Zonal distribution of protein-acetaldehyde adducts in the liver of rats fed alcohol for long periods

Renee C. Lin, Feng Zhou, Michael J. Fillenwarth, Lawrence Lumeng

Research output: Contribution to journalArticle

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Abstract

Acetaldehyde, a highly reactive intermediate of alcohol metabolism, has been shown to form adducts with liver proteins in rats fed alcohol for long periods. In this report, the zonal distribution of liver protein-acetaldehyde adducts that formed in vivo was studied by means of histoimmunostaining. Rats were pair-fed alcohol-containing and alcohol-free AIN'76 liquid diets for 2 or 11 wk before they were killed and subjected to whole body perfusion with paraformaldehyde. Each liver was cut into 60-μm-thick slices. Slices were first treated with 10% hydrogen peroxide to eliminate endogenous peroxidase activity. They were then incubated sequentially with rabbit antihemocyanin-acetaldehyde adduct, goat antirabbit serum IgG and rabbit peroxidase-antiperoxidase complex. The liver slices were stained with diaminobenzidine and counterstained with methylgreen. In the livers of rats fed alcohol for 2 wk, peroxidase activity was evident in the perivenous zone but not the periportal zone. No staining was obtained when the primary antibody had been preabsorbed with immobilized hemocyanin-acetaldehyde adduct or if the liver slices were incubated with the unimmunized rabbit IgG. Slight staining of the perivenous zone was seen in the livers of control rats, presumably because of minimal protein-acetaldehyde adduct formation emanating from endogenous acetaldehyde. When rats were fed alcohol for longer periods (e.g., 11 wk), protein-acetaldehyde adducts were still seen predominantly in the perivenous zone, but the distribution pattern was more diffuse than that observed in the livers of rats fed alcohol for only 2 wk. More liver cells produced protein-acetaldehyde adducts when rats were fed the alcohol-containing diet supplemented with cyanamide. However, these protein-acetaldehyde adduct-positive cells were still found mainly in the perivenous area. The zonation in the formation of protein-acetaldehyde adducts in the liver may in part explain the preferential damage of perivenous hepatocytes induced by long-term alcohol consumption.

Original languageEnglish
Pages (from-to)864-869
Number of pages6
JournalHepatology
Volume18
Issue number4
StatePublished - Oct 1993

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Acetaldehyde
Alcohols
Liver
Proteins
Peroxidase
Rabbits
Immunoglobulin G
Cyanamide
Staining and Labeling
Diet
Hemocyanin
Goats
Alcohol Drinking
Hydrogen Peroxide
Hepatocytes
Perfusion

ASJC Scopus subject areas

  • Hepatology

Cite this

Zonal distribution of protein-acetaldehyde adducts in the liver of rats fed alcohol for long periods. / Lin, Renee C.; Zhou, Feng; Fillenwarth, Michael J.; Lumeng, Lawrence.

In: Hepatology, Vol. 18, No. 4, 10.1993, p. 864-869.

Research output: Contribution to journalArticle

Lin, Renee C. ; Zhou, Feng ; Fillenwarth, Michael J. ; Lumeng, Lawrence. / Zonal distribution of protein-acetaldehyde adducts in the liver of rats fed alcohol for long periods. In: Hepatology. 1993 ; Vol. 18, No. 4. pp. 864-869.
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abstract = "Acetaldehyde, a highly reactive intermediate of alcohol metabolism, has been shown to form adducts with liver proteins in rats fed alcohol for long periods. In this report, the zonal distribution of liver protein-acetaldehyde adducts that formed in vivo was studied by means of histoimmunostaining. Rats were pair-fed alcohol-containing and alcohol-free AIN'76 liquid diets for 2 or 11 wk before they were killed and subjected to whole body perfusion with paraformaldehyde. Each liver was cut into 60-μm-thick slices. Slices were first treated with 10{\%} hydrogen peroxide to eliminate endogenous peroxidase activity. They were then incubated sequentially with rabbit antihemocyanin-acetaldehyde adduct, goat antirabbit serum IgG and rabbit peroxidase-antiperoxidase complex. The liver slices were stained with diaminobenzidine and counterstained with methylgreen. In the livers of rats fed alcohol for 2 wk, peroxidase activity was evident in the perivenous zone but not the periportal zone. No staining was obtained when the primary antibody had been preabsorbed with immobilized hemocyanin-acetaldehyde adduct or if the liver slices were incubated with the unimmunized rabbit IgG. Slight staining of the perivenous zone was seen in the livers of control rats, presumably because of minimal protein-acetaldehyde adduct formation emanating from endogenous acetaldehyde. When rats were fed alcohol for longer periods (e.g., 11 wk), protein-acetaldehyde adducts were still seen predominantly in the perivenous zone, but the distribution pattern was more diffuse than that observed in the livers of rats fed alcohol for only 2 wk. More liver cells produced protein-acetaldehyde adducts when rats were fed the alcohol-containing diet supplemented with cyanamide. However, these protein-acetaldehyde adduct-positive cells were still found mainly in the perivenous area. The zonation in the formation of protein-acetaldehyde adducts in the liver may in part explain the preferential damage of perivenous hepatocytes induced by long-term alcohol consumption.",
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