Hormesis in Cholestatic Liver Disease; Preconditioning with Low Bile Acid Concentrations Protects against Bile Acid-Induced Toxicity

Esther M. Verhaag, Manon Buist-Homan, Martijn Koehorst, Albert K. Groen, Han Moshage, Klaas Nico Faber*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

8 Citations (Scopus)
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Abstract

Introduction

Cholestasis is characterized by accumulation of bile acids and inflammation, causing hepatocellular damage. Still, liver damage markers are highest in acute cholestasis and drop when this condition becomes chronic, indicating that hepatocytes adapt towards the hostile environment. This may be explained by a hormetic response in hepatocytes that limits cell death during cholestasis.

Aim

To investigate the mechanisms that underlie the hormetic response that protect hepatocytes against experimental cholestatic conditions.

Methods

HepG2.rNtcp cells were preconditioned (24 h) with sub-apoptotic concentrations (0.1-50 mu M) of various bile acids, the superoxide donor menadione, TNF-alpha or the Farsenoid X Receptor agonist GW4064, followed by a challenge with the apoptosis-inducing bile acid glycochenodeoxycholic acid (GCDCA; 200 mu M for 4 h), menadione (50 mu M, 6 h) or cytokine mixture (CM; 6 h). Levels of apoptotic and necrotic cell death, mRNA expression of the bile salt export pump (ABCB11) and bile acid sensors, as well as intracellular GCDCA levels were analyzed.

Results

Preconditioning with the pro-apoptotic bile acids GCDCA, taurocholic acid, or the protective bile acids (tauro) ursodeoxycholic acid reduced GCDCA-induced caspase-3/7 activity in HepG2.rNtcp cells. Bile acid preconditioning did not induce significant levels of necrosis in GCDCA-challenged HepG2.rNtcp cells. In contrast, preconditioning with cholic acid, menadione or TNF-alpha potentiated GCDCA-induced apoptosis. GCDCA preconditioning specifically reduced GCDCA-induced cell death and not CM-or menadione-induced apoptosis. The hormetic effect of GCDCA preconditioning was concentration-and time-dependent. GCDCA-, CDCA-and GW4064-preconditioning enhanced ABCB11 mRNA levels, but in contrast to the bile acids, GW4064 did not significantly reduce GCDCA-induced caspase-3/7 activity. The GCDCA challenge strongly increased intracellular levels of this bile acid, which was not lowered by GCDCA-preconditioning.

Conclusions

Sub-toxic concentrations of bile acids in the range that occur under normal physiological conditions protect HepG2.rNtcp cells against GCDCA-induced apoptosis, which is independent of FXR-controlled changes in bile acid transport.

Original languageEnglish
Article numbere0149782
Number of pages16
JournalPLoS ONE
Volume11
Issue number3
DOIs
Publication statusPublished - 7-Mar-2016

Keywords

  • ISOLATED RAT HEPATOCYTES
  • FARNESOID-X-RECEPTOR
  • SALT EXPORT PUMP
  • REPERFUSION INJURY
  • INDUCED APOPTOSIS
  • OBSTRUCTIVE CHOLESTASIS
  • MOLECULAR-MECHANISMS
  • OXIDATIVE STRESS
  • ACTIVATION
  • FXR

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