Logo of biochemjBJ Latest papers and much more!
Biochem J. 1998 May 15; 332(Pt 1): 213–221.
PMCID: PMC1219470

Characterization of the stress-inducing effects of homocysteine.


The mechanism by which homocysteine causes endothelial cell (EC) injury and/or dysfunction is not fully understood. To examine the stress-inducing effects of homocysteine on ECs, mRNA differential display and cDNA microarrays were used to evaluate changes in gene expression in cultured human umbilical-vein endothelial cells (HUVEC) exposed to homocysteine. Here we show that homocysteine increases the expression of GRP78 and GADD153, stress-response genes induced by agents or conditions that adversely affect the function of the endoplasmic reticulum (ER). Induction of GRP78 was specific for homocysteine because other thiol-containing amino acids, heat shock or H2O2 did not appreciably increase GRP78 mRNA levels. Homocysteine failed to elicit an oxidative stress response in HUVEC because it had no effect on the expression of heat shock proteins (HSPs) including HSP70, nor did it activate heat shock transcription factor 1. Furthermore homocysteine blocked the H2O2-induced expression of HSP70. In support of our findings in vitro, steady-state mRNA levels of GRP78, but not HSP70, were elevated in the livers of cystathionine beta-synthase-deficient mice with hyperhomocysteinaemia. These studies indicate that the activation of stress response genes by homocysteine involves reductive stress leading to altered ER function and is in contrast with that of most other EC perturbants. The observation that homocysteine also decreases the expression of the antioxidant enzymes glutathione peroxidase and natural killer-enhancing factor B suggests that homocysteine could potentially enhance the cytotoxic effect of agents or conditions known to cause oxidative stress.

