Elevated levels of oxidative DNA damage and DNA repair enzymes in human atherosclerotic plaques.
BACKGROUND
The formation of reactive oxygen species is a critical event in atherosclerosis because it promotes cell proliferation, hypertrophy, growth arrest, and/or apoptosis and oxidation of LDL. In the present study, we investigated whether reactive oxygen species-induced oxidative damage to DNA occurs in human atherosclerotic plaques and whether this is accompanied by the upregulation of DNA repair mechanisms.


RESULTS
We observed increased immunoreactivity against the oxidative DNA damage marker 7,8-dihydro-8-oxo-2'-deoxyguanosine (8-oxo-dG) in plaques of the carotid artery compared with the adjacent inner media and nonatherosclerotic mammary arteries. Strong 8-oxo-dG immunoreactivity was found in all cell types of the plaque including macrophages, smooth muscle cells, and endothelial cells. As shown by competitive ELISA, carotid plaques contained 160+/-29 8-oxo-dG residues/10(5) dG versus 3+/-1 8-oxo-dG residues/10(5) dG in mammary arteries. Single-cell gel electrophoresis showed elevated levels of DNA strand breaks in the plaque. The overall number of apoptotic nuclei was low (1% to 2%) and did not correlate with the amount of 8-oxo-dG immunoreactive cells (>90%). This suggests that initial damage to DNA occurs at a sublethal level. Several DNA repair systems that are involved in base excision repair (redox factor/AP endonuclease [Ref 1] and poly(ADP-ribose) polymerase 1 [PARP-1]) or nonspecific repair pathways (p53, DNA-dependent protein kinase) were upregulated, as shown by Western blotting and immunohistochemistry. Overexpression of DNA repair enzymes was associated with elevated levels of proliferating cell nuclear antigen.


CONCLUSIONS
Our findings provide evidence that oxidative DNA damage and repair increase significantly in human atherosclerotic plaques.
Division of Pharmacology, University of Antwerp, Wilrijk, Belgium.
Wim
W
Martinet
Michiel W M
MW
Knaapen
Guido R Y
GR
De Meyer
Arnold G
AG
Herman
Mark M
MM
Kockx
D000080242:8-Hydroxy-2'-Deoxyguanosine; D000368:Aged; D017209:Apoptosis; D001161:Arteriosclerosis / Q000201:enzymology / Q000235:genetics / Q000473:pathology*; D016893:Carotid Stenosis / Q000201:enzymology / Q000235:genetics / Q000473:pathology; D002467:Cell Nucleus / Q000737:chemistry / Q000648:ultrastructure; D004249:DNA Damage; D004260:DNA Repair; D003849:Deoxyguanosine / Q000031:analogs & derivatives / Q000032:analysis; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D018384:Oxidative Stress; D015854:Up-Regulation
D000080242:8-Hydroxy-2'-Deoxyguanosine; D003849:Deoxyguanosine