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(Circulation. 2003;107:1520.)
© 2003 American Heart Association, Inc.

Basic Science Reports

L-4F, an Apolipoprotein A-1 Mimetic, Restores Nitric Oxide and Superoxide Anion Balance in Low-Density Lipoprotein-Treated Endothelial Cells

Zhijun Ou, MD; Jingsong Ou, MD, PhD^*; Allan W. Ackerman, MS; Keith T. Oldham, MD; Kirkwood A. Pritchard, Jr, PhD^*

From the Department of Surgery (Z.O., J.O., A.W.A., K.T.O., K.A.P.), the Division of Pediatric Surgery and Pharmacology & Toxicology (K.A.P.), and the Cardiovascular Center (Z.O., J.O., K.T.O., K.A.P.) and Free Radical Research Center (Z.O., J.O., K.T.O., K.A.P.), Medical College of Wisconsin, Children’s Hospital of Wisconsin, Milwaukee.

Correspondence to Kirkwood A. Pritchard Jr, PhD, Medical College of Wisconsin, 8701 Watertown Plank Rd, Milwaukee, WI 53226. E-mail kpritch{at}mcw.edu

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.

Methods and 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.

Key Words: apolipoproteins • cells, endothelial • lipoproteins • nitric oxide • nitric oxide synthase

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