Amino Acids

, Volume 47, Issue 9, pp 1975–1982 | Cite as

Plasma asymmetric and symmetric dimethylarginine in a rat model of endothelial dysfunction induced by acute hyperhomocysteinemia

  • Joëlle Magné
  • Jean-François Huneau
  • Didier Borderie
  • Véronique Mathé
  • Cécile Bos
  • François Mariotti
Original Article
Part of the following topical collections:
  1. Homoarginine, Arginine and Relatives


Hyperhomocysteinemia induces vascular endothelial dysfunction, an early hallmark of atherogenesis. While higher levels of circulating asymmetric dimethylarginine (ADMA) and symmetric dimethyl arginine (SDMA), endogenous inhibitors of nitric oxide synthesis, have been associated with increased cardiovascular risk, the role that ADMA and SDMA play in the initiation of hyperhomocysteinemia-induced endothelial dysfunction remains still controversial. In the present study, we studied the changes of circulating ADMA and SDMA in a rat model of acutely hyperhomocysteinemia-induced endothelial dysfunction. In healthy rats, endothelium-related vascular reactivity (measured as acetylcholine-induced transient decrease in mean arterial blood pressure), plasma ADMA and SDMA, total plasma homocysteine (tHcy), cysteine and glutathione were measured before and 2, 4 and 6 h after methionine loading or vehicle. mRNA expression of hepatic dimethylarginine dimethylaminohydrolase-1 (DDAH1), a key protein responsible for ADMA metabolism, was measured 6 h after the methionine loading or the vehicle. Expectedly, methionine load induced a sustained increase in tHcy (up to 54.9 ± 1.9 µM) and a 30 % decrease in vascular reactivity compared to the baseline values. Plasma ADMA and SDMA decreased transiently after the methionine load. Hepatic mRNA expression of DDAH1, cathepsin D, and ubiquitin were significantly lower 6 h after the methionine load than after the vehicle. The absence of an elevation of circulating ADMA and SDMA in this model suggests that endothelial dysfunction induced by acute hyperhomocysteinemia cannot be explained by an up-regulation of protein arginine methyltransferases or a down-regulation of DDAH1. In experimental endothelial dysfunction induced by acute hyperhomocysteinemia, down-regulation of the proteasome is likely to dampen the release of ADMA and SDMA in the circulation.


ADMA Endothelial dysfunction Hyperhomocysteinemia Methionine load Proteolytic degradation SDMA 





Asymmetric dimethylarginine


Dimethylarginine dimethylaminohydrolase




Mean arterial pressure


Nitric oxide


Protein arginine methyltransferase


Reactive oxygen species




Symmetric dimethylarginine


Total plasma homocysteine



We thank Maëlle Robert for her contribution to the experimentation and Dominique Hermier for the help with the collection of the tissue samples. This work was supported by the French Ministry of Research. J. Magné is supported by the Swedish Heart–Lung Foundation, the Fredrik and Ingrid Thuring Foundation, and the Lars Hiertas Minne Foundation.

Conflict of interest

None of the authors had a conflict of interest.

Ethical approval

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving animals were in accordance with the ethical standards of the guidelines issued by the French National Animal Care Committee at which the studies were conducted.


