Acetaminophen-induced cytotoxicity in cultured mouse hepatocytes: correlation of nuclear Ca2+ accumulation and early DNA fragmentation with cell death.
Acetaminophen-induced cytotoxicity in cultured mouse hepatocytes: correlation of nuclear Ca2+ accumulation and early DNA fragmentation with cell death.
复制标题
对乙酰氨基酚诱导的培养小鼠肝细胞的细胞毒性：核 Ca2 积累和早期 DNA 断裂与细胞死亡的相关性。
DOI：
10.1016/0041-008x(91)90028-d
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发表时间：
1991
影响因子：
3.8
通讯作者：
Corcoran,GB
中科院分区：
文献类型：
--
作者：
Shen,W;Kamendulis,LM;Ray,SD;Corcoran,GB
Hepatotoxic doses of acetaminophen cause widespread alkylation of liver and early loss of cytosolic Ca2+regulation. Although the precise location and target of lethal alkylation are not known, Ca2+accumulation is viewed as a possible link between cell alkylation and cell death. We have recently shown that Ca2+accumulates in the nucleus and that DNA fragments in vivo before the development of acetaminophen-induced necrosis in mice. The present study examined cultured hepatocytes for nuclear damage and its association with cell death in vitro. Positive results would argue for two key points. (1) Nonparenchymal cell damage does not explain DNA fragmentation induced by acetaminophen in vivo. (2) A chemical that causes necrosis can produce DNA damage considered characteristic of apoptosis. Hepatocytes from NIH Swiss mice were isolated by collagenase perfusion, cultured in Williams' E medium for 24 hr, and exposed to acetaminophen. Cytotoxicity was assessed by lactate dehydrogenase leakage and release of [3H]adenine from a prelabeled nucleotide pool. Genomic DNA fragmentation was assessed quantitatively by colorimetric analysis and qualitatively by agarose gel electrophoresis. Acetaminophen caused DNA damage from 1–4 hr onward and produced significant release of lactate dehydrogenase and [3H]adenine nucleotides at later times. Agarose gel electrophoresis revealed a “ladder” of DNA fragments characteristic of Ca2+-mediated endonuclease activation. Cytotoxicity correlated with nuclear Ca2+accumulation (r > 0.895, p < 0.05) and with percentage DNA fragmentation (r > 0.835, p < 0.05). Nuclear changes in vitro generally reproduced those observed in vivo. Collectively, these findings demonstrate that nuclear Ca2+accumulation and DNA fragmentation appear as early events that correlate directly with later cytotoxicity. These changes may contribute to acetaminophen-induced injury leading to cell death in vitro and necrosis in vivo.
