Function of cell membranes in cerebral cortical tissue of newborn piglets after hypoxia and inhibition of nitric oxide synthase.
Hypoxia-induced brain cell membrane lipid peroxidation can be caused by free radicals that are produced during hypoxia. Recently, the production of nitric oxide (NO), a free radical, has been shown to be increased during cerebral hypoxia-ischemia. The present study tested the hypothesis that inhibition of NO synthase (NOS) reduced hypoxia-induced modifications of Na+,K+-ATPase activity, lipid peroxidation, and [3H]MK-801 binding to the N-methyl-D-aspartate (NMDA) receptor in cerebral cortical tissue of newborn piglets. Studies were performed in 26 newborn piglets. Cerebral NOS was inhibited by the i.v. administration of 25 or 50 mg/kg N(omega)-nitro-L-arginine (NNLA) over 30 min. Control animals received normal saline. Six groups of piglets were thus created (normoxia, no NNLA; normoxia + NNLA 25 mg/kg; normoxia + NNLA 50 mg/kg; hypoxia, no NNLA; hypoxia + NNLA 25 mg/kg; hypoxia + NNLA 50 mg/kg). One hour after the start of NNLA or saline infusion, hypoxia was induced by lowering the FiO2 to 0.07 in the three hypoxia groups, whereas in the three other groups normoxia was maintained. After 60 min of hypoxia, the brain was taken out and frozen. NOS activity, Na+,K+-ATPase activity, conjugated dienes, and [3H]MK-801 binding to the NMDA receptor of cerebral cortical tissue were determined. NOS activity was reduced to 34% of its baseline value with NNLA 25 mg/kg, and to 19-27% of its baseline value with NNLA 50 mg/kg, respectively. Administration of NNLA did neither significantly alter the hypoxia-induced production of conjugated dienes, indicating lipid peroxidation nor the decrease of Na+,K+-ATPase activity after hypoxia. [3H]MK-801 binding studies of the NMDA receptor, however, showed that NNLA preserved Bmax and Kd after hypoxia. We conclude that inhibition of NOS does not change the hypoxia-induced decrease of Na+,K+-ATPase activity and production of conjugated dienes in brain cell membranes. Inhibition of NOS preserved the binding of [3H]MK-801 to the NMDA receptor after hypoxia.
Groenendaal
F
F
Department of Physiology, University of Pennsylvania School of Medicine, Philadelphia 19104, USA.
Mishra
O P
OP
McGowan
J E
JE
Hoffman
D J
DJ
Delivoria-Papadopoulos
M
M
D000818:Animals; D000831:Animals, Newborn; D002462:Cell Membrane / Q000187:drug effects; D002540:Cerebral Cortex / Q000187:drug effects*; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D002534:Hypoxia, Brain / Q000188:drug therapy* / Q000503:physiopathology; D015227:Lipid Peroxidation / Q000187:drug effects*; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors*; D019335:Nitroarginine / Q000494:pharmacology; D011869:Radioligand Assay; D016194:Receptors, N-Methyl-D-Aspartate / Q000378:metabolism; D000254:Sodium-Potassium-Exchanging ATPase / Q000037:antagonists & inhibitors; D013552:Swine
D004791:Enzyme Inhibitors; D016194:Receptors, N-Methyl-D-Aspartate; D019335:Nitroarginine; D019001:Nitric Oxide Synthase; D000254:Sodium-Potassium-Exchanging ATPase