Thromb Haemost 1997; 78(02): 827-833
DOI: 10.1055/s-0038-1657636
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Schattauer GmbH Stuttgart

Homocysteine Metabolism in Endothelial Cells of a Patient Homozygous for Cystathionine β-synthase (CS) Deficiency

E F van der Molen
1   The Department of Paediatrics, University Hospital St. Radboud, Nijmegen, The Netherlands
,
M J Hiipakka
2   Department of Molecular Biology, University of Oulu, Finland
,
H van Lith-Zanders
1   The Department of Paediatrics, University Hospital St. Radboud, Nijmegen, The Netherlands
,
G H J Boers
3   The Department of Endocrinology, University Hospital St. Radboud, Nijmegen, The Netherlands
,
L P W J van den Heuvel
1   The Department of Paediatrics, University Hospital St. Radboud, Nijmegen, The Netherlands
,
L A H Monnens
1   The Department of Paediatrics, University Hospital St. Radboud, Nijmegen, The Netherlands
,
H J Blom
1   The Department of Paediatrics, University Hospital St. Radboud, Nijmegen, The Netherlands
› Institutsangaben
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Publikationsverlauf

Received 19. 1996

Accepted after revision 08. April 1997

Publikationsdatum:
12. Juli 2018 (online)

Summary

Homocystinuria due to cystathionine β-synthase (CS) deficiency is the most common inborn error of methionine metabolism. Patients with CS-deficiency have an extremely high risk of vascular disease. The underlying mechanism is still unsolved. Dysfunction of endothelial cells could be the trigger in the formation of atherosclerosis and thrombosis. Therefore, differences in cell function were studied between normal and CS-deficient human umbilical endothelial cells (HUVECs). Total homocysteine (tHcy) concentrations in culture media as a measure of homocysteine export increased in all cell lines, including the cell line with CS-deficiency, with constant amounts of approximately 2.5 μM every 24 h. von Willebrand factor (vWF), tissue plasminogen activator (tPA) and plasminogen activator inhibitor (PAI-1) in culture media were used as markers of endothelial function and increased also with progression of culture time. The effects of additions of folate, vitamin B6 and methionine to the culture medium were studied. The homocysteine export and the markers of endothelial function did not differ between the control and the CS-deficient HUVECs under various test conditions. These data show that CS-deficient endothelial cells have normal homocysteine export and normal endothelial cell function. In CS-deficient patients the very high blood levels of homocysteine, probably due to deficient CS function in liver and kidney, seems to be the hazardous factor to endothelial cells, thus promoting atherosclerosis and thrombosis in CS-deficient patients.

