Effects of nitric oxide donors on cybrids harbouring the mitochondrial myopathy, encephalopathy, lactic acidosis and stroke-like episodes (MELAS) A3243G mitochondrial DNA mutation.

Abstract

Reactive nitrogen and oxygen species (O2*-, H2O2, NO* and ONOO-) have been strongly implicated in the pathophysiology of neurodegenerative and mitochondrial diseases. In the present study, we examined the effects of nitrosative and/or nitrative stress generated by DETA-NO {(Z)-1-[2-aminoethyl-N-(2-ammonioethyl)amino]diazen-1-ium-1,2-diolate}, SIN-1 (3-morpholinosydnonimine hydrochloride) and SNP (sodium nitroprusside) on U87MG glioblastoma cybrids carrying wt (wild-type) and mutant [A3243G (Ala3243-->Gly)] mtDNA (mitochondrial genome) from a patient suffering from MELAS (mitochondrial myopathy, encephalopathy, lactic acidosis and stroke-like episodes). The mutant cybrids had reduced activity of cytochrome c oxidase, significantly lower ATP level and decreased mitochondrial membrane potential. However, endogenous levels of reactive oxygen species were very similar in all cybrids regardless of whether they carried the mtDNA defects or not. Furthermore, the cybrids were insensitive to the nitrosative and/or nitrative stress produced by either DETA-NO or SIN-1 alone. Cytotoxicity, however, was observed in response to SNP treatment and a combination of SIN-1 and glucose-deprivation. The mutant cybrids were significantly more sensitive to these insults compared with the wt controls. Ultrastructural examination of dying cells revealed several characteristic features of autophagic cell death. We concluded that nitrosative and/or nitrative stress alone were insufficient to trigger cytotoxicity in these cells, but cell death was observed with a combination of metabolic and nitrative stress. The vulnerability of the cybrids to these types of injury correlated with the cellular energy status, which were compromised by the MELAS mutation.

• Authors: Jagdeep K Sandhu, Caroline Sodja, Kevan McRae, Yan Li, Peter Rippstein, Yau-Huei Wei, Boleslaw Lach, Fay Lee, Septimiu Bucurescu, Mary-Ellen Harper, Marianna Sikorska
• Journal: The Biochemical journal (Biochem J) Vol. 391 Issue Pt 2 Pg. 191-202 (Oct 15 2005) ISSN: 1470-8728 [Electronic] England
• PMID: 15969653 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• 1-hydroxy-2-oxo-3,3-bis(2-aminoethyl)-1-triazene
• DNA, Mitochondrial
• Nitric Oxide Donors
• Reactive Nitrogen Species
• Reactive Oxygen Species
• Triazenes
• Nitroprusside
• linsidomine
• Adenosine Triphosphate
• Molsidomine

Topics

• Adenosine Triphosphate (metabolism)
• Cell Death
• Cell Survival
• DNA, Mitochondrial (genetics)
• Humans
• Hybrid Cells (drug effects, enzymology, metabolism, ultrastructure)
• MELAS Syndrome (genetics)
• Molsidomine (analogs & derivatives, pharmacology)
• Mutation
• Nitric Oxide Donors (pharmacology)
• Nitroprusside (pharmacology)
• Reactive Nitrogen Species (metabolism)
• Reactive Oxygen Species (metabolism)
• Triazenes (pharmacology)