Mitochondrial function is involved in LDL oxidation mediated by human cultured endothelial cells.
Human endothelial cells (ECs) grown under standard conditions are able to generate a basal level of oxygen free radicals and induce progressive oxidation of LDLs. Inhibition of cell-mediated LDL oxidation by superoxide dismutase, EDTA, or desferrioxamine implicates a role for superoxide anion and/or transition metals in this process. The potential role of the mitochondrion was investigated by inducing mitochondrial deenergization by selective photosensitization or the addition of inhibitors of the mitochondrial respiratory chain. Mitochondria of human cultured ECs were selectively damaged by photosensitization of cells labeled with the mitochondrion-selective fluorescent dye 2-(4-dimethylaminostyryl)-1-methylpyridinium iodide under conditions that induced only low levels of toxicity during the time of the experiment. Photosensitized ECs exhibited severe mitochondrial dysfunction, as suggested by the defect in mitochondrial uptake of the mitochondrion-selective fluorescent dyes [rhodamine 123 and 2-(4-dimethylaminostyryl)-1-methylpyridinium iodide] and morphological alterations as shown by transmission electron microscopy. In mitochondria-photosensitized cells, superoxide anion generation was strongly decreased, as was LDL oxidation and the subsequent cytotoxicity. When ECs were incubated with the mitochondrial respiratory-chain inhibitors antimycin A or rotenone or with the carbonylcyanide-m-chlorophenylhydrazone uncoupler rhodamine 123, uptake and subcellular distribution were altered, and concomitantly superoxide anion production and LDL oxidation were strongly decreased. In conclusion, these data suggest that mitochondrial function is required, directly or indirectly, for the production of superoxide anion and the subsequent LDL oxidation by human vascular ECs.
Mabile
L
L
Biochemistry Laboratory (INSERM CJF-9206), Institut Louis Bugnard, CHU Rangueil, Toulouse, France.
Meilhac
O
O
Escargueil-Blanc
I
I
Troly
M
M
Pieraggi
M T
MT
Salvayre
R
R
Nègre-Salvayre
A
A
D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone / Q000494:pharmacology; D002460:Cell Line; D008078:Cholesterol, LDL / Q000378:metabolism*; D003676:Deferoxamine / Q000494:pharmacology; D004727:Endothelium / Q000166:cytology* / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005456:Fluorescent Dyes / Q000494:pharmacology; D006801:Humans; D007476:Ionophores / Q000494:pharmacology; D008027:Light; D008353:Mannitol / Q000494:pharmacology; D008928:Mitochondria / Q000502:physiology* / Q000528:radiation effects; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D010084:Oxidation-Reduction; D013482:Superoxide Dismutase / Q000037:antagonists & inhibitors; D013481:Superoxides / Q000378:metabolism; D019323:omega-N-Methylarginine / Q000494:pharmacology
D008078:Cholesterol, LDL; D004791:Enzyme Inhibitors; D005456:Fluorescent Dyes; D007476:Ionophores; D013481:Superoxides; D019323:omega-N-Methylarginine; D008353:Mannitol; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone; D013482:Superoxide Dismutase; D003676:Deferoxamine; D019331:NG-Nitroarginine Methyl Ester