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Vitamin C promotes human endothelial cell growth via the ERK-signaling pathway

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Abstract

Background

Epidemiological, secondary prevention and small interventional trials suggest a preventive role of vitamin C for cardiovascular diseases (CAD), especially through improving endothelial dysfunction. Large primary prevention trials failed to confirm this. Mechanistic studies may contribute to resolve this discrepancy.

Aim of the study

We examined whether vitamin C activates mitogen-activated protein kinases (MAPK) in human umbilical cord venous endothelial cells (HUVECs) and whether reactive oxygen species (ROS) play a role in this process.

Methods

Subconfluent quiescent HUVECs were incubated with vitamin C alone or in combination with catalase (CAT) and/or hydrogenperoxide (H2O2). Intracellular MAPK were determined by Western blot, proliferation by cell count and DNA-synthesis by [3H]-thymidine-uptake.

Results

HUVECs were incubated with vitamin C (60 µM) for 5–60 min or for 20 min (30–90 µM). A dose-dependent phosphorylation of extracellular signal-regulated-kinases (ERKs)-1 and -2 with a maximum of phosphorylation at 15–20 min was observed and inhibitable by MEK1/2-inhibitor U0126 (5–10 µM). Vitamin C (60 µM) stimulated phosphorylation of ERK5, but not of p38 and c-Jun, demonstrating a different MAPK-activation pattern compared to H2O2. Vitamin C (60 µM) induced proliferation and a dose-dependent [3H]-thymidine-uptake (30–120 µM) within 20 h. CAT (0.3 U/ml) did neither suppress the vitamin C induced [3H]-thymidine-uptake nor ERK1/2-phosphorylation. CAT (0.3 U/ml), but not vitamin C (60 µM) abrogated the inhibitory effects of H2O2 (100 µM) on [3H]-thymidine-uptake.

Conclusion

Physiological vitamin C-concentrations promote proliferation of subconfluent ECs by activating an ERK1/2 controlled pathway. Targeting MAPK by vitamin C may improve, besides antioxidant mechanisms, endothelial dysfunction by promoting a fast regeneration of the endothelium after tissue injury, particularly required during secondary prevention and early development.

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References

  1. Kris-Etherton PM, Lichtenstein AH, Howard BV, Steinberg D, Witztum JL, Nutrition Committee of the American Heart Association Council on Nutrition, Physical Activity, Metabolism (2004) Antioxidant vitamin supplements and cardiovascular disease. Circulation 110(5):637–641

    Article  CAS  Google Scholar 

  2. Chen WT, Yan HC, Yu FC (2003) Vitamin C improves vascular resistence in patients with chronic renal failure. Kidney Int 64:2325–2326

    Article  Google Scholar 

  3. Nyyssönen K, Poulsen HE, Hayn M, Agerbo P, Porkkala-Sarataho E, Kaikkonen J, Salonen R, Salonen JT (1997) Effect of supplementation of smoking men with plain or slow release ascorbic acid on lipoprotein oxidation. Eur J Clin Nutr 51:154–163

    Article  Google Scholar 

  4. Ulrich-Merzenich G, Metzner C, Bhonde RR, Malsch G, Schiermeyer B, Vetter H (2002) Simultaneous isolation of endothelial and smooth muscle cells from human umbilical artery or vein and their growth response to low-density lipoproteins. In Vitro Cell Dev Biol Anim 38(5):265–272

    Article  Google Scholar 

  5. Totzke G, Metzner C, Ulrich-Merzenich G, Ko Y, Sachinidis A, Vetter H (2001) Effect of vitamin E and vitamin C on the DNA-synthesis of human umbilical arterial endothelial cells. Eur J Nutr 40(3):121–126

    Article  CAS  Google Scholar 

  6. Napoli C, de Nigris F, Palinski W (2001) Multiple role of reactive oxygen species in the arterial wall. J Cell Biochem 82:674–682

    Article  CAS  Google Scholar 

  7. Schwartz SM, Gajdusek CM, Selden SC 3rd (1981) Vascular wall growth control: the role of the endothelium. Arteriosclerosis 1(2):107–126

    Google Scholar 

  8. Napoli C, Cicala C, Wallace JL, de Nigris F, Santagada V, Caliendo G, Franconi F, Inarro LJ, Cirino G (2000) Protease-activated receptor-2 modulates myocardial ischemia-reperfusion injury in the rat heart. PNAS 97:3678–3683

    Article  CAS  Google Scholar 

  9. Hanahan D, Folkman J. (1996) Patterns of emerging mechanisms of the angiogenic switch during tumorgenesis. Cell 86:353–364

