EXPERIMENTAL STUDY
An endogenous inhibitor of nitric oxide synthase regulates endothelial adhesiveness for monocytes
Rainer H. Böger, MDa,
Stefanie M. Bode-Böger, MDa,
Philip S. Tsao, PhDa,
Patrick S. Lin, BSa,
Jason R. Chan, BSa and
John P. Cooke, MD, PhDa
a Section of Vascular Medicine, Stanford University School of Medicine, Stanford, California, USA
Manuscript received August 10, 1999;
revised manuscript received June 16, 2000,
accepted August 16, 2000.
Reprint requests and correspondence to: Dr. John P. Cooke, Division of Cardiovascular Medicine, Stanford University School of Medicine, 300 Pasteur Drive, Stanford, California 94305-5406
john.cooke{at}stanford.edu
OBJECTIVES
We sought to determine whether asymmetric dimethylarginine (ADMA) inhibits nitric oxide (NO) elaboration in cultured human endothelial cells and whether this is associated with the activation of oxidant-sensitive signaling mediating endothelial adhesiveness for monocytes.
BACKGROUND
Endothelial NO elaboration is impaired in hypercholesterolemia and atherosclerosis, which may be due to elevated concentrations of ADMA, an endogenous inhibitor of NO synthase.
METHODS
Human umbilical vein endothelial cells (ECV 304) and human monocytoid cells (THP-1) were studied in a functional binding assay. Nitric oxide and superoxide anion (O2–) were measured by chemiluminescence; ADMA by high pressure liquid chromatography; monocyte chemotactic protein-1 (MCP-1) by ELISA and NF- B by electromobility gel shift assay.
RESULTS
Incubation of endothelial cells with ADMA (0.1 µM to 100 µM) inhibited NO formation, which was reversed by coincubation with L-arginine (1 mM). The biologically inactive stereoisomer symmetric dimethylarginine did not inhibit NO release. Asymmetric dimethylarginine (10 µM) or native low-density lipoprotein cholesterol (100 mg/dL) increased endothelial O2– to the same degree. Asymmetric dimethylarginine also stimulated MCP-1 formation by endothelial cells. This effect was paralleled by activation of the redox-sensitive transcription factor NF-B. Preincubation of endothelial cells with ADMA increased the adhesiveness of endothelial cells for THP-1 cells in a concentration-dependent manner. Asymmetric dimethylarginine-induced monocyte binding was diminished by L-arginine or by a neutralizing anti-MCP-1 antibody.
CONCLUSIONS
We concluded that the endogenous NO synthase inhibitor ADMA is synthesized in human endothelial cells. Asymmetric dimethylarginine increases endothelial oxidative stress and potentiates monocyte binding. Asymmetric dimethylarginine may be an endogenous proatherogenic molecule.
|
Abbreviations and Acronyms
| | ADMA | = asymmetric dimethylarginine | | BHT | = butylated hydroxytoluene | | DDAH | = dimethylarginine dimethylaminohydrolase | | ECV 304 | = a line of transformed human umbilical vein endothelial cells | | ELISA | = enzyme-linked immunosorbent assay | | HBSS | = Hanks balanced salt solution | | HPLC | = high-pressure liquid chromatography | | LDL | = low-density lipoprotein | | LNMMA | = L, N-monomethylarginine | | MCP-1 | = monocyte chemotactic protein | | nLDL | = native low-density lipoprotein | | NO | = nitric oxide | | OPA | = o-phthaldialdehyde | | oxLDL | = oxidized low-density lipoprotein | | SDMA | = symmetric dimethylarginine | | THP-1 | = a line of human monocytoid cells |
|
This article has been cited by other articles:
|
|
|
|
 
E.-C. von Leitner, A. Klinke, D. Atzler, J. L. Slocum, N. Lund, J. T. Kielstein, R. Maas, R. Schmidt-Haupt, M. Pekarova, O. Hellwinkel, et al.
