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4-Hydroxynonenal and Malondialdehyde Hepatic Protein Adducts in Rats Treated with Carbon Tetrachloride: Immunochemical Detection and Lobular Localization

Authors
Journal
Toxicology and Applied Pharmacology
0041-008X
Publisher
Elsevier
Publication Date
Volume
161
Issue
1
Identifiers
DOI: 10.1006/taap.1999.8788
Keywords
  • Carbon Tetrachloride
  • Lipid Peroxidation
  • Hepatotoxicity
  • Malondialdehyde-4-Hydroxynonenal-Protein Adducts
  • Oxidative Stress
Disciplines
  • Biology
  • Chemistry

Abstract

Abstract The metabolism of CCl 4 initiates the peroxidation of polyunsaturated fatty acids producing α,β-unsaturated aldehydes, such as 4-hydroxynonenal (4-HNE) and malondialdehyde (MDA). The facile reactivity of these electrophilic aldehydic products suggests they play a role in the toxicity of compounds like CCl 4. To determine the rate at which CCl 4-initiated lipid peroxidation results in the formation of 4-HNE and/or MDA hepatic protein adducts, rats were given an intragastric dose of CCl 4 (1.0 ml/kg) and euthanized 0–72 h after administration. Rabbit polyclonal antisera directed toward 4-HNE- or MDA-protein epitopes were employed in immuno-histochemical and immuno-precipitation/Western analyses to detect 4-HNE and MDA-protein adducts in paraffin-embedded liver sections and liver homogenates. As early as 6 h post CCl 4 exposure, 4-HNE and MDA adducts were detected immuno-histochemically in hepatocytes localized to zone 2 of the hepatic acinus. Liver injury was progressive to 24 h as lipid peroxidation and hepatocellular necrosis increased. The hallmark of CCl 4 hepatotoxicity, zone 3 necrosis, was observed 24 h after CCl 4 administration and immuno-positive hepatocytes were observed in zone 2 as well as zone 3. Immuno-positive cells were no longer visible by 36 to 72 h post CCl 4 administration. From 6 to 48 h after CCl 4 administration, at least four adducted proteins were immuno-precipitated from liver homogenates with the anti-MDA or anti-4HNE serum, which corresponded to molecular weights of 80, 150, 205, and greater than 205 kDa. These results demonstrate that 4-HNE and MDA alkylate specific hepatic proteins in a time-dependent manner, which appears to be associated with hepatocellular injury following CCl 4 exposure.

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