAEC guanosine 3',5'-cyclic monophosphate content, and 3) PAEC.NO release to the culture media. These results indicate that the arginine-.NO pathway remains intact after exposure to oxidant conditions sufficient to promote functional derangements of vascular endothelial cells.
Gupta
M P
MP
Department of Medicine, Indiana University, Indianapolis, USA.
Steinberg
H O
HO
Hart
C M
CM
D000818:Animals; D001120:Arginine / Q000378:metabolism*; D002199:Capillary Permeability / Q000187:drug effects*; D002478:Cells, Cultured; D002956:Citrulline / Q000096:biosynthesis; D006152:Cyclic GMP / Q000378:metabolism; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D009569:Nitric Oxide / Q000378:metabolism*; D019001:Nitric Oxide Synthase / Q000378:metabolism; D016877:Oxidants / Q000494:pharmacology*; D011651:Pulmonary Artery / Q000166:cytology; D013552:Swine
D016877:Oxidants; D002956:Citrulline; D009569:Nitric Oxide; D001120:Arginine; D006861:Hydrogen Peroxide; D019001:Nitric Oxide Synthase; D006152:Cyclic GMP