Abstract
The epigenetic histone modification by ethanol is emerging as one of the mechanisms for its deleterious effects in the liver. In this context, we have investigated the role of histone H3 phosphorylation at Ser10 (P-H3-Ser10), and Ser28 (P-H3-Ser28) in liver after acute ethanol treatment in vivo. Ethanol was administered intraperitoneally in male Sprague-Dawley rats. Ethanol dose-response (1–5 g/kg body weight) and time-course (1–4 h) experiments were conducted, and various parameters were monitored. Steatosis and necrosis (serum alanine aminotransferase) of the liver increased in 4 h, suggesting liver injury. There were differences between P-H3-Ser10 and P-H3-Ser28 at 1 h, with the latter being more sensitive to lower ethanol doses. It was noteworthy that phosphorylation of both serines disappeared at the highest dose used (5 g/kg). We also examined phosphoacetylation of histone H3 at K9S10 and observed a dramatic increase. The changes in histone H3 phosphorylation and phosphoacetylation were also accompanied with expression of early response genes (c-fos, c-jun, mitogen-activated protein kinase phosphatase-1). Chromatin immunoprecipitation assays in samples from 1.5 and 4 h of ethanol administration indicated that increased histone H3 phosphorylation at Ser28 was associated with the promoters of c-jun and plasminogen activator inhibitor-1. In conclusion, this study demonstrates for the first time that in vivo exposure of liver to acute ethanol induced phosphorylation and phosphoacetylation of histone H3, and these modifications are differentially involved in the mRNA expression of genes.
Footnotes
This work was supported in part by the National Institutes of Health National Institute of Alcohol Abuse and Alcoholism [Grant AA16347].
Article, publication date, and citation information can be found at http://jpet.aspetjournals.org.
-
ABBREVIATIONS:
- MAPK
- mitogen-activated protein kinase
- ALT
- alanine aminotransferase
- AP-1
- activator protein-1
- BAC
- blood alcohol concentration
- bp
- base pairs
- ChIP
- chromatin immunoprecipitation
- ERK1/2
- extracellular signal-regulated kinase 1/2
- GAPDH
- glyceraldehyde-3-phosphate dehydrogenase
- HAT
- histone acetyltransferase
- JNK
- c-jun NH2-terminal kinase
- LDL-r
- low-density lipoprotein receptor
- MKP-1
- mitogen-activated protein kinase phosphatase-1
- NaF
- sodium fluoride
- PAGE
- polyacrylamide gel electrophoresis
- PAI-1
- plasminogen activator inhibitor-1
- PBS
- phosphate-buffered saline
- qRT-PCR
- quantitative real-time polymerase chain reaction
- TBST
- 20 mM Tris, pH 7.4, containing 0.1% Tween 20 and 150 mM NaCl
- TNF
- tumor necrosis factor
- TSA
- trichostatin A.
- Received August 8, 2011.
- Accepted October 21, 2011.
- Copyright © 2012 by The American Society for Pharmacology and Experimental Therapeutics
JPET articles become freely available 12 months after publication, and remain freely available for 5 years.Non-open access articles that fall outside this five year window are available only to institutional subscribers and current ASPET members, or through the article purchase feature at the bottom of the page.
|