© 1996 American Heart Association, Inc.
Articles |
Reduced Gene Expression of Vascular Endothelial NO Synthase and Cyclooxygenase-1 in Heart Failure
From the Departments of Experimental Pathology (C.J.S., B.S.R., Y.K., K.P.) and Physiology (D.S., C.H., X.Z., G.Z., X.-B.X., T.H.H.), New York Medical College, Valhalla, NY, and the Department of Molecular Cardiobiology (W.C.S.), Boyer Center for Molecular Medicine, Yale University School of Medicine, New Haven, Conn.
Correspondence to Thomas H. Hintze, PhD, Professor, Department of Physiology, New York Medical College, Valhalla, NY 10595.
Abstract Endothelium-dependent responses are depressed in coronary and peripheral blood vessels after the onset of pacing-induced heart failure in dogs and heart failure of various etiologies in humans. The present study was designed to examine whether these responses were due to decreases in the expression of endothelial cell NO synthase (ecNOS) and cyclooxygenase-1 (COX-1). After 1 month of left ventricular pacing, 8 mongrel dogs were monitored for heart failure as defined by clinical signs and left ventricular end diastolic pressures >25 mm Hg. Total RNA and protein were isolated from endothelial cells scraped from the thoracic aorta and analyzed by Northern and Western blotting, respectively. Blots probed with 32P-labeled cDNAs for ecNOS and COX-1 were quantified densitometrically, and results were normalized against GAPDH or von Willebrand factor (vWF). In arbitrary units, the ratios of ecNOS to GAPDH were 2.66±0.77 (mean±SEM, n=17) and 1.12±0.37 (n=6), and the ratios of COX-1 to GAPDH were 1.52±0.52 and 0.56±0.15 before and after heart failure, respectively. These represent 56% to 64% (P<.05) reductions in ecNOS and COX-1 gene expression. There was no change in the ratios of either COX-1 or ecNOS to vWF. There was also a marked reduction in ecNOS protein after heart failure, estimated at 70%. A marked reduction in nitrite production, a measure of enzyme activity, from thoracic aortas in response to stimulation by either acetylcholine or bradykinin also occurred. To determine whether ecNOS and COX-1 could be independently regulated, an orally active NO-releasing agent, CAS 936, was given to 7 normal dogs for 7 days, and aortic ecNOS and COX-1 mRNAs were analyzed. The ratio of ecNOS to GAPDH was depressed by 52% (P<.05) in aortas from these dogs, whereas the ratio of COX-1 to GAPDH was unchanged. Similar results were found when data were normalized to vWF. These results suggest that at least two endothelial vasodilator gene products are reduced in heart failure, as opposed to a selective defect in NO synthase gene expression.
Key Words: GAPDH • endothelial cell NO synthase • cyclooxygenase-1 • von Willebrand factor • Northern blots
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B. Hornig, N. Arakawa, D. Haussmann, and H. Drexler Differential Effects of Quinaprilat and Enalaprilat on Endothelial Function of Conduit Arteries in Patients With Chronic Heart Failure Circulation, December 22, 1998; 98(25): 2842 - 2848. [Abstract] [Full Text]
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P. Magri, M. A. E. Rao, S. Cangianiello, V. Bellizzi, R. Russo, A. F. Mele, M. Andreucci, B. Memoli, Luca De Nicola, and M. Volpe Early Impairment of Renal Hemodynamic Reserve in Patients With Asymptomatic Heart Failure Is Restored by Angiotensin II Antagonism Circulation, December 22, 1998; 98(25): 2849 - 2854. [Abstract] [Full Text] [PDF]
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H. Drexler Endothelium as a Therapeutic Target in Heart Failure Circulation, December 15, 1998; 98(24): 2652 - 2655. [Full Text] [PDF]
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F. H. Khadour, D. W. O'Brien, Y. Fu, P. W. Armstrong, and R. Schulz Endothelial nitric oxide synthase increases in left atria of dogs with pacing-induced heart failure Am J Physiol Heart Circ Physiol, December 1, 1998; 275(6): H1971 - H1978. [Abstract] [Full Text] [PDF]
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F. A. Recchia, P. I. McConnell, R. D. Bernstein, T. R. Vogel, X. Xu, and T. H. Hintze Reduced Nitric Oxide Production and Altered Myocardial Metabolism During the Decompensation of Pacing-Induced Heart Failure in the Conscious Dog Circ. Res., November 16, 1998; 83(10): 969 - 979. [Abstract] [Full Text] [PDF]