Full Text

The Full Text of this article is available as a PDF (410K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Ross R. The pathogenesis of atherosclerosis: a perspective for the 1990s. Nature. 1993 Apr 29;362(6423):801–809. [PubMed]
  • Navab M, Berliner JA, Watson AD, Hama SY, Territo MC, Lusis AJ, Shih DM, Van Lenten BJ, Frank JS, Demer LL, et al. The Yin and Yang of oxidation in the development of the fatty streak. A review based on the 1994 George Lyman Duff Memorial Lecture. Arterioscler Thromb Vasc Biol. 1996 Jul;16(7):831–842. [PubMed]
  • Navab M, Fogelman AM, Berliner JA, Territo MC, Demer LL, Frank JS, Watson AD, Edwards PA, Lusis AJ. Pathogenesis of atherosclerosis. Am J Cardiol. 1995 Sep 28;76(9):18C–23C. [PubMed]
  • Lüscher TF, Tanner FC, Tschudi MR, Noll G. Endothelial dysfunction in coronary artery disease. Annu Rev Med. 1993;44:395–418. [PubMed]
  • Zhu W, Roma P, Pellegatta F, Catapano AL. Oxidized-LDL induce the expression of heat shock protein 70 in human endothelial cells. Biochem Biophys Res Commun. 1994 Apr 15;200(1):389–394. [PubMed]
  • Zhu W, Roma P, Pirillo A, Pellegatta F, Catapano AL. Human endothelial cells exposed to oxidized LDL express hsp70 only when proliferating. Arterioscler Thromb Vasc Biol. 1996 Sep;16(9):1104–1111. [PubMed]
  • Jornot L, Mirault ME, Junod AF. Differential expression of hsp70 stress proteins in human endothelial cells exposed to heat shock and hydrogen peroxide. Am J Respir Cell Mol Biol. 1991 Sep;5(3):265–275. [PubMed]
  • Yellon DM, Latchman DS. Stress proteins and myocardial protection. J Mol Cell Cardiol. 1992 Feb;24(2):113–124. [PubMed]
  • Hightower LE. Heat shock, stress proteins, chaperones, and proteotoxicity. Cell. 1991 Jul 26;66(2):191–197. [PubMed]
  • Georgopoulos C, Welch WJ. Role of the major heat shock proteins as molecular chaperones. Annu Rev Cell Biol. 1993;9:601–634. [PubMed]
  • Kim AT, Sarafian TA, Shau H. Characterization of antioxidant properties of natural killer-enhancing factor-B and induction of its expression by hydrogen peroxide. Toxicol Appl Pharmacol. 1997 Nov;147(1):135–142. [PubMed]
  • Pinkus R, Weiner LM, Daniel V. Role of quinone-mediated generation of hydroxyl radicals in the induction of glutathione S-transferase gene expression. Biochemistry. 1995 Jan 10;34(1):81–88. [PubMed]
  • Lautier D, Luscher P, Tyrrell RM. Endogenous glutathione levels modulate both constitutive and UVA radiation/hydrogen peroxide inducible expression of the human heme oxygenase gene. Carcinogenesis. 1992 Feb;13(2):227–232. [PubMed]
  • Kang SS, Wong PW, Malinow MR. Hyperhomocyst(e)inemia as a risk factor for occlusive vascular disease. Annu Rev Nutr. 1992;12:279–298. [PubMed]
  • Selhub J, Jacques PF, Bostom AG, D'Agostino RB, Wilson PW, Belanger AJ, O'Leary DH, Wolf PA, Schaefer EJ, Rosenberg IH. Association between plasma homocysteine concentrations and extracranial carotid-artery stenosis. N Engl J Med. 1995 Feb 2;332(5):286–291. [PubMed]
  • Mayer EL, Jacobsen DW, Robinson K. Homocysteine and coronary atherosclerosis. J Am Coll Cardiol. 1996 Mar 1;27(3):517–527. [PubMed]
  • McCully KS. Homocysteine and vascular disease. Nat Med. 1996 Apr;2(4):386–389. [PubMed]
  • Harker LA, Slichter SJ, Scott CR, Ross R. Homocystinemia. Vascular injury and arterial thrombosis. N Engl J Med. 1974 Sep 12;291(11):537–543. [PubMed]
  • Harker LA, Harlan JM, Ross R. Effect of sulfinpyrazone on homocysteine-induced endothelial injury and arteriosclerosis in baboons. Circ Res. 1983 Dec;53(6):731–739. [PubMed]
  • Hladovec J. Experimental homocystinemia, endothelial lesions and thrombosis. Blood Vessels. 1979;16(4):202–205. [PubMed]
  • Wall RT, Harlan JM, Harker LA, Striker GE. Homocysteine-induced endothelial cell injury in vitro: a model for the study of vascular injury. Thromb Res. 1980 Apr 1;18(1-2):113–121. [PubMed]
  • Starkebaum G, Harlan JM. Endothelial cell injury due to copper-catalyzed hydrogen peroxide generation from homocysteine. J Clin Invest. 1986 Apr;77(4):1370–1376. [PMC free article] [PubMed]
  • de Groot PG, Willems C, Boers GH, Gonsalves MD, van Aken WG, van Mourik JA. Endothelial cell dysfunction in homocystinuria. Eur J Clin Invest. 1983 Oct;13(5):405–410. [PubMed]