  1. Andreotti F, Burzotta F, Manzoli A, Robinson K (2000) Homocysteine and risk of cardiovascular disease. J Thromb Thrombolysis 9(1):13–21CrossRefPubMedGoogle Scholar
  2. Antoniades C, Tousoulis D, Marinou K, Vasiliadou C, Tentolouris C, Bouras G, Pitsavos C, Stefanadis C (2006) Asymmetrical dimethylarginine regulates endothelial function in methionine-induced but not in chronic homocystinemia in humans: effect of oxidative stress and proinflammatory cytokines. Am J Clin Nutr 84(4):781–788PubMedGoogle Scholar
  3. Bellamy MF, McDowell IF, Ramsey MW, Brownlee M, Bones C, Newcombe RG, Lewis MJ (1998) Hyperhomocysteinemia after an oral methionine load acutely impairs endothelial function in healthy adults. Circulation 98(18):1848–1852CrossRefPubMedGoogle Scholar
  4. Bode-Böger SM, Scalera F, Kielstein JT, Martens-Lobenhoffer J, Breithardt G, Fobker M, Reinecke H (2006) Symmetrical dimethylarginine: a new combined parameter for renal function and extent of coronary artery disease. J Am Soc Nephrol 17(4):1128–1134. doi:10.1681/asn.2005101119 CrossRefPubMedGoogle Scholar
  5. Böger RH (2006) Asymmetric dimethylarginine (ADMA): a novel risk marker in cardiovascular medicine and beyond. Ann Med 38(2):126–136. doi:10.1080/07853890500472151 CrossRefPubMedGoogle Scholar
  6. Böger RH, Bode-Böger SM, Sydow K, Heistad DD, Lentz SR (2000) Plasma concentration of asymmetric dimethylarginine, an endogenous inhibitor of nitric oxide synthase, is elevated in monkeys with hyperhomocyst(e)inemia or hypercholesterolemia. Arterioscler Thromb Vasc Biol 20(6):1557–1564CrossRefPubMedGoogle Scholar
  7. Böger RH, Lentz SR, Bode-Böger SM, Knapp HR, Haynes WG (2001) Elevation of asymmetrical dimethylarginine may mediate endothelial dysfunction during experimental hyperhomocyst(e)inaemia in humans. Clin Sci (Lond) 100(2):161–167CrossRefGoogle Scholar
  8. Brosnan JT, da Silva R, Brosnan ME (2007) Amino acids and the regulation of methyl balance in humans. Curr Opin Clin Nutr Metab Care 10(1):52–57. doi:10.1097/MCO.0b013e3280110171 CrossRefPubMedGoogle Scholar
  9. Casas JP, Bautista LE, Smeeth L, Sharma P, Hingorani AD (2005) Homocysteine and stroke: evidence on a causal link from mendelian randomisation. Lancet 365(9455):224–232. doi:10.1016/s0140-6736(05)17742-3 CrossRefPubMedGoogle Scholar
  10. Chambers JC, Obeid OA, Kooner JS (1999) Physiological increments in plasma homocysteine induce vascular endothelial dysfunction in normal human subjects. Arterioscler Thromb Vasc Biol 19(12):2922–2927CrossRefPubMedGoogle Scholar
  11. Dayal S, Lentz SR (2008) Murine models of hyperhomocysteinemia and their vascular phenotypes. Arterioscler Thromb Vasc Biol 28(9):1596–1605. doi:10.1161/atvbaha.108.166421 PubMedCentralCrossRefPubMedGoogle Scholar
  12. Dayal S, Rodionov RN, Arning E, Bottiglieri T, Kimoto M, Murry DJ, Cooke JP, Faraci FM, Lentz SR (2008) Tissue-specific downregulation of dimethylarginine dimethylaminohydrolase in hyperhomocysteinemia. Am J Physiol Heart Circ Physiol 295(2):H816–H825. doi:10.1152/ajpheart.01348.2007 PubMedCentralCrossRefPubMedGoogle Scholar
