Cocaine induces apoptosis in human coronary artery endothelial cells.
This study was designed to determine the direct cytotoxic effect of cocaine on human coronary artery endothelial cells (HCAECs). Cocaine treatment of cultured HCAECs induced a time- and dose-dependent increase in apoptotic cell death in HCAECs. Cocaine-induced surface exposure of phosphatidylserine in HCAECs was seen as early as at 6 h. With prolonged treatment < or =72 h, cocaine (10-500 microM) produced a dose-dependent increase in apoptosis in the cells. Corresponding DNA fragmentation induced by cocaine was demonstrated in situ by terminal deoxynucleotidyl transferase (Tdt) UTP nick end-labeling TUNEL assay and by electrophoresis of labeled DNA fragments, showing the characteristic apoptotic ladders. Both caspase-9 (Z-LEHD-FMK) and caspase-3 (Ac-DEVD-CHO) inhibitors blocked cocaine-induced apoptosis. In addition, cyclosporin A inhibited cocaine-induced apoptosis in a concentration-dependent manner with a median inhibitory concentration (IC50) of 0.3 microM. The maximum of 62% inhibition was obtained with 3 microM cyclosporin A. Cocaine-induced apoptosis also was blocked by naloxone and nifedipine in a dose-dependent manner. These findings suggest that cocaine induces apoptosis in cultured HCAECs, which may be mediated by opioid receptors. The release of cytochrome c from the mitochondria and its subsequent activation of caspase-9 and caspase-3 may play a key role in cocaine-induced apoptosis.
He
J
J
Department of Pharmacology, Loma Linda University School of Medicine, California 92350, USA.
Xiao
Y
Y
Zhang
L
L
D017209:Apoptosis; D061945:Caspase Inhibitors; D002478:Cells, Cultured; D003042:Cocaine / Q000494:pharmacology*; D003331:Coronary Vessels / Q000166:cytology; D016572:Cyclosporine / Q000494:pharmacology; D004347:Drug Interactions; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006801:Humans; D009270:Naloxone / Q000494:pharmacology; D009292:Narcotic Antagonists / Q000494:pharmacology; D009543:Nifedipine / Q000494:pharmacology; D014662:Vasoconstrictor Agents / Q000494:pharmacology*; D014665:Vasodilator Agents / Q000494:pharmacology
D061945:Caspase Inhibitors; D004791:Enzyme Inhibitors; D009292:Narcotic Antagonists; D014662:Vasoconstrictor Agents; D014665:Vasodilator Agents; D009270:Naloxone; D016572:Cyclosporine; D003042:Cocaine; D009543:Nifedipine