Epigenetic histone modifications in a clinically relevant rat model of chronic ethanol-binge-mediated liver injury
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Ethanol binge augments liver injury after chronic ethanol consumption in humans, but the mechanism behind the enhanced liver injury by ethanol binge is not known. In this study we used a clinically relevant rat model in which liver injury is amplified by binge after chronic ethanol treatment and investigated the importance of histone modifications.
Eight-week-old Sprague-Dawley rats were fed ethanol in a liquid diet for 4 weeks. Control rats were fed an isocaloric liquid diet. This was followed by three binge administrations of ethanol (intragastric 5 g/kg body weight, 12 h apart). In the control, ethanol was replaced by water. Four hours after the last binge administration, liver samples were analyzed for histone modifications and parameters of liver injury.
Chronic ethanol administration alone caused an increase in histone H3 ser10 and ser28 (H3S10 or S28) phosphorylation, and binge ethanol reduced their levels. Levels of dually modified phosphoacetylated histone H3 (H3AcK9/PS10) increased after acute binge ethanol and remained same after chronic ethanol binge. In contrast, histone H3 lysine-9 acetylation (H3AcK9) was not increased after chronic ethanol but increased significantly after acute binge and chronic ethanol binge. Increase in histone acetylation was accompanied by increased phospho-ERK1/2 in the nuclear extracts. Increased acetylation after chronic ethanol binge was also accompanied by increased protein levels of GCN5 histone acetyl transferase and a modest increase in HDAC3 in the nucleus. Histone lysine-9 dimethylation was significantly increased after chronic ethanol binge. Chronic ethanol binge also resulted in a decrease in the SAM:SAH ratio with a relative decrease of SAM levels and a corresponding increase in SAH levels.
Ethanol binge after chronic ethanol altered the profile of site-specific histone modifications and may underlie the mechanism of augmented liver injury by chronic-ethanol-binge-treated rats.
KeywordsAlcoholic steatohepatitis Binge ethanol Epigenetics Histone modifications Necrosis
This work was supported by NIH grant AA16347 (SDS) and in part by MERIT Review grant BX001155 (KKK) from the Department of Veterans Affairs, Office of Research and Development (Biomedical Laboratory Research & Development).
Compliance with ethical requirements and Conflict of interest
In this study the protocol for the use of rats was approved by the University of Missouri Animal Care and Use Committee (approval no. 7178). This committee follows the guidelines set by the National Institutes of Health (USA) for the use and care of laboratory animals. Annayya R. Aroor, Ricardo J. Restrepo, Kususm K. Kharbanda, and Shivendra D. Shukla declare that they have no conflict of interest.
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