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B. Stein, T. Eschenhagen, J. Rudiger, H. Scholz, U. Forstermann, and I. Gath Increased expression of constitutive nitric oxide synthase III, but not inducible nitric oxide synthase II, in human heart failure J. Am. Coll. Cardiol., November 1, 1998; 32(5): 1179 - 1186. [Abstract] [Full Text] [PDF]
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W. Wang Cardiac sympathetic afferent stimulation by bradykinin in heart failure: role of NO and prostaglandins Am J Physiol Heart Circ Physiol, September 1, 1998; 275(3): H783 - H788. [Abstract] [Full Text] [PDF]
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S M Maguire, A G Nugent, C McGurk, G D Johnston, and D P Nicholls Abnormal vascular responses in human chronic cardiac failure are both endothelium dependent and endothelium independent Heart, August 1, 1998; 80(2): 141 - 145. [Abstract] [Full Text]
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Q. Feng, X. Lu, A. J Fortin, A. Pettersson, T. Hedner, R. L Kline, and J.M. O Arnold Elevation of an endogenous inhibitor of nitric oxide synthesis in experimental congestive heart failure Cardiovasc Res, March 1, 1998; 37(3): 667 - 675. [Abstract] [Full Text] [PDF]
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X. Zhang and T. H. Hintze Amlodipine Releases Nitric Oxide From Canine Coronary Microvessels : An Unexpected Mechanism of Action of a Calcium Channel–Blocking Agent Circulation, February 17, 1998; 97(6): 576 - 580. [Abstract] [Full Text] [PDF]
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B. Hornig, N. Arakawa, C. Kohler, and H. Drexler Vitamin C Improves Endothelial Function of Conduit Arteries in Patients With Chronic Heart Failure Circulation, February 3, 1998; 97(4): 363 - 368. [Abstract] [Full Text] [PDF]
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F. M. FARACI and D. D. HEISTAD Regulation of the Cerebral Circulation: Role of Endothelium and Potassium Channels Physiol Rev, January 1, 1998; 78(1): 53 - 97. [Abstract] [Full Text] [PDF]
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R. H. Ritchie, J. D. Marsh, W. D. Lancaster, C. A. Diglio, and R. J. Schiebinger Bradykinin Blocks Angiotensin II-Induced Hypertrophy in the Presence of Endothelial Cells Hypertension, January 1, 1998; 31(1): 39 - 44. [Abstract] [Full Text]
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K. Bhagat and P. Vallance Inflammatory Cytokines Impair Endothelium-Dependent Dilatation in Human Veins In Vivo Circulation, November 4, 1997; 96(9): 3042 - 3047. [Abstract] [Full Text]
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C. J. Smith, R. Huang, D. Sun, S. Ricketts, C. Hoegler, J.-Z. Ding, R. A. Moggio, and T. H. Hintze Development of Decompensated Dilated Cardiomyopathy Is Associated With Decreased Gene Expression and Activity of the Milrinone-Sensitive cAMP Phosphodiesterase PDE3A Circulation, November 4, 1997; 96(9): 3116 - 3123. [Abstract] [Full Text]
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B. D. Prendergast, B M. Sci, V. F. Sagach, and A. M. Shah Basal Release of Nitric Oxide Augments the Frank-Starling Response in the Isolated Heart Circulation, August 19, 1997; 96(4): 1320 - 1329. [Abstract] [Full Text]
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G. Zhao, X. Zhang, X. Xu, M. Ochoa, and T. H. Hintze Short-term Exercise Training Enhances Reflex Cholinergic Nitric Oxide–Dependent Coronary Vasodilation in Conscious Dogs Circ. Res., June 19, 1997; 80(6): 868 - 876. [Abstract] [Full Text]
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Y.-W. Xie, W. Shen, G. Zhao, X. Xu, M. S. Wolin, and T. H. Hintze Role of Endothelium-Derived Nitric Oxide in the Modulation of Canine Myocardial Mitochondrial Respiration In Vitro: Implications for the Development of Heart Failure Circ. Res., September 1, 1996; 79(3): 381 - 387. [Abstract] [Full Text]
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M. R. Kichuk, N. Seyedi, X. Zhang, C. C. Marboe, R. E. Michler, L. J. Addonizio, G. Kaley, A. Nasjletti, and T. H. Hintze Regulation of Nitric Oxide Production in Human Coronary Microvessels and the Contribution of Local Kinin Formation Circulation, July 1, 1996; 94(1): 44 - 51. [Abstract] [Full Text]
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D. B. Haitsma, D. Merkus, J. Vermeulen, P. D. Verdouw, and D. J. Duncker Nitric oxide production is maintained in exercising swine with chronic left ventricular dysfunction Am J Physiol Heart Circ Physiol, June 1, 2002; 282(6): H2198 - H2209. [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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