  • Dudman NP, Hicks C, Wang J, Wilcken DE. Human arterial endothelial cell detachment in vitro: its promotion by homocysteine and cysteine. Atherosclerosis. 1991 Nov;91(1-2):77–83. [PubMed]
  • Fryer RH, Wilson BD, Gubler DB, Fitzgerald LA, Rodgers GM. Homocysteine, a risk factor for premature vascular disease and thrombosis, induces tissue factor activity in endothelial cells. Arterioscler Thromb. 1993 Sep;13(9):1327–1333. [PubMed]
  • Rodgers GM, Conn MT. Homocysteine, an atherogenic stimulus, reduces protein C activation by arterial and venous endothelial cells. Blood. 1990 Feb 15;75(4):895–901. [PubMed]
  • Lentz SR, Sadler JE. Inhibition of thrombomodulin surface expression and protein C activation by the thrombogenic agent homocysteine. J Clin Invest. 1991 Dec;88(6):1906–1914. [PMC free article] [PubMed]
  • Rodgers GM, Kane WH. Activation of endogenous factor V by a homocysteine-induced vascular endothelial cell activator. J Clin Invest. 1986 Jun;77(6):1909–1916. [PMC free article] [PubMed]
  • Hajjar KA. Homocysteine-induced modulation of tissue plasminogen activator binding to its endothelial cell membrane receptor. J Clin Invest. 1993 Jun;91(6):2873–2879. [PMC free article] [PubMed]
  • Stamler JS, Osborne JA, Jaraki O, Rabbani LE, Mullins M, Singel D, Loscalzo J. Adverse vascular effects of homocysteine are modulated by endothelium-derived relaxing factor and related oxides of nitrogen. J Clin Invest. 1993 Jan;91(1):308–318. [PMC free article] [PubMed]
  • Lentz SR, Sobey CG, Piegors DJ, Bhopatkar MY, Faraci FM, Malinow MR, Heistad DD. Vascular dysfunction in monkeys with diet-induced hyperhomocyst(e)inemia. J Clin Invest. 1996 Jul 1;98(1):24–29. [PMC free article] [PubMed]
  • Rolland PH, Friggi A, Barlatier A, Piquet P, Latrille V, Faye MM, Guillou J, Charpiot P, Bodard H, Ghiringhelli O, et al. Hyperhomocysteinemia-induced vascular damage in the minipig. Captopril-hydrochlorothiazide combination prevents elastic alterations. Circulation. 1995 Feb 15;91(4):1161–1174. [PubMed]
  • Loscalzo J. The oxidant stress of hyperhomocyst(e)inemia. J Clin Invest. 1996 Jul 1;98(1):5–7. [PMC free article] [PubMed]
  • Kokame K, Kato H, Miyata T. Homocysteine-respondent genes in vascular endothelial cells identified by differential display analysis. GRP78/BiP and novel genes. J Biol Chem. 1996 Nov 22;271(47):29659–29665. [PubMed]
  • Pelham HR. Speculations on the functions of the major heat shock and glucose-regulated proteins. Cell. 1986 Sep 26;46(7):959–961. [PubMed]
  • Jaffe EA, Nachman RL, Becker CG, Minick CR. Culture of human endothelial cells derived from umbilical veins. Identification by morphologic and immunologic criteria. J Clin Invest. 1973 Nov;52(11):2745–2756. [PMC free article] [PubMed]
  • Kassis J, Hirsh J, Podor TJ. Evidence that postoperative fibrinolytic shutdown is mediated by plasma factors that stimulate endothelial cell type I plasminogen activator inhibitor biosynthesis. Blood. 1992 Oct 1;80(7):1758–1764. [PubMed]
  • Watanabe M, Osada J, Aratani Y, Kluckman K, Reddick R, Malinow MR, Maeda N. Mice deficient in cystathionine beta-synthase: animal models for mild and severe homocyst(e)inemia. Proc Natl Acad Sci U S A. 1995 Feb 28;92(5):1585–1589. [PMC free article] [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • Austin RC, Howard PL, D'Souza VN, Klamut HJ, Ray PN. Cloning and characterization of alternatively spliced isoforms of Dp71. Hum Mol Genet. 1995 Sep;4(9):1475–1483. [PubMed]
  • Hill SA, Shaughnessy SG, Joshua P, Ribau J, Austin RC, Podor TJ. Differential mechanisms targeting type 1 plasminogen activator inhibitor and vitronectin into the storage granules of a human megakaryocytic cell line. Blood. 1996 Jun 15;87(12):5061–5073. [PubMed]
  • Sarge KD, Murphy SP, Morimoto RI. Activation of heat shock gene transcription by heat shock factor 1 involves oligomerization, acquisition of DNA-binding activity, and nuclear localization and can occur in the absence of stress. Mol Cell Biol. 1993 Mar;13(3):1392–1407. [PMC free article] [PubMed]
  • Fortin LJ, Genest J., Jr Measurement of homocyst(e)ine in the prediction of arteriosclerosis. Clin Biochem. 1995 Apr;28(2):155–162. [PubMed]
  • Liang P, Pardee AB. Differential display of eukaryotic messenger RNA by means of the polymerase chain reaction. Science. 1992 Aug 14;257(5072):967–971. [PubMed]
