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Role of AMP-activated protein kinase α1 in 17α-ethinylestradiol-induced cholestasis in rats

  • Organ Toxicity and Mechanisms
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Abstract

Estrogen-induced cholestasis occurs in many women who are susceptible due to pregnancy or hormone replacement therapy for postmenopausal syndrome. 17α-Ethinylestradiol (EE), as a synthetic estrogen, has been widely used to study the underlying mechanisms of estrogen-induced cholestasis. Recent studies have also reported that liver kinase B1 (LKB1)-mediated activation of AMP-activated protein kinase (AMPK) plays a critical role in the regulation of canalicular network formation. However, the role of AMPK in EE-induced cholestasis remains to be determined. In this study, the effects of EE (1–100 µM) on AMPK activation and the expression of farnesoid X receptor (FXR) and hepatic bile acid transporters were examined in in vitro using 3D-cultured rat primary hepatocytes and in in vivo using rat cholestasis models. We also used specific chemical agonist and antagonist of AMPK, AMPK subunit-specific antibodies and lentiviral shRNAs for AMPKα1 and AMPKα2 to delineate the role of AMPK in EE-induced cholestasis and potential cellular mechanisms. We found that EE-induced phosphorylation of AMPKα1 via extracellular signal-regulated kinases-LKB1-mediated signaling pathways and subsequent nuclear translocation accounted for the down-regulation of FXR and bile acid transporters and disruption of bile acid homeostasis. Inhibition of AMPK activation using an AMPK antagonist Compound C (2 µM) or down-regulation of AMPKα1 using gene-specific shRNA attenuated EE-induced cholestasis both in in vitro and in in vivo. In conclusion, these results revealed that activation of cAMP-ERK-LKB1-AMPKα1 signaling pathway plays a critical role in EE-mediated dysregulation of the expression of FXR and bile acid transporters. AMPKα1 may represent an important therapeutic target for estrogen-induced cholestasis.

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Abbreviations

ICP:

Intrahepatic cholestasis of pregnancy

EE:

17α-Ethinylestradiol

Bsep:

Bile salt export pump

Mrp2:

Multidrug resistance-associated protein 2

Ntcp:

Na+-dependent taurocholate cotransporter

Oatp2:

Organic anion transporters 2

FXR:

Farnesoid X receptor

AMPK:

AMP-activated protein kinase

LKB1:

Liver kinase B1

ERK1/2:

Extracellular signal-regulated kinases

ER:

Estrogen receptor

TCA:

Taurocholate

GPR30:

G protein-coupled receptor 30

CC:

Compound C

GAPDH:

Glyceraldehyde 3-phosphate dehydrogenase

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Acknowledgments

This work was supported by National Natural Science Foundation of China Grants (81320108029 to LZ; 81573514 to ZJ; 81070245, 81270489 to HZ); Specific Fund for Public Interest Research of Traditional Chinese Medicine, Ministry of Finance (201507004-002 to LZ), Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD); National Institutes of Heath Grant R01 DK-057543-11 (to PBH and HZ), VA Merit Awards (to HZ; I01BX001390); VCU Massey Cancer Pilot grant (A35362 to HZ).

Authors contribution

LX, PBH, ZJ, LZ and HZ conceived the original ideas, designed the study, analyzed the data and wrote the manuscript; LX, RL, LL, XC, LS, TW carried out the experiments and data analysis. ZJ and LZ contributed equally.

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Authors and Affiliations

  1. Jiangsu Key Laboratory of Drug Screening, China Pharmaceutical University, Nanjing, 210009, Jiangsu, China

    Xiaojiaoyang Li, Runping Liu, Lan Luo, Linxi Yu, Xin Chen, Lixin Sun, Tao Wang, Zhenzhou Jiang & Luyong Zhang

  2. Department of Microbiology and Immunology, Virginia Commonwealth University, Richmond, VA, 23298, USA

    Runping Liu, Phillip B. Hylemon & Huiping Zhou

  3. Jiangsu Center for Pharmacodynamics Research and Evaluation, China Pharmaceutical University, Nanjing, 210009, Jiangsu, China

    Xiaojiaoyang Li, Tao Wang, Zhenzhou Jiang & Luyong Zhang

  4. McGuire Veterans Affairs Medical Center, Richmond, VA, 23249, USA

    Phillip B. Hylemon & Huiping Zhou

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  1. Xiaojiaoyang Li
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  2. Runping Liu
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Correspondence to Huiping Zhou, Zhenzhou Jiang or Luyong Zhang.

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Li, X., Liu, R., Luo, L. et al. Role of AMP-activated protein kinase α1 in 17α-ethinylestradiol-induced cholestasis in rats. Arch Toxicol 91, 481–494 (2017). https://doi.org/10.1007/s00204-016-1697-8

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  • DOI: https://doi.org/10.1007/s00204-016-1697-8

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