Expression and Function of Recombinant Endothelial Nitric Oxide Synthase in Human Endothelial Cells

Abstract

Endothelial dysfunction is frequently involved in the pathogenesis of vascular disease. While nitric oxide (NO) inhibits smooth muscle cell proliferation, its effect on endothelial cell proliferation is unclear. The aim of this study was to determine if adenoviral-mediated gene transfer of endothelial NO synthase (eNOS) to human umbilical vein endothelial cells (HUVECs) would result in increased generation of NO and affect endothelial cell proliferation. HUVECs were transduced with adenoviral vectors encoding eNOS (AdeNOS) or β-galactosidase (Adβgal) or exposed to diluent (control). AdeNOS-transduced cells showed increased eNOS expression as detected by Western blot analysis, and increased concentrations of cGMP (control 0.7 ± 0.1; Adβgal 0.9 ± 0.2; AdeNOS 3.1 ± 0.5 pmol/mg protein; p < 0.001) and nitrite (control 11.8 ± 1.2; Adβgal 13.3 ± 1.7; AdeNOS 21.1 ± 2.2 nmol/mg protein/hour; p < 0.01). DNA synthesis as assessed by [³H]thymidine incorporation and cell counts were significantly reduced (by ∼30%) in AdeNOS-transduced HUVECs. Expression of mitogen-activated protein kinase was also decreased in AdeNOS-transduced cells. This study shows that adenoviral-mediated gene transfer of eNOS to HUVECs inhibits endothelial cell proliferation.

Journal

Journal ID (publisher-id): JVR

Journal ID (nlm-ta): J Vasc Res

Journal ID (doi): 10.1159/issn.1018-1172

Title: Journal of Vascular Research

Publisher: S. Karger AG

ISSN (Print): 1018-1172

ISSN (Electronic): 1423-0135

Publication date Subscription-year: 2000

Publication date (Print): December 2000

Publication date (Electronic): 10 January 2001

Volume: 37

Issue: 6

Pages: 449-456

Affiliations

Departments of ^aEndocrinology and ^bAnesthesiology, Mayo Clinic and Foundation, Rochester, Minn., USA

Article

Publisher ID: 54077 Other ID: J Vasc Res 2000;37:449–456

DOI: 10.1159/000054077

PubMed ID: 11146398

Copyright statement: © 2000 S. Karger AG, Basel

License:

Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher. Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug. Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.

Page count

Figures: 8, References: 27, Pages: 8

Product

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Categories

Subject: Research Paper