 
  • References

  • 1 Mudd SH, Levy HL, Skovby F. Disorders of transsulfuration. In: The metabolic basis of inherited disease. Scriver CR, Beaudet AL, Sly WS. et al. (eds). New York: Mc-Graw-Hill; 1995. pp 1279-1327
  • 2 Kluijtmans LAJ, Van denHeuvel LPWJ, Boers GHJ, Stevens EMB, Frosst P, Van OostBA, Trijbels FJM, Rozen R, Blom HJ. Molecular genetic analysis in mild hyperhomocysteinemia: a common mutation in the methylene-tetrahydrofolate reductase gene is a genetic risk factor for cardiovascular disease. Am J Hum Genet 1996; 58: 35-41
  • 3 Kraus JP. Molecular basis of phenotype expression in homocystinuria. J Inher Metab Dis 1994; 17: 383-390
  • 4 Loscalzo J. The oxidant stress of hyperhomocysteinemia. J Clin Invest 1996; 98 (01) 5-7
  • 5 Starkebaum G, Harlan JM. Endothelial cell injury due to copper-catalyzed hydrogen peroxide generation from homocysteine. J Clin Invest 1986; 77: 1370-1376
  • 6 Blom HJ, Kleinveld HA, Boers GHJ, Demacker PNM, Hak-Lemmers HLM, Te Poele-PothoffMTWB, Trijbels FJM. Lipid peroxidation and susceptibility of low-density lipoprotein to in-vitro oxidation in hyperhomocysteinemia. Eur J Clin Invest 1995; 25: 149-154
  • 7 Dudman NPB, Wilcken DEL, Stocker R. Circulating lipid hydroxy peroxide levels in human hyperhomocysteinemia. Arteriosclerosis and thrombosis 1993; 13: 512-516
  • 8 Stehouwer CDA, Nauta JJP, Zeldenrust GC, Hackeng WHL, Donker AJM, Den OttolanderGHJ. Urinary albumin secretion, cardiovascular disease, and endothelial dysfunction in non-insulin-dependent diabetes mellitus. Lancet 1992; 340: 319-323
  • 9 Ridker PM, Vaughan DE, Stampfer MJ, Manson JE, Hennekes CH. Endogenous tissue-type plasminogen activator and risk of myocardial infarction. Lancet 1993; 341: 1165-1168
  • 10 Pandolfi M. The role of fibrinolytic factors in ischaemia. Eye 1991; 05: 159-169
  • 11 Lentz SR, Sadler JE. Homocysteine inhibits von Willebrand factor processing and secretion by preventing transport from the endoplasmic reticulum. Blood 1993; 81 (03) 683-689
  • 12 Rodgers GM, Conn MT. Homocysteine, an atherogenic stimulus, reduces protein-C activation by arterial and venous endothelial cells. Blood 1990; 75: 895-901
  • 13 Nishinaga M, Ozawa T, Shimada K. Homocysteine, a thrombogenic agent, suppresses anticoagulant heparan sulphate expression in cultured porcine aortic endothelial cells. J Clin Invest 1993; 92: 1381-1386
  • 14 Hajjar KA. Homocysteine-induced modulation of tissue plasminogen activator binding to its endothelial cell membrane receptor. J Clin Invest 1993; 91: 2873-2879
  • 15 Blom HJ, van derMolen EF. Pathobiochemical implications of hyperhomocysteinemia. Fibrinolysis 1994; 08: 86-87
  • 16 Refsum H, Christensen B, Djurhuus R, Ueland PM. Interaction between methotrexate,“rescue” agents and cell proliferation as modulators of homocysteine export from cells in culture. J Pharmacol Exp Ther 1991; 02 258 559-566
  • 17 Christensen B, Refsum H, Garras A, Ueland PM. Homocysteine remethylation during nitrous oxide exposure of cells cultured in media containing various concentrations of folates. J Phar Exp Therap 1992; 261 (03) 1096-1105
  • 18 Christensen B, Rosenblatt DS, Chu RC, Ueland PM. Effect of methionine and nitrous oxide on homocysteine export and remethylation in fibroblasts from cystathionine synthase-deficient cblG and cblE patients. Paediatric Research 1993; 35 (01) 3-9
  • 19 Ueland PM, Refsum H, Brattström L. Plasma homocysteine and vascular disease. In: Atherosclerotic Cardiovascular Disease, Haemostasis and Endothelial Function. Francis RB. (ed). New York: Marcel Dekker Inc; 1992. pp 183-236
  • 20 Van derMolen EF, Van denHeuvel LPWJ, Te Poele-PothoffMTWB, Monnens LAH, Eskes TKAB, Blom HJ. The effect of folic acid on the homocysteine metabolism in human umbilical vein endothelial cells (HUVECs). Eur J Clin Invest 1996; 26: 304-309
  • 21 Jaffe E, Nachmann RL, Becker CG, Minick CR. Culture of human endothelial cell derived from umbilical veins. Identification by morphologic and immunologic criteria. J Clin Invest 1973; 52: 2745
  • 22 Oliver MH, Harrison NK, Bishop JE, Cole PJ, Laurent GJ. A rapid and convenient assay for counting cells cultured in microwell plates: application for assessment of growth factors. J Cell Science 1989; 92: 513-518
  • 23 Te Poele-PothoffMTWB, Van derBerg M, Franken DG, Boers GHJ, Jacobs C, de KroonM, Eskes TKAB, Trijbels FJM, Blom HJ. Three different methods for the determination of total homocysteine in plasma. Ann Clin Biochem 1995; 32: 218-220
  • 24 Fowler B, Krus J, Packman S, Rosenberg LE. Homocystinuria. Evidence of three distinct classes of cystathionine p-synthase mutants in cultured fibroblasts. J Clin Invest 1978; 61: 645-653
  • 25 Lowry OH, Rosebrouhg NJ, Farr AL, Randall RJ. Protein measurement with the folin phenol reagent. J Biol Chem 1951; 193: 265-275
  • 26 Ingerslev JA. Sensitive ELISA for von Willebrand Factor (vWF'.Ag). Scand J Clin Lab Invest 1987; 47: 143-149
  • 27 Finkelstein JD. Methionine metabolism in mammals. J Nutr 1990; 1: 228-237
  • 28 Van HinsberghVWM. Regulation of the synthesis and secretion of Plasminogen Activators by endothelial cells. Haemostasis 1988; 18: 307-327
  • 29 Schleef RR, Loskutoff DJ. Fibrinolytic system of vascular endothelial cells. Haemostasis 1988; 18: 328-341
  • 30 Paleolog EM, Crossman DC, Mcvey JH, Pearson JD. Differential regulation by cytokines of constitutive and stimulated secretion of von Willebrand Factor from endothelial cells. Blood 1990; 75 (03) 688-695
  • 31 De GrootPG, Willems C, Boers GHJ, Gonsalves MD, Van AkenWG, Van MourikJA. Endothelial cell dysfunction in homocystinuria. Eur J Clin Invest 1983; 13: 405-410
  • 32 De ValkHW, Van EedenMKG, Banga JD, Van derGriend R, De GrootE, Haas FJLM, Meuwissen OJAT, Duran M, Smeitink JAM, Bwee TienPoll-The, De KlerkJBC. Evaluation of the presence of premature atherosclerosis in adults with heterozygosity for cystathionine ²-synthase deficiency. Stroke 1996; 27 (06) 1134-1136
  • 33 Frosst P, Blom HJ, Goyette P, Sheppard CA, Matthews RG, Boers GHJ, Den HeijerM, Kluijtmans LAJ, Van denHeuvel LPWJ, Rozen R. A candidate genetic risk factor for vascular disease: a common mutation in methylenetetrahydrofolate reductase. Nat Genet 1995; 10: 111-113