Increased reactive oxygen species production with antisense oligonucleotides directed against uncoupling protein 2 in murine endothelial cells.
Uncoupling protein 2 (UCP-2) belongs to the mitochondrial anion carrier family. It is ubiquitously expressed but is most abdundant in the reticuloendothelial system. In addition to uncoupling function, UCP-2 modulates the production of reactive oxygen species (ROS) by isolated mitochondria. Using an antisense oligonucleotide strategy, we investigated whether a defect in UCP-2 expression modulates ROS in intact endothelial cells. Murine endothelial cells (CRL 2181) pretreated by antisense oligonucleotides directed against UCP-2 mRNA exhibited a significant and specific increase in membrane potential and intracellular ROS level compared with control scrambled or anti-UCP-1 and -UCP-3 antisense oligonucleotides. These specific changes induced by UCP-2 antisense oligonucleotides were correlated with a rise in extracellular superoxide anion production and oxidative stress assessed by thiobarbituric acid reactive substance values. Taken together, these data suggest a role for UCP-2 in control of ROS production and subsequent oxidation of surrounding compounds mediating oxidative stress of endothelial cells. These data also support the notion that manipulations of UCP-2 at the genetic level could control ROS metabolism at the cellular level.
Duval
Carine
C
Unité Mixte recherche (UMR)-INSERM U466, Institut federatif de recherche (IFR) 31, Toulouse, France.
Nègre-Salvayre
Anne
A
Dogilo
Alain
A
Salvayre
Robert
R
Pénicaud
Luc
L
Casteilla
Louis
L
D000818:Animals; D002460:Cell Line; D015536:Down-Regulation; D004727:Endothelium / Q000166:cytology / Q000378:metabolism*; D007473:Ion Channels; D026901:Membrane Transport Proteins; D051379:Mice; D024101:Mitochondrial Proteins; D016376:Oligonucleotides, Antisense / Q000235:genetics / Q000378:metabolism*; D010084:Oxidation-Reduction; D011506:Proteins / Q000235:genetics* / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism*; D020112:Rhodamine 123; D017392:Thiobarbituric Acid Reactive Substances / Q000032:analysis; D000071246:Uncoupling Protein 2
D007473:Ion Channels; D026901:Membrane Transport Proteins; D024101:Mitochondrial Proteins; D016376:Oligonucleotides, Antisense; D011506:Proteins; D012333:RNA, Messenger; D017382:Reactive Oxygen Species; D017392:Thiobarbituric Acid Reactive Substances; C000604390:Ucp2 protein, mouse; D000071246:Uncoupling Protein 2; D020112:Rhodamine 123