  13. Dayoub H, Achan V, Adimoolam S, Jacobi J, Stuehlinger MC, Wang BY, Tsao PS, Kimoto M, Vallance P, Patterson AJ, Cooke JP (2003) Dimethylarginine dimethylaminohydrolase regulates nitric oxide synthesis: genetic and physiological evidence. Circulation 108(24):3042–3047. doi:10.1161/01.cir.0000101924.04515.2e CrossRefPubMedGoogle Scholar
  14. De Bree A, Verschuren WM, Kromhout D, Kluijtmans LA, Blom HJ (2002) Homocysteine determinants and the evidence to what extent homocysteine determines the risk of coronary heart disease. Pharmacol Rev 54(4):599–618CrossRefPubMedGoogle Scholar
  15. den Heijer M, Willems HP, Blom HJ, Gerrits WB, Cattaneo M, Eichinger S, Rosendaal FR, Bos GM (2007) Homocysteine lowering by B vitamins and the secondary prevention of deep vein thrombosis and pulmonary embolism: a randomized, placebo-controlled, double-blind trial. Blood 109(1):139–144. doi:10.1182/blood-2006-04-014654 CrossRefGoogle Scholar
  16. Derouiche F, Bole-Feysot C, Naimi D, Coeffier M (2014) Hyperhomocysteinemia-induced oxidative stress differentially alters proteasome composition and activities in heart and aorta. Biochem Biophys Res Commun 452(3):740–745. doi:10.1016/j.bbrc.2014.08.141 CrossRefPubMedGoogle Scholar
  17. Di Simplicio P, Frosali S, Priora R, Summa D, Cherubini Di Simplicio F, Di Giuseppe D, Di Stefano A (2005) Biochemical and biological aspects of protein thiolation in cells and plasma. Antioxid Redox Signal 7(7–8):951–963. doi:10.1089/ars.2005.7.951 CrossRefPubMedGoogle Scholar
  18. Doshi S, McDowell I, Goodfellow J, Stabler S, Böger R, Allen R, Newcombe R, Lewis M, Moat S (2005) Relationship between S-adenosylmethionine, S-adenosylhomocysteine, asymmetric dimethylarginine, and endothelial function in healthy human subjects during experimental hyper- and hypohomocysteinemia. Metabolism 54(3):351–360. doi:10.1016/j.metabol.2004.09.015 CrossRefPubMedGoogle Scholar
  19. Feng M, Whitesall S, Zhang Y, Beibel M, D’Alecy L, DiPetrillo K (2008) Validation of volume-pressure recording tail-cuff blood pressure measurements. Am J Hypertens 21(12):1288–1291. doi:10.1038/ajh.2008.301 CrossRefPubMedGoogle Scholar
  20. Fu YF, Xiong Y, Guo Z (2005) A reduction of endogenous asymmetric dimethylarginine contributes to the effect of captopril on endothelial dysfunction induced by homocysteine in rats. Eur J Pharmacol 508(1–3):167–175. doi:10.1016/j.ejphar.2004.11.063 CrossRefPubMedGoogle Scholar
  21. Gore MO, Luneburg N, Schwedhelm E, Ayers CR, Anderssohn M, Khera A, Atzler D, de Lemos JA, Grant PJ, McGuire DK, Böger RH (2013) Symmetrical dimethylarginine predicts mortality in the general population: observations from the Dallas heart study. Arterioscler Thromb Vasc Biol 33(11):2682–2688. doi:10.1161/atvbaha.113.301219 CrossRefPubMedGoogle Scholar
  22. Hanratty CG, McGrath LT, McAuley DF, Young IS, Johnston GD (2001) The effects of oral methionine and homocysteine on endothelial function. Heart 85(3):326–330PubMedCentralCrossRefPubMedGoogle Scholar
  23. Jakubowski H (2006) Pathophysiological consequences of homocysteine excess. J Nutr 136(6 Suppl):1741S–1749SPubMedGoogle Scholar
