Hepatic steatosis, or fatty liver, is strongly associated with metabolic syndrome. Understanding the mechanism of hepatic steatosis will help to prevent and treat this common medical problem. The aryl hydrocarbon receptor (AhR), highly expressed in the liver, is a transcriptional factor originally cloned as a "xenobiotic receptor." AhR regulates the expression of xenobiotic enzymes by binding to the dioxin response elements (DREs) present in target gene promoters. Subsequent studies suggest that AhR may also have endobiotic functions by affecting physiology, but the molecular mechanism for the endobiotic function of AhR remains largely unknown. Our preliminary results showed that: (1) Transgenic mice expressing the constitutively activated AhR (CA-AhR) exhibited hepatic steatosis when maintained in chow diet, a phenotype that has been recapitulated in wild type mice treated with the AhR agonist;(2) Activation of AhR in transgenic mice induced the expression of CD36/FAT, a fatty acid transporter important for hepatic fatty acid uptake and steatosis;(3) Activation of CD36 gene expression was also seen in wild type mice treated with the AhR agonist and this activation was abolished in AhR-/- mice;(4) Treatment of human hepatoma cells with AhR agonist induced the expression of CD36 and increased the uptake of free fatty acids;(5) The mouse and human CD36 gene promoters were activated by AhR;(6) Activation of AhR inhibited very-low density lipoprotein (VLDL)-triglyceride secretion;(7) Activation of AhR suppressed peroxisomal fatty acid 2-oxidation;and 8) CA-AhR transgenic mice in third generation of backcross to C57BL/6J showed spontaneous steatosis and signs of compromised glucose tolerance. Based on our preliminary data, we hypothesize that activation of AhR promotes hepatic steatosis through multiple mechanisms, including the activation of fatty acid transporter CD36, suppression of fatty acid oxidation, and inhibition of export of triglycerides. By using the "gain-of-function" CA-AhR transgenic, "loss-of-function" AhR-/- and CD36-/-, and AhR ligand-treated wild type mice, we propose four specific aims: (1) To determine whether activation of AhR is sufficient and necessary to induce hepatic steatosis;(2) To characterize AhR-induced hepatic steatosis;(3) To determine whether the fatty acid transporter CD36 is necessary for the steatotic effect of AhR;and (4) To determine the molecular mechanism by which AhR regulates the expression of CD36. To our knowledge, the current study represents the first attempt to determine the pathophysiological role of AhR in hepatic steatosis and associated metabolic abnormalities. The tetracycline inducible AhR transgenic mice, exhibiting fatty liver even when maintained in chow diet, represent a novel, convenient and reversible model of nonalcoholic fatty liver disease (NAFLD). It is hoped that results from this study may help to establish AhR and its target fatty acid transporter as novel therapeutic targets for fatty liver in human patients.

Public Health Relevance

The aryl hydrocarbon receptor (AhR) is a transcriptional factor originally cloned as a "xenobiotic receptor" to sense xenobiotic toxicants. The goal of this study is to determine whether AhR has an endobiotic role in promoting hepatic steatosis. Hepatic steatosis, or fatty liver, is a common medical problem strongly associated with metabolic syndrome. It is hoped that results from this study may help to establish AhR as a novel therapeutic target for fatty liver in human patients.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK083952-03
Application #
8248808
Study Section
Hepatobiliary Pathophysiology Study Section (HBPP)
Program Officer
Doo, Edward
Project Start
2010-05-01
Project End
2015-03-31
Budget Start
2012-04-01
Budget End
2013-03-31
Support Year
3
Fiscal Year
2012
Total Cost
$311,219
Indirect Cost
$105,794
Name
University of Pittsburgh
Department
Pharmacology
Type
Schools of Pharmacy
DUNS #
004514360
City
Pittsburgh
State
PA
Country
United States
Zip Code
15213
Elcombe, Clifford R; Peffer, Richard C; Wolf, Douglas C et al. (2014) Mode of action and human relevance analysis for nuclear receptor-mediated liver toxicity: A case study with phenobarbital as a model constitutive androstane receptor (CAR) activator. Crit Rev Toxicol 44:64-82
Ou, Zhimin; Jiang, Mengxi; Hu, Bingfang et al. (2014) Transcriptional regulation of human hydroxysteroid sulfotransferase SULT2A1 by LXR?. Drug Metab Dispos 42:1684-9
Jiang, Mengxi; He, Jinhan; Kucera, Heidi et al. (2014) Hepatic overexpression of steroid sulfatase ameliorates mouse models of obesity and type 2 diabetes through sex-specific mechanisms. J Biol Chem 289:8086-97
He, Jinhan; Hu, Bingfang; Shi, Xiongjie et al. (2013) Activation of the aryl hydrocarbon receptor sensitizes mice to nonalcoholic steatohepatitis by deactivating mitochondrial sirtuin deacetylase Sirt3. Mol Cell Biol 33:2047-55
Jiang, Mengxi; Xie, Wen (2013) Role of the constitutive androstane receptor in obesity and type 2 diabetes: a case study of the endobiotic function of a xenobiotic receptor. Drug Metab Rev 45:156-63
Kittayaruksakul, Suticha; Zhao, Wenchen; Xu, Meishu et al. (2013) Identification of three novel natural product compounds that activate PXR and CAR and inhibit inflammation. Pharm Res 30:2199-208
He, Jinhan; Gao, Jie; Xu, Meishu et al. (2013) PXR ablation alleviates diet-induced and genetic obesity and insulin resistance in mice. Diabetes 62:1876-87
Swanson, Hollie I; Wada, Taira; Xie, Wen et al. (2013) Role of nuclear receptors in lipid dysfunction and obesity-related diseases. Drug Metab Dispos 41:1-11
Chai, Xiaojuan; Zeng, Su; Xie, Wen (2013) Nuclear receptors PXR and CAR: implications for drug metabolism regulation, pharmacogenomics and beyond. Expert Opin Drug Metab Toxicol 9:253-66
Gao, Jie; He, Jinhan; Shi, Xiongjie et al. (2012) Sex-specific effect of estrogen sulfotransferase on mouse models of type 2 diabetes. Diabetes 61:1543-51

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