Plasma membrane hydroxyethyl radical adducts cause antibody-dependent cytotoxicity in rat hepatocytes exposed to alcohol
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Background & aims We reported previously that patients with alcoholic liver disease (ALD) have circulating immunoglobulins reacting with cytochrome P4502E1 (CYP2E1) complexed with hydroxyethyl free radicals. The aim of this study was to investigate whether hydroxyethyl radical adducts are present on the plasma membranes of ethanol-treated hepatocytes and their role in antibody-dependent cytotoxicity.Methods Immunofluorescence confocal laser microscopy, Western blotting, and antibody-dependent cell-mediated cytotoxicity assay were used.Results Isolated rat hepatocytes incubated in vitro with ethanol or obtained from ethanol-treated animals showed strong surface fluorescence when exposed to rabbit anti-hydroxyethyl radical serum or sera from patients with ALD. No surface fluorescence was evident on control hepatocytes or after scavenging hydroxyethyl radicals with 4-pyridyl-1-oxide-t-butyl nitrone. The presence of CYP2E1-hydroxyethyl radical adducts on hepatocyte plasma membranes was shown by Western blot and by immunofluorescence using double staining for human and rabbit anti-CYP2E1 immunoglobulin G. Cytotoxicity was observed in ethanol-treated hepatocytes incubated with immunoglobulin G from patients with ALD and normal human blood mononuclear cells. This effect was blocked by preabsorbing the sera with human albumin complexed with hydroxyethyl radicals, which also eliminated the antibody reaction with the plasma membranes.Conclusions Hydroxyethyl radicals bound to CYP2E1 on hepatocyte plasma membranes can target immune reactions triggered by alcohol abuse.
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Adducts derived from HER are detected in the liver and localized in microsomes and plasma membranes of hepatocytes and are inducible by alcohol exposure.
“Finally, adducts derived from HER are also detected in the liver and localized in the microsomes and plasma membranes of hepatocytes and are inducible by alcohol exposure229,230.”
From Advanced glycation end products (AGEs) and other adducts in aging-related diseases and alcohol-mediated tissue injury (2021) · cites this paper as [230]
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- Authors
- Clot, P., Parola, M., Bellomo, G., Dianzani, U., Carini, R., Tabone, M., Arico, S., Ingelman- Sundberg, M., & Albano, E.
- Affiliations
- University of Turin (IT)
- Venue
- Gastroenterology
- Publisher
- Elsevier BV
- Published
- 1997-07
- Volume
- 113
- Issue
- 1
- Pages
- 265-276
- ISSN
- 0016-5085 (ISSN-L)
- OpenAlex
- W2068601876
- Cited by
- 86
- References
- 4
- Citations by year
- 2022: 1 · 2021: 1 · 2018: 3 · 2016: 1 · 2015: 3 · 2014: 2 · 2013: 3 · 2012: 1
- FWCI
- 5.10
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Gastroenterology (publishedVersion)
- PubMed (publishedVersion)
- Institutional Research Information System University of Turin (University of Turin) (submittedVersion)
- Topics
- Health Sciences › Medicine › Pathology and Forensic Medicine › Alcohol Consumption and Health Effects
- Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Pharmacology › Drug-Induced Hepatotoxicity and Protection
- Health Sciences › Medicine › Hepatology › Liver physiology and pathology
- Keywords
- CYP2E1, alcoholic liver disease, antibody-dependent cellular cytotoxicity, hepatocyte plasma membrane, ethanol, autoantibodies, alcohol abuse, IgG
- MeSH terms
- Ethanol, Animals, Antibody-Dependent Cell Cytotoxicity, Cell Membrane, Free Radicals, Humans, Liver, Liver Diseases, Alcoholic, Male, Microsomes, Liver, Rabbits, Blotting, Western, Rats, Sprague-Dawley, Microscopy, Confocal, Cytochrome P-450 CYP2E1, Rats
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