Oxidative stress-mediated apoptosis of hepatocytes exposed to acute ethanol intoxication

Iwao Kurose, Hajime Higuchi, Soichiro Miura, Hidetsugu Saito, Naoyuki Watanabe, Ryota Hokari, Masahiko Hirokawa, Masaaki Takaishi, Sigeyuki Zeki, Tetsuya Nakamura, Hirotoshi Ebinuma, Shinzo Kato, Hiromasa Ishii

研究成果: Article査読

189 被引用数 (Scopus)

抄録

The present study was designed to investigate whether acute ethanol intoxication increases the production of active oxidants, and subsequently promotes apoptosis of hepatocytes. Hepatocytes were isolated from male Wistar rats, and cultured in the presence or absence of ethanol. The fluorescence in situ nick end labeling method and an enzyme-linked immunosorbent assay (ELISA) system to quantify fragmented DNA were used to estimate apoptotic change in hepatocytes. Nuclear morphological alterations and membrane barrier dysfunction of hepatocytes were assessed by staining with Hoechst 33342 and propidium iodide (PI). Intracellular glutathione level was determined as the fluorescence of monochlorobimane (MCLB), which forms conjugate with glutathione to become fluorescent. Ethanol (100 mmol/l) increased the amount of fragmented Dna and the number of apoptotic hepatocytes in vivo as well as in vitro. These ethanol-induced alterations in hepatocytes were attenuated by simultaneous incubation with either 4-methylpyrazole, an inhibitor of alcohol dehydrogenase, or dimethylthiourea, an intracellular oxidant scavenger. Diethyl maleic acid (DMA), a glutathione depictor, enhanced the induction of apoptotic change, and decreased membrane barrier function in ethanol-treated hepatocytes, whereas ethanol per se did not increase the number of PI- positive hepatocytes. Furthermore, combination of ethanol and DMA but not ethanol alone decreased the hepatocyte MCLB fluorescence. Taken together, the present study suggests that active oxidants produced during ethanol metabolism mediate fragmentation of DNA in hepatocytes, and that intracellular antioxidants such as glutathione play a critical role in the cytoprotective mechanisms of hepatocyte against lethal cell death, ie, apoptosis, induced by ethanol.

本文言語English
ページ(範囲)368-378
ページ数11
ジャーナルHepatology
25
2
DOI
出版ステータスPublished - 1997 2月
外部発表はい

ASJC Scopus subject areas

  • 肝臓学

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