  24. Jamison RL, Hartigan P, Kaufman JS, Goldfarb DS, Warren SR, Guarino PD, Gaziano JM, Veterans Affairs Site I (2007) Effect of homocysteine lowering on mortality and vascular disease in advanced chronic kidney disease and end-stage renal disease: a randomized controlled trial. JAMA 298(10):1163–1170. doi:10.1001/jama.298.10.1163 CrossRefPubMedGoogle Scholar
  25. Lentz SR, Haynes WG (2004) Homocysteine: is it a clinically important cardiovascular risk factor? Clevel Clin J Med 71(9):729–734CrossRefGoogle Scholar
  26. Lorin J, Zeller M, Guilland JC, Cottin Y, Vergely C, Rochette L (2014) Arginine and nitric oxide synthase: regulatory mechanisms and cardiovascular aspects. Mol Nutr Food Res 58(1):101–116. doi:10.1002/mnfr.201300033 CrossRefPubMedGoogle Scholar
  27. Loscalzo J (2006) Homocysteine trials–clear outcomes for complex reasons. N Engl J Med 354(15):1629–1632. doi:10.1056/NEJMe068060 CrossRefPubMedGoogle Scholar
  28. Magne J, Huneau JF, Delemasure S, Rochette L, Tome D, Mariotti F (2009) Whole-body basal nitric oxide production is impaired in postprandial endothelial dysfunction in healthy rats. Nitric Oxide 21(1):37–43. doi:10.1016/j.niox.2009.04.003 CrossRefPubMedGoogle Scholar
  29. Mariotti F, Hammiche A, Blouet C, Dare S, Tome D, Huneau JF (2006) Medium-term methionine supplementation increases plasma homocysteine but not ADMA and improves blood pressure control in rats fed a diet rich in protein and adequate in folate and choline. Eur J Nutr 45(7):383–390. doi:10.1007/s00394-006-0611-1 CrossRefPubMedGoogle Scholar
  30. Pope AJ, Karuppiah K, Cardounel AJ (2009) Role of the PRMT-DDAH-ADMA axis in the regulation of endothelial nitric oxide production. Pharmacol Res 60(6):461–465. doi:10.1016/j.phrs.2009.07.016 PubMedCentralCrossRefPubMedGoogle Scholar
  31. Santa T, Aoyama C, Fukushima T, Imai K, Funatsu T (2006) Suppression of thiol exchange reaction in the determination of reduced-form thiols by high-performance liquid chromatography with fluorescence detection after derivatization with fluorogenic benzofurazan reagent, 7-fluoro-2,1,3-benzoxadiazole-4-sulfonate and 4-aminosulfonyl-7-fluoro-2,1,3-benzoxadiazole. Biomed Chromatogr 20(6–7):656–661. doi:10.1002/bmc.683 CrossRefPubMedGoogle Scholar
  32. Schwedhelm E, Wallaschofski H, Atzler D, Dorr M, Nauck M, Volker U, Kroemer HK, Volzke H, Böger RH, Friedrich N (2014) Incidence of all-cause and cardiovascular mortality predicted by symmetric dimethylarginine in the population-based study of health in Pomerania. PLoS One 9(5):e96875. doi:10.1371/journal.pone.0096875 PubMedCentralCrossRefPubMedGoogle Scholar
  33. Shirakawa T, Kako K, Shimada T, Nagashima Y, Nakamura A, Ishida J, Fukamizu A (2011) Production of free methylarginines via the proteasome and autophagy pathways in cultured cells. Mol Med Rep 4(4):615–620. doi:10.3892/mmr.2011.488 PubMedGoogle Scholar
  34. Siroen MP, Teerlink T, Nijveldt RJ, Prins HA, Richir MC, van Leeuwen PA (2006) The clinical significance of asymmetric dimethylarginine. Annu Rev Nutr 26:203–228. doi:10.1146/annurev.nutr.26.061505.111320 CrossRefPubMedGoogle Scholar
  35. Stuhlinger MC, Tsao PS, Her JH, Kimoto M, Balint RF, Cooke JP (2001) Homocysteine impairs the nitric oxide synthase pathway: role of asymmetric dimethylarginine. Circulation 104(21):2569–2575CrossRefPubMedGoogle Scholar
