The role of SHP/REV-ERBα/CYP4A axis in the pathogenesis of alcohol-associated liver disease
Zhihong Yang, Rana V. Smalling, Yi Huang, Yanchao Jiang, Praveen Kusumanchi, Will Bogaert, Li Wang, Don A. Delker, Nicholas J. Skill, Sen Han, Ting Zhang, Jing Ma, Nazmul Huda, Suthat Liangpunsakul
Zhihong Yang, Rana V. Smalling, Yi Huang, Yanchao Jiang, Praveen Kusumanchi, Will Bogaert, Li Wang, Don A. Delker, Nicholas J. Skill, Sen Han, Ting Zhang, Jing Ma, Nazmul Huda, Suthat Liangpunsakul
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Research Article Hepatology

The role of SHP/REV-ERBα/CYP4A axis in the pathogenesis of alcohol-associated liver disease

Abstract

Alcohol-associated liver disease (ALD) represents a spectrum of histopathological changes, including alcoholic steatosis, steatohepatitis, and cirrhosis. One of the early responses to excessive alcohol consumption is lipid accumulation in the hepatocytes. Lipid ω-hydroxylation of medium- and long-chain fatty acid metabolized by the cytochrome P450 4A (CYP4A) family is an alternative pathway for fatty acid metabolism. The molecular mechanisms of CYP4A in ALD pathogenesis have not been elucidated. In this study, WT and Shp−/− mice were fed with a modified ethanol-binge, National Institute on Alcohol Abuse and Alcoholism model (10 days of ethanol feeding plus single binge). Liver tissues were collected every 6 hours for 24 hours and analyzed using RNA-Seq. The effects of REV-ERBα agonist (SR9009, 100 mg/kg/d) or CYP4A antagonist (HET0016, 5 mg/kg/d) in ethanol-fed mice were also evaluated. We found that hepatic Cyp4a10 and Cyp4a14 expression were significantly upregulated in WT mice, but not in Shp−/− mice, fed with ethanol. ChIP quantitative PCR and promoter assay revealed that REV-ERBα is the transcriptional repressor of Cyp4a10 and Cyp4a14. Rev-Erbα−/− hepatocytes had a marked induction of both Cyp4a genes and lipid accumulation. REV-ERBα agonist SR9009 or CYP4A antagonist HET0016 attenuated Cyp4a induction by ethanol and prevented alcohol-induced steatosis. Here, we have identified a role for the SHP/REV-ERBα/CYP4A axis in the pathogenesis of ALD. Our data also suggest REV-ERBα or CYP4A as the potential therapeutic targets for ALD.

Authors

Zhihong Yang, Rana V. Smalling, Yi Huang, Yanchao Jiang, Praveen Kusumanchi, Will Bogaert, Li Wang, Don A. Delker, Nicholas J. Skill, Sen Han, Ting Zhang, Jing Ma, Nazmul Huda, Suthat Liangpunsakul

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Figure 3

Deficiency of REV-ERBα largely induced both Cyp4a10 and Cyp4a14 expression, promoted lipid accumulation and oxidative stress.

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Deficiency of REV-ERBα largely induced both Cyp4a10 and Cyp4a14 expressi...
(A) qPCR analysis of hepatic Cyp4a10 and Cyp4a14 mRNAs in WT or Rev-Erbα−/− mice ***P < 0.001; ****P < 0.0001 versus WT. Two-tailed Student’s t test. (B) Western blot analysis of CYP4A protein from WT and Rev-Erbα−/− primary hepatocytes treated with or without 50 mM ethanol (Etoh) for 24 hours. (C) Oil Red O (left) and Nile Red (right) staining in primary hepatocytes of WT or Rev-Erbα−/− mice treated with or without 50 mM ethanol for 24 hours. Scale bar: 100 μm. (D) The quantification of the positive area (% to total area) from Nile Red staining (right). *P < 0.05, **P < 0.01 versus WT-E(-); ##P < 0.05 versus WT-E(+). One-way ANOVA. (E) ROS generation from WT and Rev-Erbα−/− primary hepatocytes treated with or without 50 mM Ethanol for indicated times. *P < 0.05, **P < 0.01 versus WT. One-way ANOVA.