  • Huang LE, Zhang H, Bae SW, Liu AY. Thiol reducing reagents inhibit the heat shock response. Involvement of a redox mechanism in the heat shock signal transduction pathway. J Biol Chem. 1994 Dec 2;269(48):30718–30725. [PubMed]
  • Braakman I, Helenius J, Helenius A. Role of ATP and disulphide bonds during protein folding in the endoplasmic reticulum. Nature. 1992 Mar 19;356(6366):260–262. [PubMed]
  • Graven KK, Zimmerman LH, Dickson EW, Weinhouse GL, Farber HW. Endothelial cell hypoxia associated proteins are cell and stress specific. J Cell Physiol. 1993 Dec;157(3):544–554. [PubMed]
  • Watowich SS, Morimoto RI. Complex regulation of heat shock- and glucose-responsive genes in human cells. Mol Cell Biol. 1988 Jan;8(1):393–405. [PMC free article] [PubMed]
  • Jornot L, Mirault ME, Junod AF. Differential expression of hsp70 stress proteins in human endothelial cells exposed to heat shock and hydrogen peroxide. Am J Respir Cell Mol Biol. 1991 Sep;5(3):265–275. [PubMed]
  • Lu D, Maulik N, Moraru II, Kreutzer DL, Das DK. Molecular adaptation of vascular endothelial cells to oxidative stress. Am J Physiol. 1993 Mar;264(3 Pt 1):C715–C722. [PubMed]
  • Halleck MM, Holbrook NJ, Skinner J, Liu H, Stevens JL. The molecular response to reductive stress in LLC-PK1 renal epithelial cells: coordinate transcriptional regulation of gadd153 and grp78 genes by thiols. Cell Stress Chaperones. 1997 Mar;2(1):31–40. [PMC free article] [PubMed]
  • Lentz SR, Sadler JE. Homocysteine inhibits von Willebrand factor processing and secretion by preventing transport from the endoplasmic reticulum. Blood. 1993 Feb 1;81(3):683–689. [PubMed]
  • Chen Q, Yu K, Holbrook NJ, Stevens JL. Activation of the growth arrest and DNA damage-inducible gene gadd 153 by nephrotoxic cysteine conjugates and dithiothreitol. J Biol Chem. 1992 Apr 25;267(12):8207–8212. [PubMed]
  • Wang XZ, Lawson B, Brewer JW, Zinszner H, Sanjay A, Mi LJ, Boorstein R, Kreibich G, Hendershot LM, Ron D. Signals from the stressed endoplasmic reticulum induce C/EBP-homologous protein (CHOP/GADD153). Mol Cell Biol. 1996 Aug;16(8):4273–4280. [PMC free article] [PubMed]
  • Whelan SA, Hightower LE. Differential induction of glucose-regulated and heat shock proteins: effects of pH and sulfhydryl-reducing agents on chicken embryo cells. J Cell Physiol. 1985 Nov;125(2):251–258. [PubMed]
  • Tsai JC, Perrella MA, Yoshizumi M, Hsieh CM, Haber E, Schlegel R, Lee ME. Promotion of vascular smooth muscle cell growth by homocysteine: a link to atherosclerosis. Proc Natl Acad Sci U S A. 1994 Jul 5;91(14):6369–6373. [PMC free article] [PubMed]
  • Loven DP. A role for reduced oxygen species in heat induced cell killing and the induction of thermotolerance. Med Hypotheses. 1988 May;26(1):39–50. [PubMed]
  • Morris JA, Dorner AJ, Edwards CA, Hendershot LM, Kaufman RJ. Immunoglobulin binding protein (BiP) function is required to protect cells from endoplasmic reticulum stress but is not required for the secretion of selective proteins. J Biol Chem. 1997 Feb 14;272(7):4327–4334. [PubMed]
  • Liu H, Bowes RC, 3rd, van de Water B, Sillence C, Nagelkerke JF, Stevens JL. Endoplasmic reticulum chaperones GRP78 and calreticulin prevent oxidative stress, Ca2+ disturbances, and cell death in renal epithelial cells. J Biol Chem. 1997 Aug 29;272(35):21751–21759. [PubMed]
  • Dorner AJ, Wasley LC, Kaufman RJ. Overexpression of GRP78 mitigates stress induction of glucose regulated proteins and blocks secretion of selective proteins in Chinese hamster ovary cells. EMBO J. 1992 Apr;11(4):1563–1571. [PMC free article] [PubMed]
  • Dorner AJ, Wasley LC, Kaufman RJ. Protein dissociation from GRP78 and secretion are blocked by depletion of cellular ATP levels. Proc Natl Acad Sci U S A. 1990 Oct;87(19):7429–7432. [PMC free article] [PubMed]
  • Upchurch GR, Jr, Welch GN, Fabian AJ, Freedman JE, Johnson JL, Keaney JF, Jr, Loscalzo J. Homocyst(e)ine decreases bioavailable nitric oxide by a mechanism involving glutathione peroxidase. J Biol Chem. 1997 Jul 4;272(27):17012–17017. [PubMed]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society


Save items

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


  • Compound
    PubChem Compound links
  • MedGen
    Related information in MedGen
  • PubMed
    PubMed citations for these articles
  • Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...