Molecular mechanism of the short-term cardiotoxicity caused by 2',3'-dideoxycytidine (ddC): modulation of reactive oxygen species levels and ADP-ribosylation reactions.
The short-term cardiac side effects of 2',3'-dideoxycytidine (ddC, zalcitabine) were studied in rats in order to understand the biochemical events contributing to the development of ddC-induced cardiomyopathy. In developing animals, ddC treatment provoked a surprisingly rapid appearance of cardiac malfunctions characterized by prolonged RR, PR, and QT intervals and J point depression. The energy metabolism in the heart was compromised, characterized by a decreased creatine phosphate/creatine ratio (from 2.05 normal value to 0.75) and a decreased free ATP/ADP ratio (from 332 normal value to 121). The activity of respiratory complexes (NADH: cytochrome c oxidoreductase and cytochrome oxidase) also decreased significantly. Southern blot and polymerase chain reaction analysis did not show deletions or a decrease in the quantity of mitochondrial DNA (mtDNA) deriving from ddC-treated rat hearts, indicating that under our experimental conditions, ddC-induced heart abnormalities were not the direct consequence of mtDNA-related damage. The ddC treatment of rats significantly increased the formation of reactive oxygen species (ROS) in heart and skeletal muscle as determined by the oxidation of non-fluorescent dihydrorhodamine123 to fluorescent rhodamine123 and the oxidation of cellular proteins determined from protein carbonyl content. An activation of the nuclear poly-(ADP-ribose) polymerase (EC 2.4.2.30) and an increase in the mono-ADP-ribosylation of glucose-regulated protein and desmin were observed in the cardiac tissue from ddC-treated animals. A decrease in the quantity of heat shock protein (HSP)70s was also detected, while the level of HSP25 and HSP60 remained unchanged. Surprisingly, ddC treatment induced a skeletal muscle-specific decrease in the quantity of three proteins, one of which was identified by N-terminal sequencing as myoglobin, and another by tandem mass spectrometer sequencing as triosephosphate isomerase (EC 5.3.1.1). These data show that the short term cardiotoxicity of ddC is partially based on ROS-mediated signalling through poly- and mono-ADP-ribosylation reactions and depression of HSP70 levels, whose processes represent a new mtDNA independent mechanism for ddC-induced cell damage.
Skuta
G
G
Department of Biochemistry, University Medical School, Pecs, Hungary.
Fischer
G M
GM
Janaky
T
T
Kele
Z
Z
Szabo
P
P
Tozser
J
J
Sumegi
B
B
D000246:Adenosine Diphosphate Ribose / Q000378:metabolism*; D000818:Animals; D019380:Anti-HIV Agents / Q000633:toxicity*; D009202:Cardiomyopathies / Q000139:chemically induced* / Q000378:metabolism; D004247:DNA / Q000187:drug effects / Q000378:metabolism; D004562:Electrocardiography / Q000187:drug effects; D004734:Energy Metabolism / Q000187:drug effects; D006321:Heart / Q000187:drug effects* / Q000502:physiology; D006360:Heat-Shock Proteins / Q000187:drug effects / Q000378:metabolism; D013058:Mass Spectrometry; D008928:Mitochondria / Q000187:drug effects / Q000201:enzymology; D018482:Muscle, Skeletal / Q000187:drug effects / Q000378:metabolism; D010084:Oxidation-Reduction; D051381:Rats; D017208:Rats, Wistar; D017382:Reactive Oxygen Species / Q000378:metabolism*; D018894:Reverse Transcriptase Inhibitors / Q000633:toxicity; D017421:Sequence Analysis; D016047:Zalcitabine / Q000633:toxicity*
D019380:Anti-HIV Agents; D006360:Heat-Shock Proteins; D017382:Reactive Oxygen Species; D018894:Reverse Transcriptase Inhibitors; D000246:Adenosine Diphosphate Ribose; D016047:Zalcitabine; D004247:DNA