Pathogenic Cycle Between the Endogenous Nitric Oxide Synthase Inhibitor Asymmetrical Dimethylarginine and the Leukocyte-Derived Hemoprotein Myeloperoxidase
Circulation,
December 13, 2011;
124(24):
2735 - 2745.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. S. Pullamsetti, R. Savai, M. B. Schaefer, J. Wilhelm, H. A. Ghofrani, N. Weissmann, C. Schudt, I. Fleming, K. Mayer, J. Leiper, et al.
cAMP Phosphodiesterase Inhibitors Increases Nitric Oxide Production by Modulating Dimethylarginine Dimethylaminohydrolases
Circulation,
March 22, 2011;
123(11):
1194 - 1204.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
D. Zakrzewicz, A. Zakrzewicz, S. Wilker, R.-H. Boedeker, W. Padberg, O. Eickelberg, and V. Grau
Dimethylarginine metabolism during acute and chronic rejection of rat renal allografts
Nephrol. Dial. Transplant.,
January 1, 2011;
26(1):
124 - 135.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
E. Klein, J. Weigel, M. C. Buford, A. Holian, and S. M. Wells
Asymmetric dimethylarginine potentiates lung inflammation in a mouse model of allergic asthma
Am J Physiol Lung Cell Mol Physiol,
December 1, 2010;
299(6):
L816 - L825.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. G. Ayer, J. A. Harmer, S. Nakhla, W. Xuan, M. K.C. Ng, O. T. Raitakari, G. B. Marks, and D. S. Celermajer
HDL-Cholesterol, Blood Pressure, and Asymmetric Dimethylarginine Are Significantly Associated With Arterial Wall Thickness in Children
Arterioscler Thromb Vasc Biol,
June 1, 2009;
29(6):
943 - 949.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C. Antoniades, C. Shirodaria, P. Leeson, A. Antonopoulos, N. Warrick, T. Van-Assche, C. Cunnington, D. Tousoulis, R. Pillai, C. Ratnatunga, et al.
Association of plasma asymmetrical dimethylarginine (ADMA) with elevated vascular superoxide production and endothelial nitric oxide synthase uncoupling: implications for endothelial function in human atherosclerosis
Eur. Heart J.,
May 1, 2009;
30(9):
1142 - 1150.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
Q. G. Zhou, M. Zhou, F. F. Hou, and X. Peng
Asymmetrical dimethylarginine triggers lipolysis and inflammatory response via induction of endoplasmic reticulum stress in cultured adipocytes
Am J Physiol Endocrinol Metab,
April 1, 2009;
296(4):
E869 - E878.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. M. Wells, M. C. Buford, C. T. Migliaccio, and A. Holian
Elevated Asymmetric Dimethylarginine Alters Lung Function and Induces Collagen Deposition in Mice
Am. J. Respir. Cell Mol. Biol.,
February 1, 2009;
40(2):
179 - 188.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C.-C. Wu, S.-C. Wen, C.-W. Yang, S.-Y. Pu, K.-C. Tsai, and J.-W. Chen
Plasma ADMA Predicts Restenosis of Arteriovenous Fistula
J. Am. Soc. Nephrol.,
January 1, 2009;
20(1):
213 - 222.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
Z. Veresh, A. Racz, G. Lotz, and A. Koller
ADMA Impairs Nitric Oxide-Mediated Arteriolar Function Due to Increased Superoxide Production by Angiotensin II-NAD(P)H Oxidase Pathway
Hypertension,
November 1, 2008;
52(5):
960 - 966.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
N. Sud, S. M. Wells, S. Sharma, D. A. Wiseman, J. Wilham, and S. M. Black
Asymmetric dimethylarginine inhibits HSP90 activity in pulmonary arterial endothelial cells: role of mitochondrial dysfunction
Am J Physiol Cell Physiol,
June 1, 2008;
294(6):
C1407 - C1418.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. Desai, Y. Zhao, and J. S. Warren
Human recombinant erythropoietin augments serum asymmetric dimethylarginine concentrations but does not compromise nitric oxide generation in mice
Nephrol. Dial. Transplant.,
May 1, 2008;
23(5):
1513 - 1520.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. Zeller, C. Korandji, J.-C. Guilland, P. Sicard, C. Vergely, L. Lorgis, J.-C. Beer, L. Duvillard, A.-C. Lagrost, D. Moreau, et al.
