title: Homocysteine-dependent alterations in mitochondrial gene expression, function and structure. Homocysteine and H2O2 act synergistically to enhance mitochondrial damage.
abstract: Mitochondrial abnormalities have been identified in hepatocytes of patients with hyperhomocysteinemia and in endothelial cells from the aortas of rats with diet-induced hyperhomocysteinemia. However, the mechanism by which homocysteine affects mitochondria is unknown. In this report, homocysteine-induced expression of the mitochondrial electron transport chain gene, cytochrome c oxidase III/ATPase 6,8 (CO3/ATPase 6,8), was identified in a human megakaryocytic cell line DAMI using mRNA differential display. Steady-state mRNA levels of CO3/ATPase 6,8, as well as other mitochondrial transcripts, were increased in DAMI cells by homocysteine in a concentration- and time-dependent manner. Despite an increase in mitochondrial RNA levels and changes in mitochondrial ultrastructure, no effect on either cell growth or mitochondrial respiration rates was observed in DAMI cells exposed to homocysteine at concentrations up to 1 mM. In contrast, 1 mM homocysteine in the presence of Cu2+, which is known to generate H2O2, significantly decreased mitochondrial RNA levels, caused gross morphological changes in mitochondrial ultrastructure, and inhibited both cell growth and mitochondrial respiration rates. However, precursors of cellular glutathione and preexposure to heat shock blocked the decrease in mitochondrial RNA levels caused by homocysteine and Cu2+. The observations that (i) homocysteine and H2O2, but not H2O2 alone, caused a decrease in mitochondrial RNA levels, (ii) intracellular levels of H2O2 were significantly increased in the presence of homocysteine and Cu2+, and (iii) catalase, but not free radical scavengers, prevented a decrease in mitochondrial RNA levels, provide evidence that homocysteine and H2O2 act synergistically to cause mitochondrial damage. Furthermore, our findings suggest that intracellular glutathione and heat shock proteins play a role in protecting mitochondria against the adverse effects elicited by homocysteine and H2O2.
authors.affiliation: Hamilton Civic Hospitals Research Centre and McMaster University, Hamilton, Ontario L8V 1C3, Canada. raustin@thrombosis.hhscr.org
authors.forename: R C
authors.initials: RC
authors.lastname: Austin
authors.forename: S K
authors.initials: SK
authors.lastname: Sood
authors.forename: A M
authors.initials: AM
authors.lastname: Dorward
authors.forename: G
authors.initials: G
authors.lastname: Singh
authors.forename: S G
authors.initials: SG
authors.lastname: Shaughnessy
authors.forename: S
authors.initials: S
authors.lastname: Pamidi
authors.forename: P A
authors.initials: PA
authors.lastname: Outinen
authors.forename: J I
authors.initials: JI
authors.lastname: Weitz
mesh_terms: D000818:Animals; D015152:Blotting, Northern; D002460:Cell Line; D002470:Cell Survival; D018834:Chaperonin 60 / Q000235:genetics / Q000378:metabolism; D003300:Copper / Q000494:pharmacology; D004272:DNA, Mitochondrial / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D004357:Drug Synergism; D005609:Free Radicals; D015870:Gene Expression / Q000187:drug effects; D006710:Homocysteine / Q000494:pharmacology*; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D008928:Mitochondria / Q000187:drug effects*; D010101:Oxygen Consumption; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D013329:Structure-Activity Relationship
chemical_list: D018834:Chaperonin 60; D004272:DNA, Mitochondrial; D005609:Free Radicals; D012333:RNA, Messenger; D006710:Homocysteine; D003300:Copper; D006861:Hydrogen Peroxide