L-4F, an apolipoprotein A-1 mimetic, restores nitric oxide and superoxide anion balance in low-density lipoprotein-treated endothelial cells.
BACKGROUND
Low-density lipoprotein (LDL) impairs endothelial cell function by uncoupling endothelial nitric oxide synthase (eNOS) activity, which allows superoxide anion (O2*-)) to be generated rather than nitric oxide (*NO). Recent reports indicate that apolipoprotein (apo) A-1 mimetics inhibit the development of atherosclerotic lesions in LDL receptor-null mice. Here we hypothesize that L-4F, an apoA-1 mimetic that inhibits atherosclerosis induced by hypercholesterolemia, protects endothelial cell function by preventing LDL from uncoupling eNOS activity.


RESULTS
Bovine aortic endothelial cells were incubated with LDL+/-L-4F, and changes in A23187-stimulated.NO and O2*- generation were determined by ozone chemiluminescence and superoxide dismutase-inhibitable ferricytochrome c reduction, respectively. Western analysis of eNOS immunoprecipitates was used to determine effects of LDL and L-4F on heat shock protein 90 (hsp90) interactions with eNOS. LDL decreased.NO production and increased eNOS-dependent O2*- generation. Pretreatment of LDL with L-4F increased.NO and decreased O2*- generation. By itself, L-4F had no effect on O2*- but did increase *NO generation. Stimulation of endothelial cells incubated with LDL decreased the association of hsp90 with eNOS. Pretreatment of LDL with L-4F prevented a decrease in hsp90 association with eNOS and often enhanced association on stimulation.


CONCLUSIONS
These data demonstrate that L-4F protects endothelial cell function by preventing LDL from uncoupling eNOS activity. L-4F allows endothelial cell to maintain coupled eNOS activity to generate.NO even in the face of atherogenic concentrations of LDL.
Ou
Zhijun
Z
Department of Surgery, Medical College of Wisconsin, Children's Hospital of Wisconsin, Milwaukee 53226, USA.
Ou
Jingsong
J
Ackerman
Allan W
AW
Oldham
Keith T
KT
Pritchard
Kirkwood A
KA
D000818:Animals; D016632:Apolipoprotein A-I; D002417:Cattle; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D018841:HSP90 Heat-Shock Proteins / Q000378:metabolism; D008077:Lipoproteins, LDL / Q000037:antagonists & inhibitors*; D009569:Nitric Oxide / Q000378:metabolism*; D019001:Nitric Oxide Synthase / Q000378:metabolism; D052250:Nitric Oxide Synthase Type III; D010455:Peptides / Q000737:chemistry / Q000494:pharmacology*; D013481:Superoxides / Q000378:metabolism*
D016632:Apolipoprotein A-I; D018841:HSP90 Heat-Shock Proteins; C472885:L-4F peptide; D008077:Lipoproteins, LDL; D010455:Peptides; D013481:Superoxides; D009569:Nitric Oxide; D019001:Nitric Oxide Synthase; D052250:Nitric Oxide Synthase Type III