© 1999 American Heart Association, Inc.
Original Contribution |
Opposing Effects of Reactive Oxygen Species and Cholesterol on Endothelial Nitric Oxide Synthase and Endothelial Cell Caveolae
From the Department of Medicine (T.E.P., V.P., H.U., A.W.), Division of Cardiology, University of Washington, Seattle, Wash, and the Center for Cardiovascular Research (C.Y., B.C.B.), University of Rochester, Rochester, NY. The current affiliation for T.E.P. is the Department of Cardiovascular Diseases and the Molecular Medicine Program, Mayo Clinic, Rochester, Minn.
Correspondence to Bradford C. Berk, MD, PhD, University of Rochester Medical Center, Center for Cardiovascular Research, 601 Elmwood Ave, Box 679, Rochester, NY 14642. E-mail bradford_berk{at}urmc.rochester.edu
Abstract—Synthesis of nitric oxide (NO) by endothelial nitric oxide synthase (eNOS) is critical for normal vascular homeostasis. eNOS function is rapidly regulated by agonists and blood flow and chronically by factors that regulate mRNA stability and gene transcription. Recently, localization of eNOS to specialized plasma membrane invaginations termed caveolae has been proposed to be required for maximal eNOS activity. Because caveolae are highly enriched in cholesterol, and hypercholesterolemia is associated with increased NO production, we first studied the effects of cholesterol loading on eNOS localization and NO production in cultured bovine aortic endothelial cells (BAECs). Caveolae-enriched fractions were prepared by OptiPrep gradient density centrifugation. Treatment of BAECs with 30 µg/mL cholesterol for 24 hours stimulated significant increases in total eNOS protein expression (1.50-fold), eNOS associated with caveolae-enriched membranes (2.23-fold), and calcium ionophore-stimulated NO production (1.56-fold). Because reactive oxygen species (ROS) contribute to endothelial dysfunction in hypercholesterolemia, we next studied the effects of ROS on eNOS localization and caveolae number. Treatment of BAECs for 24 hours with 1 µmol/L LY83583, a superoxide-generating napthoquinolinedione, decreased caveolae number measured by electron microscopy and prevented the cholesterol-mediated increases in eNOS expression. In vitro exposure of caveolae-enriched membranes to ROS (xanthine plus xanthine oxidase) dissociated caveolin more readily than eNOS from the membranes. These results show that cholesterol treatment increases eNOS expression, whereas ROS treatment decreases eNOS expression and the association of eNOS with caveolin in caveolae-enriched membranes. Our data suggest that oxidative stress modulates endothelial function by regulating caveolae formation, eNOS expression, and eNOS-caveolin interactions.
Key Words: superoxide • caveolae • caveolin • eNOS • cholesterol
This article has been cited by other articles:
 |
|
 |
|
  R. Maas, E. Schwedhelm, L. Kahl, H. Li, R. Benndorf, N. Luneburg, U. Forstermann, and R. H. Boger Simultaneous Assessment of Endothelial Function, Nitric Oxide Synthase Activity, Nitric Oxide-Mediated Signaling, and Oxidative Stress in Individuals with and without Hypercholesterolemia Clin. Chem., February 1, 2008; 54(2): 292 - 300. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Kubo, T.-S. Li, R. Suzuki, B. Shirasawa, N. Morikage, M. Ohshima, S.-L. Qin, and K. Hamano Hypoxic preconditioning increases survival and angiogenic potency of peripheral blood mononuclear cells via oxidative stress resistance Am J Physiol Heart Circ Physiol, February 1, 2008; 294(2): H590 - H595. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. R. Duncan, S. J. Walker, V. A. Ezzat, S. B. Wheatcroft, J.-M. Li, A. M. Shah, and M. T. Kearney Accelerated endothelial dysfunction in mild prediabetic insulin resistance: the early role of reactive oxygen species Am J Physiol Endocrinol Metab, November 1, 2007; 293(5): E1311 - E1319. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 X. Wang, H. Mu, H. Chai, D. Liao, Q. Yao, and C. Chen Human Immunodeficiency Virus Protease Inhibitor Ritonavir Inhibits Cholesterol Efflux from Human Macrophage-Derived Foam Cells Am. J. Pathol., July 1, 2007; 171(1): 304 - 314. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Simionescu Implications of Early Structural-Functional Changes in the Endothelium for Vascular Disease Arterioscler. Thromb. Vasc. Biol., February 1, 2007; 27(2): 266 - 274. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Z.-G. Jin Where is endothelial nitric oxide synthase more critical: plasma membrane or Golgi? Arterioscler. Thromb. Vasc. Biol., May 1, 2006; 26(5): 959 - 961. [Full Text] [PDF]