    Article  CAS  Google Scholar 

  10. Reidy MA, Schwartz SM. (1981) Endothelial regeneration. III. Time course of intimal changes after small defined injury to rat aortic endothelium. Lab Invest 44(4):301–308

    CAS  Google Scholar 

  11. Nunes GL, Robinson K, Kalynych A, King III SB, Sgoutas DS, Berk BC (1997) Vitamins C and E inhibit superoxidradical production in the pig coronary artery. Circulation 96:3593–3601

    CAS  Google Scholar 

  12. Ulrich-Merzenich G, Metzner C, Schiermeyer B, Vetter H (2002) Vitamin C and vitamin E antagonistically modulate human vascular endothelial and smooth muscle cell DNA-synthesis and proliferation. Eur J Nutr 41:27–34

    Article  CAS  Google Scholar 

  13. Kyriakis JM, Avruch J (2001) Mammalian mitogen-activated protein kinase signal transduction pathways activated by stress and inflammation. Physiol Rev 81(2):807–869

    CAS  Google Scholar 

  14. Liu W, Schoenkermann A, Lowe WL Jr (2000) Activation of members of the mitogen-activated protein kinase family by glucose in endothelial cells. Am J Physiol Endocrinol Metab 279(4):E782–E790

    CAS  Google Scholar 

  15. Kyriakis JM, Banerjee P, Nikolakaki E, Dai T, Rubie EA, Ahmad MF, Avruch J, Woodgett JR (1994) The stress-activated protein kinase subfamily of c-Jun kinases. Nature 369:156–160

    Article  CAS  Google Scholar 

  16. Gouni-Berthold I, Seul C, Ko Y, Hescheler J, Sachinidis A (2001) Gangliosides GM1 and GM2 induce vascular smooth muscle cell proliferation via extracellular signal-regulated kinase 1/2 pathway. Hypertension 38:1030–1037

    CAS  Google Scholar 

  17. Bokemeyer D, Panek D, Lindemann M, Katahara M, Boor P, Kerjaschki D, Trzaskos JM, Floege J, Kramer HJ, Ostendorf T (2002) In vivo identification of the MAP-kinase cascade as a central pathogenic pathway in experimental mesangioproliferative glomerulonephritis. J Am Soc Nephrol 13(6):1473–1480

    Article  CAS  Google Scholar 

  18. Brand A, Gil S, Seger R, Yavin E (2001) Lipid constituents in oligodendroglial cells alter susceptibility to H2O2- induced apoptotic cell death via ERK activation. J Neurochem 76:910–918

    Article  CAS  Google Scholar 

  19. Kurl S, Tuomainen TP, Laukkanen JA, Nyyssonen K, Lakka T, Sivenius J, Salonen JT (2002) Plasma vitamin C modifies the association between hypertension and risk of stroke. Stroke 33(6):1568–1573

    Article  CAS  Google Scholar 

  20. Heseker H, Schneider R, Moch KJ, Kohlmeier M, Kübler W (1992) Vitaminversorgung Erwachsener in der Bundesrepublik Deutschland, VERA-Schriftenreihe Bd. IV. Wiss. Fachverlag Dr. Fieck, Niederkleen

  21. Vitamin C. In: Deutsche Gesellschaft für Ernährung (DGE); Österreichische Gesellschaft für Ernährung (ÖGE), Schweizerische Gesellschaft für Ernährungsforschung (SGE), Schweizerische Gesellschaft für Ernährung (SVE) (2000) Referenzwerte für die Nährstoffzufuhr. Umschau Braus GmbH Verlagsgesellschaft, Frankfurt am Main, pp 137–44

  22. Levine M, Dariwal KR, Welch RW, Wang Y, Park JB (1995) Determination of optimal Vitamin C requirements in humans. Am J Clin Nutr 62(6Suppl):1347S–1356S

    CAS  Google Scholar 

  23. May JM, Qu ZC (2005) Transport and intracellular accumulation of vitamin C in endothelial cells: relevance to collagen synthesis. Arch Biochem Biophys 434(1):178–186

    Article  CAS  Google Scholar 

  24. Langlois M, Duprez D, Dlanghe J, De Buyzere M, Clement D (2001) Serum vitamin C concentration is low in peripheral arterial disease and is associated with inflammation and severity of atherosclerosis. Circulation 103:1863

