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Correspondence to:
Santhini Ramasamy.
Hypercholesterolemia is associated with impairments in endothelium-dependent vascular relaxations. Paradoxically, endothelial production of nitrogen oxides is increased in early stages of hypercholesterolemia. Prior work has shown that oxidized low density lipoprotein (LDL) has both stimulatory and inhibitory effects on endothelial nitric oxide synthase expression (eNOS) and has focused on lysophosphatidyl choline (LPC) as a component of oxidized LDL which may modulate this effect. Another biologically active component of oxidized LDL is 13-hydroperoxyoctadecadienoic acid (13-HPODE), an oxidized form of linoleic acid. The purpose of this study was to determine the effect of HPODE on the expression of eNOS in bovine aortic endothelial cells (BAECs). Twenty four hour treatment of endothelial cells with HPODE caused a dose-dependent increase in eNOS mRNA levels as assessed by Northern analysis. The time response studies show that HPODE treatment significantly increased eNOS mRNA levels at 12 and 24 h. Concomitant with the increase in eNOS mRNA levels, 20 µM HPODE treatment significantly increased eNOS protein content and enzyme activity. Nuclear run-on studies indicated that the rate of transcription of eNOS gene was significantly elevated 4 h after HPODE treatment when compared to control cultures. Also, actinomycin D studies demonstrated that the half-life of eNOS mRNA was increased from 6 h to 12 h by HPODE treatment. Thus, HPODE-induced up-regulation of eNOS expression is mediated by both transcriptional and posttranscriptional mechanisms.
These observations suggest that endothelial cells may attempt to compensate for oxidative injury by increasing expression of eNOS in early stages of hypercholesterolemia.—Ramasamy, S., S. Parthasarathy, and D. G. Harrison. Regulation of endothelial nitric oxide synthase gene expression by oxidized linoleic acid. J. Lipid Res. 1998. 39: 268–276.
Supplementary key words:
HPODE, endothelial cells, nitric oxide synthase, nuclear run-on assay, eNOS mRNA stability
Copyright © 1998 by Lipid Research, Inc.
Original Article
Regulation of endothelial nitric oxide synthase gene expression by oxidized linoleic acid
Santhini Ramasamya,
Sampath Parthasarathyb, and
David G. Harrisona
a Division of Cardiology, Department of Medicine, Department of Obstetrics and Gynecology, 1639 Pierce Drive, Atlanta, GA 30322
b Division of Cardiology, Department of Medicine, Emory University School of Medicine, 1639 Pierce Drive, Atlanta, GA 30322
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