|
|
|
|
 |
|
 K. HUSAIN, M. V. ORTIZ, and J. LALLA PHYSICAL TRAINING AMELIORATES CHRONIC ALCOHOL-INDUCED HYPERTENSION AND AORTIC REACTIVITY IN RATS Alcohol Alcohol., May 1, 2006; 41(3): 247 - 253. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Jacobi, S. Sela, H. I. Cohen, J. Chezar, and B. Kristal Priming of polymorphonuclear leukocytes: a culprit in the initiation of endothelial cell injury Am J Physiol Heart Circ Physiol, May 1, 2006; 290(5): H2051 - H2058. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 N. M. Gharavi, N. A. Baker, K. P. Mouillesseaux, W. Yeung, H. M. Honda, X. Hsieh, M. Yeh, E. J. Smart, and J. A. Berliner Role of Endothelial Nitric Oxide Synthase in the Regulation of SREBP Activation by Oxidized Phospholipids Circ. Res., March 31, 2006; 98(6): 768 - 776. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. M. Touyz Lipid Rafts Take Center Stage in Endothelial Cell Redox Signaling by Death Receptors Hypertension, January 1, 2006; 47(1): 16 - 18. [Full Text] [PDF]
|
|
|
|
 |
|
 H. Sies, W. Stahl, and A. Sevanian Nutritional, Dietary and Postprandial Oxidative Stress J. Nutr., May 1, 2005; 135(5): 969 - 972. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 X.-L. Wang, D. Ye, T. E. Peterson, S. Cao, V. H. Shah, Z. S. Katusic, G. C. Sieck, and H.-C. Lee Caveolae Targeting and Regulation of Large Conductance Ca2+-activated K+ Channels in Vascular Endothelial Cells J. Biol. Chem., March 25, 2005; 280(12): 11656 - 11664. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Arena, A. Pattarozzi, A. Corsaro, G. Schettini, and T. Florio Somatostatin Receptor Subtype-Dependent Regulation of Nitric Oxide Release: Involvement of Different Intracellular Pathways Mol. Endocrinol., January 1, 2005; 19(1): 255 - 267. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. O. Lungu, Z.-G. Jin, H. Yamawaki, T. Tanimoto, C. Wong, and B. C. Berk Cyclosporin A Inhibits Flow-mediated Activation of Endothelial Nitric-oxide Synthase by Altering Cholesterol Content in Caveolae J. Biol. Chem., November 19, 2004; 279(47): 48794 - 48800. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. He, T. E. Peterson, E. L. Holmuhamedov, A. Terzic, N. M. Caplice, L. W. Oberley, and Z. S. Katusic Human Endothelial Progenitor Cells Tolerate Oxidative Stress Due to Intrinsically High Expression of Manganese Superoxide Dismutase Arterioscler. Thromb. Vasc. Biol., November 1, 2004; 24(11): 2021 - 2027. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Sonveaux, P. Martinive, J. DeWever, Z. Batova, G. Daneau, M. Pelat, P. Ghisdal, V. Gregoire, C. Dessy, J.-L. Balligand, et al. Caveolin-1 Expression Is Critical for Vascular Endothelial Growth Factor-Induced Ischemic Hindlimb Collateralization and Nitric Oxide-Mediated Angiogenesis Circ. Res., July 23, 2004; 95(2): 154 - 161. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J.-P. Gratton, P. Bernatchez, and W. C. Sessa Caveolae and Caveolins in the Cardiovascular System Circ. Res., June 11, 2004; 94(11): 1408 - 1417. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. M. Reid and B. T. Kinsella The {alpha}, but Not the {beta}, Isoform of the Human Thromboxane A2 Receptor Is a Target for Nitric Oxide-mediated Desensitization: INDEPENDENT MODULATION OF TP{alpha} SIGNALING BY NITRIC OXIDE AND PROSTACYCLIN J. Biol. Chem., December 19, 2003; 278(51): 51190 - 51202. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Chatterjee, S. Cao, T. E. Peterson, R. D. Simari, and V. Shah Inhibition of GTP-dependent vesicle trafficking impairs internalization of plasmalemmal eNOS and cellular nitric oxide production J. Cell Sci., September 1, 2003; 116(17): 3645 - 3655. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