    CAS  Google Scholar 

  25. Riemersma RA, Carruthers F, Elton RA, Fox KAA (2000) Vitamin C and the risk of acute myocardial infarction. Am J Clin Nutr 71:1181–1186

    CAS  Google Scholar 

  26. Naidu KA (2003). Vitamin C in human health and disease is still a mistery? An overview. Nutr J 2:7

    Article  Google Scholar 

  27. Alper G, Olukman M, Irer S, Caglayan O, duman E, Yilmaz C, Ulker S (2006) Effect of vitamin E and C supplementation combined with oral antidiabetic therapy on the endothelial dysfunction in the neonatally streptozotocin injected diabetic rat. Diabetes Metab Res Rev Oct 22(3):190–197

    Article  CAS  Google Scholar 

  28. Rhee SG (1999) Redox signaling: hydrogen peroxide as intracellular messenger. Exp Mol Med 31:53–59

    CAS  Google Scholar 

  29. Lin SJ, Shyue SK, Liu PL, Chen YH, Ku HH, Chen JW, Tam KB, Chen YL (2004) Adenovirus-mediated overexpression of catalase attenuates oxLDL-induced apoptosis in human aortic endothelial cells via AP-1 and C-Jun N-terminal kinase/extracellular signalregulated kinase mitogen-activated protein kinase pathways. J Mol Cell Cardiol 36(1):129–139

    Article  CAS  Google Scholar 

  30. Bowie AG, O’Neil LA. (2000) Vitamin C inhibits NF-kappa B activation by TNF via the activation of p38 mitogen-activated protein kinase. J Immunol 165(12):7180–7188

    CAS  Google Scholar 

  31. Shi Y, Gaestel M (2002) In the cellular garden of forking paths: how p38 MAPKs signal for downstream assistance. Biol Chem 383(10):1519–1536

    Article  CAS  Google Scholar 

  32. Regan CP, Li W, Boucher DM, Spatz S, Su MS, Kuida K (2002) Erk5 null mice display multiple extraembryonic vascular and embryonic cardiovascular defects. Proc Natl Acad Sci USA 99(14):9248–9253

    Article  CAS  Google Scholar 

  33. Sohn SJ, Sarvis BK, Cado D, Winoto A (2002) ERK5 MAPK regulates embryonic angiogenesis and acts as a hypoxia-sensitive repressor vascular endothelial growth factor expression. J Biol Chem 277(45):43344–43351

    Article  CAS  Google Scholar 

  34. Pi X, Yan C, Berk BC (2004) Big Mitogen-activated protein kinase (BMK1)/ERK5 protects endothelial cells from apoptosis. Cir Res 94(3):362–369

    Article  CAS  Google Scholar 

  35. Yue TL, Wang C, Gu JL, Ma XL, Kumar S, Lee JC, Feuerstein GZ, Thomas H, Maleeff B, Ohlstein EH (2000) Inhibition of extracellular regulated kinase enhances ischemia/reoxygenation reperfusion-induced apoptosis in cultured cardiac myocytes and exaggerates reperfusion injury in isolated perfused heart. Circ Res 86:692–699

    CAS  Google Scholar 

  36. Ravingerova T, Barancik M, Strniskova M (2003) Mitogen-activated protein kinases: a new therapeutic target in cardiac pathology. Mol Cell Biochem 247(1–2):127–138

    Article  CAS  Google Scholar 

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Acknowledgements

Authors would like to thank Frederik Hartbrod for excellent technical assistance and Gerhard Malsch for the proliferation assays.

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Authors and Affiliations

  1. Medical Policlinic of the Rheinische Friedrich-Wilhelms-University of Bonn, Wilhelmstr. 35-37, 53111, Bonn, Germany

    Gudrun Ulrich-Merzenich, Heike Zeitler, Darius Panek & Hans Vetter

  2. Augusta-Krankenanstalten, Bochum, Germany

    Dirk Bokemeyer

Authors
  1. Gudrun Ulrich-Merzenich
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  2. Heike Zeitler
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  3. Darius Panek
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  4. Dirk Bokemeyer
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  5. Hans Vetter
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Corresponding author

Correspondence to Gudrun Ulrich-Merzenich.

Additional information

G. Ulrich-Merzenich and H. Zeitler are contributed equally to this work.

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Ulrich-Merzenich, G., Zeitler, H., Panek, D. et al. Vitamin C promotes human endothelial cell growth via the ERK-signaling pathway. Eur J Nutr 46, 87–94 (2007). https://doi.org/10.1007/s00394-006-0636-5

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  • DOI: https://doi.org/10.1007/s00394-006-0636-5

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