This Article Full Text Full Text (PDF) All Versions of this Article: 93/6/523    most recent 01.RES.0000091336.55487.F7v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Niwano, K. Articles by Kurabayashi, M. Search for Related Content PubMed PubMed Citation Articles by Niwano, K. Articles by Kurabayashi, M. Related Collections Cell signalling/signal transduction Gene expression Gene regulation Peripheral vascular disease Endothelium/vascular type/nitric oxide

(Circulation Research. 2003;93:523.)
© 2003 American Heart Association, Inc.

Molecular Medicine

Transcriptional Stimulation of the eNOS Gene by the Stable Prostacyclin Analogue Beraprost Is Mediated Through cAMP-Responsive Element in Vascular Endothelial Cells

Close Link Between PGI₂ Signal and NO Pathways

Kazuo Niwano, Masashi Arai, Koichi Tomaru, Tsuyoshi Uchiyama, Yoshio Ohyama, Masahiko Kurabayashi

From the Second Department of Internal Medicine, Gunma University School of Medicine, Japan.

Correspondence to Masahiko Kurabayashi, MD, PhD, Second Department of Internal Medicine, Gunma University School of Medicine, 3-39-15 Showa-machi, Maebashi, Gunma 371-8511, Japan. E-mail mkuraba{at}med.gunma-u.ac.jp

Beraprost sodium (BPS), an orally active prostacyclin analogue, has been reported to be beneficial in the treatment of primary pulmonary hypertension and obstructive peripheral arterial disease. Although BPS was originally described for its effects on platelet aggregation and vasodilatory response, the effect on endothelial cells has been poorly understood. In this study, we examined the effects of BPS on the eNOS gene expression in mouse aorta and cultured human and bovine aortic endothelial cells. Treatment of these cells with BPS increased the eNOS expression as assessed by Northern blots, Western blots, and NO production by NO-specific fluorescence (DAF2-DA) and by the Griess method. Standard mRNA decay assays showed that BPS increases the stability of eNOS mRNA. In addition, BPS increased the promoter activity of the human eNOS gene, as determined by luciferase assays of the eNOS promoter gene. Progressive 5'-deletion and site-specific mutation analyses defined the BPS-responsive sequences as cAMP-responsive elements (CRE) located at -733 and -603. By using the oligonucleotide probe containing this CRE sequence in electrophoretic mobility shift assays, we showed that the phosphorylated form of CRE-binding protein is a major constituent of the complex in BPS-treated cells. Western blot analyses indicate that BPS but not endogenous prostacyclin phosphorylates CRE-binding protein. The presence of functional CRE sites within human eNOS promoter may represent a novel mechanism for regulating eNOS gene expression.

Key Words: endothelium • prostaglandins • nitric oxide • nitric oxide synthase

This article has been cited by other articles:

				Y.-M. Sue, C.-P. Chung, H. Lin, Y. Chou, C.-Y. Jen, H.-F. Li, C.-C. Chang, and S.-H. Juan PPAR{delta}-mediated p21/p27 induction via increased CREB-binding protein nuclear translocation in beraprost-induced antiproliferation of murine aortic smooth muscle cells Am J Physiol Cell Physiol, August 1, 2009; 297(2): C321 - C329. [Abstract] [Full Text] [PDF]

				A. Barac and J. A. Panza Mechanisms of Decreased Vascular Function With Aging Hypertension, June 1, 2009; 53(6): 900 - 902. [Full Text] [PDF]

				H. Si and D. Liu Genistein, a Soy Phytoestrogen, Upregulates the Expression of Human Endothelial Nitric Oxide Synthase and Lowers Blood Pressure in Spontaneously Hypertensive Rats J. Nutr., February 1, 2008; 138(2): 297 - 304. [Abstract] [Full Text] [PDF]

				N. Toda, K. Ayajiki, and T. Okamura Interaction of Endothelial Nitric Oxide and Angiotensin in the Circulation Pharmacol. Rev., March 1, 2007; 59(1): 54 - 87. [Abstract] [Full Text] [PDF]

				E. M. Repetto, V. Pannunzio, F. Astort, C. Martinez Calejman, M. B. Moreno, O. P. Pignataro, and C. B. Cymeryng Characterization of L-arginine transport in adrenal cells: effect of ACTH Am J Physiol Endocrinol Metab, August 1, 2006; 291(2): E291 - E297. [Abstract] [Full Text] [PDF]

				K. Niwano, M. Arai, N. Koitabashi, S. Hara, A. Watanabe, K. Sekiguchi, T. Tanaka, T. Iso, and M. Kurabayashi Competitive Binding of CREB and ATF2 to cAMP/ATF Responsive Element Regulates eNOS Gene Expression in Endothelial Cells Arterioscler Thromb Vasc Biol, May 1, 2006; 26(5): 1036 - 1042. [Abstract] [Full Text] [PDF]

				E. M. Antman, D. DeMets, and J. Loscalzo Cyclooxygenase Inhibition and Cardiovascular Risk Circulation, August 2, 2005; 112(5): 759 - 770. [Full Text] [PDF]

				Y. Lamarche, O. Malo, E. Thorin, A. Denault, M. Carrier, J. Roy, and L.P. Perrault Inhaled but not intravenous milrinone prevents pulmonary endothelial dysfunction after cardiopulmonary bypass J. Thorac. Cardiovasc. Surg., July 1, 2005; 130(1): 83 - 92. [Abstract] [Full Text] [PDF]

				N. Kawada, G. Solis, N. Ivey, S. Connors, K. Dennehy, P. Modlinger, R. Hamel, J. T. Kawada, E. Imai, R. Langenbach, et al. Cyclooxygenase-1-Deficient Mice Have High Sleep-to-Wake Blood Pressure Ratios and Renal Vasoconstriction Hypertension, June 1, 2005; 45(6): 1131 - 1138. [Abstract] [Full Text] [PDF]

				D. Liu, H. Jiang, and R. W. Grange Genistein Activates the 3',5'-Cyclic Adenosine Monophosphate Signaling Pathway in Vascular Endothelial Cells and Protects Endothelial Barrier Function Endocrinology, March 1, 2005; 146(3): 1312 - 1320. [Abstract] [Full Text] [PDF]