W. Tan, S. Rouen, K. M. Barkus, Y. S. Dremina, D. Hui, J. A. Christianson, D. E. Wright, S. O. Yoon, and R. T. Dobrowsky Nerve Growth Factor Blocks the Glucose-induced Down-regulation of Caveolin-1 Expression in Schwann Cells via p75 Neurotrophin Receptor Signaling J. Biol. Chem., June 13, 2003; 278(25): 23151 - 23162. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. Komers and S. Anderson Paradoxes of nitric oxide in the diabetic kidney Am J Physiol Renal Physiol, June 1, 2003; 284(6): F1121 - F1137. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. Gerzanich, S. Ivanova, H. Zhou, and J. M. Simard Mislocalization of eNOS and Upregulation of Cerebral Vascular Ca2+ Channel Activity in Angiotensin-Hypertension Hypertension, May 1, 2003; 41(5): 1124 - 1130. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Tanimoto, Z.-G. Jin, and B. C. Berk Transactivation of Vascular Endothelial Growth Factor (VEGF) Receptor Flk-1/KDR Is Involved in Sphingosine 1-Phosphate-stimulated Phosphorylation of Akt and Endothelial Nitric-oxide Synthase (eNOS) J. Biol. Chem., November 1, 2002; 277(45): 42997 - 43001. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Sun, J. Zhang, L. Lu, S. S. Chen, M. T. Quinn, and K. T. Weber Aldosterone-Induced Inflammation in the Rat Heart : Role of Oxidative Stress Am. J. Pathol., November 1, 2002; 161(5): 1773 - 1781. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y. Shizukuda and P. M. Buttrick Oxygen free radicals and heart failure: new insight into an old question Am J Physiol Lung Cell Mol Physiol, August 1, 2002; 283(2): L237 - L238. [Full Text] [PDF]
|
|
|
|
 |
|
 M. Annuk, M. Zilmer, L. Lind, T. Linde, and B. Fellstrom Oxidative Stress and Endothelial Function in Chronic Renal Failure J. Am. Soc. Nephrol., December 1, 2001; 12(12): 2747 - 2752. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. A. Jaimes, C. Sweeney, and L. Raij Effects of the Reactive Oxygen Species Hydrogen Peroxide and Hypochlorite on Endothelial Nitric Oxide Production Hypertension, October 1, 2001; 38(4): 877 - 883. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. E. von der Leyen and V. J. Dzau Therapeutic Potential of Nitric Oxide Synthase Gene Manipulation Circulation, June 5, 2001; 103(22): 2760 - 2765. [Full Text] [PDF]
|
|
|
|
|
|
R. Govers and T. J. Rabelink Cellular regulation of endothelial nitric oxide synthase Am J Physiol Renal Physiol, February 1, 2001; 280(2): F193 - F206. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Z.-G. Jin, M. G. Melaragno, D.-F. Liao, C. Yan, J. Haendeler, Y.-A. Suh, J. D. Lambeth, and B. C. Berk Cyclophilin A Is a Secreted Growth Factor Induced by Oxidative Stress Circ. Res., October 27, 2000; 87(9): 789 - 796. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. M. Channon, H. Qian, and S. E. George Nitric Oxide Synthase in Atherosclerosis and Vascular Injury : Insights From Experimental Gene Therapy Arterioscler. Thromb. Vasc. Biol., August 1, 2000; 20(8): 1873 - 1881. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. J. Guzik, N. E. J. West, E. Black, D. McDonald, C. Ratnatunga, R. Pillai, and K. M. Channon Vascular Superoxide Production by NAD(P)H Oxidase : Association With Endothelial Dysfunction and Clinical Risk Factors Circ. Res., May 12, 2000; 86 (9): e85 - e90. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J.-P. Gratton, J. Fontana, D. S. O'Connor, G. Garcia-Cardena, T. J. McCabe, and W. C. Sessa Reconstitution of an Endothelial Nitric-oxide Synthase (eNOS), hsp90, and Caveolin-1 Complex in Vitro. EVIDENCE THAT hsp90 FACILITATES CALMODULIN STIMULATED DISPLACEMENT OF eNOS FROM CAVEOLIN-1 J. Biol. Chem., July 14, 2000; 275(29): 22268 - 22272. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Cao, J. Yao, T. J. McCabe, Q. Yao, Z. S. Katusic, W. C. Sessa, and V. Shah Direct Interaction between Endothelial Nitric-oxide Synthase and Dynamin-2. IMPLICATIONS FOR NITRIC-OXIDE SYNTHASE FUNCTION J. Biol. Chem., April 20, 2001; 276(17): 14249 - 14256. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Nuszkowski, R. Grabner, G. Marsche, A. Unbehaun, E. Malle, and R. Heller Hypochlorite-modified Low Density Lipoprotein Inhibits Nitric Oxide Synthesis in Endothelial Cells via an Intracellular Dislocalization of Endothelial Nitric-oxide Synthase J. Biol. Chem., April 20, 2001; 276(17): 14212 - 14221. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Hoffmann, J. Haendeler, A. M. Zeiher, and S. Dimmeler TNFalpha and oxLDL Reduce Protein S-Nitrosylation in Endothelial Cells J. Biol. Chem., October 26, 2001; 276(44): 41383 - 41387. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. J. Guzik, S. Mussa, D. Gastaldi, J. Sadowski, C. Ratnatunga, R. Pillai, and K. M. Channon Mechanisms of Increased Vascular Superoxide Production in Human Diabetes Mellitus: Role of NAD(P)H Oxidase and Endothelial Nitric Oxide Synthase Circulation, April 9, 2002; 105(14): 1656 - 1662. [Abstract] [Full Text] [PDF]
|
|