Impact of Asymmetric Dimethylarginine on Mortality After Acute Myocardial Infarction
Arterioscler Thromb Vasc Biol,
May 1, 2008;
28(5):
954 - 960.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
F. Palm, M. L. Onozato, Z. Luo, and C. S. Wilcox
Dimethylarginine dimethylaminohydrolase (DDAH): expression, regulation, and function in the cardiovascular and renal systems
Am J Physiol Heart Circ Physiol,
December 1, 2007;
293(6):
H3227 - H3245.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
K. Hasegawa, S. Wakino, S. Tatematsu, K. Yoshioka, K. Homma, N. Sugano, M. Kimoto, K. Hayashi, and H. Itoh
Role of Asymmetric Dimethylarginine in Vascular Injury in Transgenic Mice Overexpressing Dimethylarginie Dimethylaminohydrolase 2
Circ. Res.,
July 20, 2007;
101(2):
e2 - e10.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. M. Wells and A. Holian
Asymmetric Dimethylarginine Induces Oxidative and Nitrosative Stress in Murine Lung Epithelial Cells
Am. J. Respir. Cell Mol. Biol.,
May 1, 2007;
36(5):
520 - 528.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
E. Monsalve, P. J. Oviedo, M. A. Garcia-Perez, J. J. Tarin, A. Cano, and C. Hermenegildo
Estradiol counteracts oxidized LDL-induced asymmetric dimethylarginine production by cultured human endothelial cells
Cardiovasc Res,
January 1, 2007;
73(1):
66 - 72.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. Tanaka, K. Sydow, F. Gunawan, J. Jacobi, P. S. Tsao, R. C. Robbins, and J. P. Cooke
Dimethylarginine Dimethylaminohydrolase Overexpression Suppresses Graft Coronary Artery Disease
Circulation,
September 13, 2005;
112(11):
1549 - 1556.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
P. Ravani, G. Tripepi, F. Malberti, S. Testa, F. Mallamaci, and C. Zoccali
Asymmetrical Dimethylarginine Predicts Progression to Dialysis and Death in Patients with Chronic Kidney Disease: A Competing Risks Modeling Approach
J. Am. Soc. Nephrol.,
August 1, 2005;
16(8):
2449 - 2455.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. M Bode-Boger, F. Scalera, and J. Martens-Lobenhoffer
Asymmetric dimethylarginine (ADMA) accelerates cell senescence
Vascular Medicine,
July 1, 2005;
10(1_suppl):
S65 - S71.
[Abstract]
[PDF]
|
|
|
|
|
|
|
T. Teerlink
ADMA metabolism and clearance
Vascular Medicine,
July 1, 2005;
10(1_suppl):
S73 - S81.
[Abstract]
[PDF]
|
|
|
|
|
|
|
R. H Boger, E. Schwedhelm, R. Maas, S. Quispe-Bravo, and C. Skamira
ADMA and oxidative stress may relate to the progression of renal disease: rationale and design of the VIVALDI study
Vascular Medicine,
July 1, 2005;
10(1_suppl):
S97 - S102.
[Abstract]
[PDF]
|
|
|
|
|
|
|
S. M Bode-Boger, F. Scalera, and J. Martens-Lobenhoffer
Asymmetric dimethylarginine (ADMA) accelerates cell senescence
Vascular Medicine,
May 1, 2005;
10(2_suppl):
S65 - S71.
[Abstract]
[PDF]
|
|
|
|
|
|
|
T. Teerlink
ADMA metabolism and clearance
Vascular Medicine,
May 1, 2005;
10(2_suppl):
S73 - S81.
[Abstract]
[PDF]
|
|
|
|
|
|
|
R. H Boger, E. Schwedhelm, R. Maas, S. Quispe-Bravo, and C. Skamira
ADMA and oxidative stress may relate to the progression of renal disease: rationale and design of the VIVALDI study
Vascular Medicine,
May 1, 2005;
10(2_suppl):
S97 - S102.
[Abstract]
[PDF]
|
|
|
|
|
|
|
F. Scalera, J. Borlak, B. Beckmann, J. Martens-Lobenhoffer, T. Thum, M. Tager, and S. M. Bode-Boger
Endogenous Nitric Oxide Synthesis Inhibitor Asymmetric Dimethyl L-Arginine Accelerates Endothelial Cell Senescence
Arterioscler Thromb Vasc Biol,
October 1, 2004;
24(10):
1816 - 1822.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
I. V. Smirnova, M. Kajstura, T. Sawamura, and M. S. Goligorsky
Asymmetric dimethylarginine upregulates LOX-1 in activated macrophages: role in foam cell formation
Am J Physiol Heart Circ Physiol,
August 1, 2004;
287(2):
H782 - H790.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
P. Vallance and J. Leiper
Cardiovascular Biology of the Asymmetric Dimethylarginine:Dimethylarginine Dimethylaminohydrolase Pathway
Arterioscler Thromb Vasc Biol,
June 1, 2004;
24(6):
1023 - 1030.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. P. Cooke
Asymmetrical Dimethylarginine: The Uber Marker?