  36. Stuhlinger MC, Oka RK, Graf EE, Schmolzer I, Upson BM, Kapoor O, Szuba A, Malinow MR, Wascher TC, Pachinger O, Cooke JP (2003) Endothelial dysfunction induced by hyperhomocyst(e)inemia: role of asymmetric dimethylarginine. Circulation 108(8):933–938CrossRefPubMedGoogle Scholar
  37. Teerlink T, Nijveldt RJ, de Jong S, van Leeuwen PA (2002) Determination of arginine, asymmetric dimethylarginine, and symmetric dimethylarginine in human plasma and other biological samples by high-performance liquid chromatography. Anal Biochem 303(2):131–137. doi:10.1006/abio.2001.5575 CrossRefPubMedGoogle Scholar
  38. Teerlink T, Luo Z, Palm F, Wilcox CS (2009) Cellular ADMA: regulation and action. Pharmacol Res 60(6):448–460. doi:10.1016/j.phrs.2009.08.002 PubMedCentralCrossRefPubMedGoogle Scholar
  39. Tousoulis D, Bouras G, Antoniades C, Marinou K, Papageorgiou N, Miliou A, Hatzis G, Stefanadi E, Tsioufis C, Stefanadis C (2011) Methionine-induced homocysteinemia impairs endothelial function in hypertensives: the role of asymmetrical dimethylarginine and antioxidant vitamins. Am J Hypertens 24(8):936–942. doi:10.1038/ajh.2011.65 CrossRefPubMedGoogle Scholar
  40. Tsikas D, Böger RH, Sandmann J, Bode-Böger SM, Frölich JC (2000a) Endogenous nitric oxide synthase inhibitors are responsible for the l-arginine paradox. FEBS Lett 478(1–2):1–3CrossRefPubMedGoogle Scholar
  41. Tsikas D, Sandmann J, Savva A, Luessen P, Böger RH, Gutzki FM, Mayer B, Frölich JC (2000b) Assessment of nitric oxide synthase activity in vitro and in vivo by gas chromatography-mass spectrometry. J Chromatogr B Biomed Sci Appl 742(1):143–153CrossRefPubMedGoogle Scholar
  42. Wanby P, Brattstrom L, Brudin L, Hultberg B, Teerlink T (2003) Asymmetric dimethylarginine and total homocysteine in plasma after oral methionine loading. Scand J Clin Lab Invest 63(5):347–353CrossRefPubMedGoogle Scholar
  43. Zaidi N, Maurer A, Nieke S, Kalbacher H (2008) Cathepsin D: a cellular roadmap. Biochem Biophys Res Commun 376(1):5–9. doi:10.1016/j.bbrc.2008.08.099 CrossRefPubMedGoogle Scholar
  44. Zylberstein DE, Bengtsson C, Bjorkelund C, Landaas S, Sundh V, Thelle D, Lissner L (2004) Serum homocysteine in relation to mortality and morbidity from coronary heart disease: a 24-year follow-up of the population study of women in Gothenburg. Circulation 109(5):601–606. doi:10.1161/01.cir.0000112581.96154.ea CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag Wien 2015

Authors and Affiliations

  • Joëlle Magné
    • 1
  • Jean-François Huneau
    • 2
    • 3
  • Didier Borderie
    • 4
  • Véronique Mathé
    • 2
    • 3
  • Cécile Bos
    • 2
    • 3
  • François Mariotti
    • 2
    • 3
  1. 1.Atherosclerosis Research Unit, Department of Medicine, Center for Molecular MedicineKarolinska University Hospital, Karolinska InstitutetStockholmSweden
  2. 2.AgroParisTechCRNH-IdF, UMR914 Nutrition Physiology and Ingestive BehaviorParisFrance
  3. 3.INRACRNH-IdF, UMR914 Nutrition Physiology and Ingestive BehaviorParisFrance
  4. 4.Laboratoire de Biochimie, Groupe Hospitalier Cochin-Broca-Hôtel Dieu, Assistance Publique-Hôpitaux de Paris (AP-HP)Paris Cedex 14France

Personalised recommendations