(Circulation. 1996;94:1682-1689.)
© 1996 American Heart Association, Inc.
Articles |
Fluid Flow Inhibits Endothelial Adhesiveness
Nitric Oxide and Transcriptional Regulation of VCAM-1
the Falk Cardiovascular Research Center, Stanford, Calif.
Correspondence to John P. Cooke, MD, PhD, Falk Cardiovascular Research Center, 300 Pasteur Dr, Stanford, CA 94305-5246.
Background In the arterial tree, regions exposed to reduced shear stress (low and/or disturbed flow) are predisposed to atherogenesis. Fluid flow is a potent stimulus for the release of endothelium-derived nitric oxide (NO). Because NO inhibits monocyte–endothelial cell interaction, we speculated that the effects of flow in inhibiting atherogenesis might be mediated in part by NO.
Methods and Results Confluent monolayers of human aortic endothelial cells were exposed to static or fluid flow conditions for 4 hours. The medium was replaced, and cells were then incubated with native LDL (50 µg/mL), oxidized LDL (30 µg/mL), or lipopolysaccharide (LPS) (10 ng/mL) + tumor necrosis factor-
(TNF-) (10 U/mL) for an additional 4 hours. Functional binding assays using THP-1 monocytes were then performed. Superoxide production by human aortic endothelial cells was monitored by lucigenin chemiluminescence, and expression of the adhesion molecules vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 were quantified by flow cytometry. Whereas native LDL had little effect, incubation with either oxidized LDL or LPS/TNF- significantly increased superoxide production, nuclear factor-
B activity, VCAM-1 expression, and endothelial adhesiveness for monocytes. Previous exposure to fluid flow inhibited these sequelae of exposure to cytokines or oxidized lipoprotein. The effect of fluid flow appears to be due in part to shear-induced release of NO, because coincubation with nitro-L-arginine completely abolished these effects of flow. Furthermore, the NO donor PAPA-NONOate and 8-Br-cGMP (but not 8-Br-cAMP) mimicked the effects of flow.
Conclusions Previous exposure to fluid flow decreased cytokine- or lipoprotein-stimulated endothelial cell superoxide production, VCAM-1 expression, and monocyte binding; the effects of flow appear to be due to NO. Flow-mediated NO-dependent regulation of oxidant-responsive transcription may influence the site of a lesion.
Key Words: blood flow • endothelium-derived factors • adhesion molecules • atherosclerosis • free radicals
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H. Matsushita, E. Chang, A. J. Glassford, J. P. Cooke, C.-P. Chiu, and P. S. Tsao eNOS Activity Is Reduced in Senescent Human Endothelial Cells: Preservation by hTERT Immortalization Circ. Res., October 26, 2001; 89(9): 793 - 798. [Abstract] [Full Text] [PDF]
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