INTRACELLULAR CALCIUM DISRUPTION AS A SECONDARY EVENT IN ACETAMINOPHEN-INDUCED HEPATOTOXICITY

被引:28
作者
GREWAL, KK [1 ]
RACZ, WJ [1 ]
机构
[1] QUEENS UNIV, FAC MED, DEPT PHARMACOL & TOXICOL, KINGSTON K7L 3N6, ONTARIO, CANADA
关键词
ACETAMINOPHEN; HEPATOTOXICITY; N-ACETYLCYSTEINE; DITHIOTHREITOL; CALCIUM ION;
D O I
10.1139/y93-004
中图分类号
R9 [药学];
学科分类号
1007 ;
摘要
To examine a role for disturbances in intracellular calcium homeostasis in acetaminophen-induced hepatotoxicity, freshly isolated mouse hepatocytes were incubated with 1.0 mM acetaminophen for 1.5 h to allow for covalent binding and initiation of cell damage. The hepatocytes were then washed and the cells incubated in fresh medium containing either 2.0 mM N-acetylcysteine or 1.5 mM dithiothreitol for the duration of a 4-h incubation period. These agents were used as tools in the elucidation of the biochemical events responsible for acetaminophen-induced cell necrosis. The reduced protein sulfhydryl content, cytosolic [Ca2+], and plasma membrane integrity were quantitated. Acetaminophen produced protein sulfhydryl depletion, an increased cytosolic [Ca2+], and cell injury; however, cytotoxicity preceded the increase in [Ca2+]. Both N-acetylcysteine and dithiothreitol restored the acetaminophen-induced protein sulfhydryl loss. Dithiothreitol prevented both further cell injury and an increase in the cytosolic [Ca2+]. However, cell death and a subsequent increase in cytosolic [Ca2+] proceeded unabated following N-acetylcysteine addition. Although both agents restored protein sulfhydryl content, in view of their contrasting ultimate effects on cell viability the role of reduced protein sulfhydryl depletion in acetaminophen-induced hepatic injury requires further investigation. The increase in cytosolic [Ca2+] with acetaminophen alone and with subsequent N-acetylcysteine addition was determined to be a secondary event in cell injury because cytotoxicity occurred by 1.5 h; however, the increase in cytosolic [Ca2+] was not observed until, 2.5 h. Additional evidence for changes in cytosolic [Ca2+] as a secondary event was obtained by incubating the hepatocytes with acetaminophen in the presence of fura 2. Measurement of the fluorescent signal indicated that fura 2 effectively chelated the cytosolic Ca2+ yet did not prevent cell toxicity.
引用
收藏
页码:26 / 33
页数:8
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