Clinical concentrations of doxorubicin inhibit activity of myocardial membrane-associated, calcium-independent phospholipase A(2).
Use of the anticancer antibiotic doxorubicin continues to be limited by its cumulative dose-related cardiotoxicity. Our study reports inhibition of myocardial intracellular calcium-independent phospholipase A(2) (iPLA(2)) activity by clinically relevant concentrations of the drug. The effect was first shown in vitro using suspensions of freshly isolated rat and rabbit cardiomyocytes. Addition of 0.1-10 microM doxorubicin to these cells led to a concentration- and time-dependent inhibition of total iPLA(2), as measured using (16:0, [(3)H]18:1) plasmenylcholine and phosphatidylcholine substrates in the presence or absence of calcium. Subcellular fractionation into cytosolic and membrane fraction revealed that the drug selectively inhibits membrane-associated iPLA(2) activity, without altering activity of the cytosolic enzyme. Doxorubicin treatment of cells prelabeled with [H(3)]arachidonic acid led to a depression of baseline arachidonic acid release levels, corroborating iPLA(2) inhibition. Reducing agents blocked PLA(2) inhibition in cardiomyocyte suspensions, suggesting that the doxorubicin effect is mediated by oxidation of susceptible cysteines. In vivo experiments, in which adults rats were i.v. injected with a bolus dose of 4 mg/kg doxorubicin, confirmed in vitro findings, revealing a 2-fold decrease in membrane-associated Ca(2+)-independent iPLA(2) activity in the heart tissue of treated animals. The observed phenomenon has important implications for myocyte signaling cascades and membrane remodeling.
McHowat
J
J
Department of Pathology, St. Louis University School of Medicine, St. Louis, MO 63104, USA.
Swift
L M
LM
Arutunyan
A
A
Sarvazyan
N
N
D000818:Animals; D000903:Antibiotics, Antineoplastic / Q000633:toxicity*; D016718:Arachidonic Acid / Q000378:metabolism; D015153:Blotting, Western; D002462:Cell Membrane / Q000201:enzymology; D003600:Cytosol / Q000201:enzymology; D004229:Dithiothreitol / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004317:Doxorubicin / Q000633:toxicity*; D004357:Drug Synergism; D005978:Glutathione / Q000494:pharmacology; D054522:Group VI Phospholipases A2; D006321:Heart / Q000187:drug effects* / Q000378:metabolism; D006352:Heart Ventricles / Q000166:cytology / Q000187:drug effects / Q000201:enzymology; D008297:Male; D009206:Myocardium / Q000166:cytology / Q000201:enzymology*; D009281:Naphthalenes / Q000633:toxicity; D010726:Phosphodiesterase Inhibitors / Q000633:toxicity; D010741:Phospholipases A / Q000037:antagonists & inhibitors*; D011753:Pyrones / Q000633:toxicity; D051381:Rats; D017207:Rats, Sprague-Dawley
D000903:Antibiotics, Antineoplastic; D009281:Naphthalenes; D010726:Phosphodiesterase Inhibitors; D011753:Pyrones; D016718:Arachidonic Acid; D004317:Doxorubicin; C068314:6-(bromomethylene)tetrahydro-3-(1-naphthaleneyl)-2H-pyran-2-one; D010741:Phospholipases A; D054522:Group VI Phospholipases A2; D005978:Glutathione; D004229:Dithiothreitol