From the Department of Pharmacology, Johannes Gutenberg University,
Mainz, Germany.
Correspondence and reprint requests to Dr Ulrich Förstermann, Department of Pharmacology, Johannes Gutenberg University, Obere Zahlbacher Strasse 67, D-55101 Mainz, Germany. E-mail ulrich.forstermann{at}uni-mainz.de
Abstract—Estrogens have been found
to reduce the incidence of cardiovascular disease that
has been ascribed in part to an increased expression and/or activity of
the vasoprotective endothelial NO synthase (NOS III).
Some reports have shown that the level of expression of this
constitutive enzyme can be upregulated by estrogens. The current study
investigates the molecular mechanism of the NOS III upregulation in
human endothelial EA.hy 926 cells. Incubation of EA.hy
926 cells with 17ß-estradiol or the more stable 17
© 1998 American Heart Association, Inc.
Scientific Contributions
Estrogens Increase Transcription of the Human Endothelial NO Synthase Gene
Analysis of the Transcription Factors Involved
-ethinyl
estradiol enhanced NOS III mRNA and protein expression up to 1.8-fold,
without changing the stability of the NOS III mRNA. There was no
enhancement of NOS III mRNA after incubation of EA.hy 926 cells with
testosterone, progesterone, or dihydrocortisol or when 17-ethinyl
estradiol was added together with the estrogen antagonist
RU58668, indicating a specific estrogenic response. Nuclear run-on
assays indicated that the increase in NOS III mRNA is the result of an
estrogen-induced enhancement of NOS III gene transcription. In
transient transfection experiments using a 1.6 kb human NOS III
promoter fragment (which contains no bona fide estrogen-responsive
element, ERE), basal promoter activity was enhanced 1.7-fold by
17-ethinyl estradiol. In electrophoretic mobility shift assays,
nuclear extracts from estrogen-incubated EA.hy 926 cells showed no
enhanced binding activity either for the ERE-like motif in the human
NOS III promoter or for transcription factor GATA. However, binding of
transcription factor Sp1 (which is essential for the activity of the
human NOS III promoter) was significantly enhanced by estrogens. These
data suggest that the estrogen stimulation of the NOS III promoter
could be mediated in part by an increased activity of transcription
factor Sp1.
Key Words: 17-ethinyl estradiol • 17ß-estradiol • nitric oxide synthase • transcription factor Sp1
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H. J. Knot, K. M. Lounsbury, J. E. Brayden, and M. T. Nelson Gender differences in coronary artery diameter reflect changes in both endothelial Ca2+ and ecNOS activity Am J Physiol Heart Circ Physiol, March 1, 1999; 276(3): H961 - H969. [Abstract] [Full Text] [PDF]
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M. L Kuroski de Bold Estrogen, natriuretic peptides and the renin-angiotensin system Cardiovasc Res, March 1, 1999; 41(3): 524 - 531. [Abstract] [Full Text] [PDF]
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M.A Barbacanne, J Rami, J.B Michel, J.P Souchard, M Philippe, J.P Besombes, F Bayard, and J.F Arnal Estradiol increases rat aorta endothelium-derived relaxing factor (EDRF) activity without changes in endothelial NO synthase gene expression: possible role of decreased endothelium-derived superoxide anion production Cardiovasc Res, March 1, 1999; 41(3): 672 - 681. [Abstract] [Full Text] [PDF]
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P. M. Schwarz, B. Gierten, J.-P. Boissel, and U. Förstermann Expressional Down-Regulation of Neuronal-Type Nitric Oxide Synthase I by Glucocorticoids in N1E-115 Neuroblastoma Cells Mol. Pharmacol., August 1, 1998; 54(2): 258 - 263. [Abstract] [Full Text]
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X. Wang and A. A. Abdel-Rahman Estrogen modulation of eNOS activity and its association with caveolin-3 and calmodulin in rat hearts Am J Physiol Heart Circ Physiol, June 1, 2002; 282(6): H2309 - H2315. [Abstract] [Full Text] [PDF]
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Y. Chu, D. D. Heistad, K. L. Knudtson, K. G. Lamping, and F. M. Faraci Quantification of mRNA for Endothelial NO Synthase in Mouse Blood Vessels by Real-Time Polymerase Chain Reaction Arterioscler Thromb Vasc Biol, April 1, 2002; 22(4): 611 - 616. [Abstract] [Full Text] [PDF]
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F. Gobeil Jr, I. Dumont, A. M. Marrache, A. Vazquez-Tello, S. G. Bernier, D. Abran, X. Hou, M. H. Beauchamp, C. Quiniou, A. Bouayad, et al. Regulation of eNOS Expression in Brain Endothelial Cells by Perinuclear EP3 Receptors Circ. Res., April 5, 2002; 90(6): 682 - 689. [Abstract] [Full Text] [PDF]
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