Circulation,
April 20, 2004;
109(15):
1813 - 1818.
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. Weis, T. N. Kledal, K. Y. Lin, S. N. Panchal, S. Z. Gao, H. A. Valantine, E. S. Mocarski, and J. P. Cooke
Cytomegalovirus Infection Impairs the Nitric Oxide Synthase Pathway: Role of Asymmetric Dimethylarginine in Transplant Arteriosclerosis
Circulation,
February 3, 2004;
109(4):
500 - 505.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
H. Dayoub, V. Achan, S. Adimoolam, J. Jacobi, M. C. Stuehlinger, B.-y. Wang, P. S. Tsao, M. Kimoto, P. Vallance, A. J. Patterson, et al.
Dimethylarginine Dimethylaminohydrolase Regulates Nitric Oxide Synthesis: Genetic and Physiological Evidence
Circulation,
December 16, 2003;
108(24):
3042 - 3047.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
T.-M. Lu, Y.-A. Ding, S.-J. Lin, W.-S. Lee, and H.-C. Tai
Plasma levels of asymmetrical dimethylarginine and adverse cardiovascular events after percutaneous coronary intervention
Eur. Heart J.,
November 1, 2003;
24(21):
1912 - 1919.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
R. H Boger
The emerging role of asymmetric dimethylarginine as a novel cardiovascular risk factor
Cardiovasc Res,
October 1, 2003;
59(4):
824 - 833.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
R. H Boger
Asymmetric dimethylarginine (ADMA) modulates endothelial function - therapeutic implications
Vascular Medicine,
August 1, 2003;
8(3):
149 - 151.
[PDF]
|
|
|
|
|
|
|
M. Weis and J. P. Cooke
Cardiac Allograft Vasculopathy and Dysregulation of the NO Synthase Pathway
Arterioscler Thromb Vasc Biol,
April 1, 2003;
23(4):
567 - 575.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. R. Clark, M. J. Coffey, R. M. Maclean, P. W. Collins, M. J. Lewis, A. R. Cross, and V. B. O'Donnell
Characterization of Nitric Oxide Consumption Pathways by Normal, Chronic Granulomatous Disease and Myeloperoxidase-Deficient Human Neutrophils
J. Immunol.,
November 15, 2002;
169(10):
5889 - 5896.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
K. Y. Lin, A. Ito, T. Asagami, P. S. Tsao, S. Adimoolam, M. Kimoto, H. Tsuji, G. M. Reaven, and J. P. Cooke
Impaired Nitric Oxide Synthase Pathway in Diabetes Mellitus: Role of Asymmetric Dimethylarginine and Dimethylarginine Dimethylaminohydrolase
Circulation,
August 20, 2002;
106(8):
987 - 992.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
D. Tousoulis, C. Antoniades, C. Tentolouris, G. Goumas, C. Stefanadis, and P. Toutouzas
L-Arginine in cardiovascular disease: dream or reality?
Vascular Medicine,
August 1, 2002;
7(3):
203 - 211.
[Abstract]
[PDF]
|
|
|
|
|
|
|
D. T. Nash
Insulin Resistance, ADMA Levels, and Cardiovascular Disease
JAMA,
March 20, 2002;
287(11):
1451 - 1452.
[Full Text]
[PDF]
|
|
|
|
|
|
|
C. Zoccali, F. A. Benedetto, R. Maas, F. Mallamaci, G. Tripepi, L. Salvatore Malatino, and R. Boger
Asymmetric Dimethylarginine, C-Reactive Protein, and Carotid Intima-Media Thickness in End-Stage Renal Disease
J. Am. Soc. Nephrol.,
February 1, 2002;
13(2):
490 - 496.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. C. Stuhlinger, P. S. Tsao, J.-H. Her, M. Kimoto, R. F. Balint, and J. P. Cooke
Homocysteine Impairs the Nitric Oxide Synthase Pathway: Role of Asymmetric Dimethylarginine
Circulation,
November 20, 2001;
104(21):
2569 - 2575.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
