title: Characterization of the stress-inducing effects of homocysteine.
abstract: 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.
authors.affiliation: Departments of Pathology and Medicine, McMaster University and the Hamilton Civic Hospitals Research Centre, Hamilton, Ontario, Canada L8V 1C3.
authors.forename: P A
authors.initials: PA
authors.lastname: Outinen
authors.forename: S K
authors.initials: SK
authors.lastname: Sood
authors.forename: P C
authors.initials: PC
authors.lastname: Liaw
authors.forename: K D
authors.initials: KD
authors.lastname: Sarge
authors.forename: N
authors.initials: N
authors.lastname: Maeda
authors.forename: J
authors.initials: J
authors.lastname: Hirsh
authors.forename: J
authors.initials: J
authors.lastname: Ribau
authors.forename: T J
authors.initials: TJ
authors.lastname: Podor
authors.forename: J I
authors.initials: JI
authors.lastname: Weitz
authors.forename: R C
authors.initials: RC
authors.lastname: Austin
mesh_terms: D000818:Animals; D022762:CCAAT-Enhancer-Binding Proteins; D002352:Carrier Proteins / Q000235:genetics; D002478:Cells, Cultured; D003541:Cystathionine beta-Synthase / Q000172:deficiency; D004268:DNA-Binding Proteins / Q000235:genetics; D004195:Disease Models, Animal; D000091342:Endoplasmic Reticulum Chaperone BiP; D004730:Endothelium, Vascular / Q000187:drug effects*; D005786:Gene Expression Regulation / Q000187:drug effects*; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism; D006360:Heat-Shock Proteins; D006710:Homocysteine / Q000097:blood / Q000494:pharmacology*; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007150:Immunohistochemistry; D051379:Mice; D018832:Molecular Chaperones / Q000235:genetics; D009130:Muscle, Smooth / Q000187:drug effects; D012333:RNA, Messenger / Q000187:drug effects; D013312:Stress, Physiological / Q000378:metabolism*; D051742:Transcription Factor CHOP; D014157:Transcription Factors / Q000235:genetics
chemical_list: D022762:CCAAT-Enhancer-Binding Proteins; D002352:Carrier Proteins; C497249:DDIT3 protein, human; D004268:DNA-Binding Proteins; C497250:Ddit3 protein, mouse; D000091342:Endoplasmic Reticulum Chaperone BiP; D018840:HSP70 Heat-Shock Proteins; C000717247:HSPA5 protein, human; D006360:Heat-Shock Proteins; C000717248:Hspa5 protein, mouse; D018832:Molecular Chaperones; D012333:RNA, Messenger; D014157:Transcription Factors; D006710:Homocysteine; D051742:Transcription Factor CHOP; D006861:Hydrogen Peroxide; D003541:Cystathionine beta-Synthase