Rho-kinase/myosin Light Chain Kinase Pathway Plays a Key Role in the Impairment of Bile Canaliculi Dynamics Induced by Cholestatic Drugs

Overview

Journal Sci Rep

Specialty Science

Date 2016 May 13

PMID 27169750

Citations 30

Authors

Ahmad Sharanek

Audrey Burban

Matthew Burbank

Remy Le Guevel

Ruoya Li

Andre Guillouzo

Christiane Guguen-Guillouzo

Affiliations

Soon will be listed here.

Abstract

Intrahepatic cholestasis represents a frequent manifestation of drug-induced liver injury; however, the mechanisms underlying such injuries are poorly understood. In this study of human HepaRG and primary hepatocytes, we found that bile canaliculi (BC) underwent spontaneous contractions, which are essential for bile acid (BA) efflux and require alternations in myosin light chain (MLC2) phosphorylation/dephosphorylation. Short exposure to 6 cholestatic compounds revealed that BC constriction and dilation were associated with disruptions in the ROCK/MLCK/myosin pathway. At the studied concentrations, cyclosporine A and chlorpromazine induced early ROCK activity, resulting in permanent MLC2 phosphorylation and BC constriction. However, fasudil reduced ROCK activity and caused rapid, substantial and permanent MLC2 dephosphorylation, leading to BC dilation. The remaining compounds (1-naphthyl isothiocyanate, deoxycholic acid and bosentan) caused BC dilation without modulating ROCK activity, although they were associated with a steady decrease in MLC2 phosphorylation via MLCK. These changes were associated with a common loss of BC contractions and failure of BA clearance. These results provide the first demonstration that cholestatic drugs alter BC dynamics by targeting the ROCK/MLCK pathway; in addition, they highlight new insights into the mechanisms underlying bile flow failure and can be used to identify new predictive biomarkers of drug-induced cholestasis.

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References

Zhou Q, Hennenberg M, Trebicka J, Jochem K, Leifeld L, Biecker E . Intrahepatic upregulation of RhoA and Rho-kinase signalling contributes to increased hepatic vascular resistance in rats with secondary biliary cirrhosis. Gut. 2006; 55(9):1296-305. PMC: 1860046. DOI: 10.1136/gut.2005.081059. View

Herrema H, Czajkowska D, Theard D, van der Wouden J, Kalicharan D, Zolghadr B . Rho kinase, myosin-II, and p42/44 MAPK control extracellular matrix-mediated apical bile canalicular lumen morphogenesis in HepG2 cells. Mol Biol Cell. 2006; 17(7):3291-303. PMC: 1552049. DOI: 10.1091/mbc.e06-01-0067. View

Watanabe T, Hosoya H, Yonemura S . Regulation of myosin II dynamics by phosphorylation and dephosphorylation of its light chain in epithelial cells. Mol Biol Cell. 2006; 18(2):605-16. PMC: 1783795. DOI: 10.1091/mbc.e06-07-0590. View

Cerec V, Glaise D, Garnier D, Morosan S, Turlin B, Drenou B . Transdifferentiation of hepatocyte-like cells from the human hepatoma HepaRG cell line through bipotent progenitor. Hepatology. 2007; 45(4):957-67. DOI: 10.1002/hep.21536. View

Nossaman B, Kadowitz P . The role of the RhoA/rho-kinase pathway in pulmonary hypertension. Curr Drug Discov Technol. 2009; 6(1):59-71. DOI: 10.2174/157016309787581057. View

Stieger B . Role of the bile salt export pump, BSEP, in acquired forms of cholestasis. Drug Metab Rev. 2009; 42(3):437-45. DOI: 10.3109/03602530903492004. View

Fu D, Wakabayashi Y, Ido Y, Lippincott-Schwartz J, Arias I . Regulation of bile canalicular network formation and maintenance by AMP-activated protein kinase and LKB1. J Cell Sci. 2010; 123(Pt 19):3294-302. PMC: 2939801. DOI: 10.1242/jcs.068098. View

Morgan R, Trauner M, van Staden C, Lee P, Ramachandran B, Eschenberg M . Interference with bile salt export pump function is a susceptibility factor for human liver injury in drug development. Toxicol Sci. 2010; 118(2):485-500. DOI: 10.1093/toxsci/kfq269. View

Fu D, Wakabayashi Y, Lippincott-Schwartz J, Arias I . Bile acid stimulates hepatocyte polarization through a cAMP-Epac-MEK-LKB1-AMPK pathway. Proc Natl Acad Sci U S A. 2011; 108(4):1403-8. PMC: 3029747. DOI: 10.1073/pnas.1018376108. View

10.

Padda M, Sanchez M, Akhtar A, Boyer J . Drug-induced cholestasis. Hepatology. 2011; 53(4):1377-87. PMC: 3089004. DOI: 10.1002/hep.24229. View

11.

Antherieu S, Rogue A, Fromenty B, Guillouzo A, Robin M . Induction of vesicular steatosis by amiodarone and tetracycline is associated with up-regulation of lipogenic genes in HepaRG cells. Hepatology. 2011; 53(6):1895-905. DOI: 10.1002/hep.24290. View

12.

Pernelle K, Le Guevel R, Glaise D, Stasio C, Le Charpentier T, Bouaita B . Automated detection of hepatotoxic compounds in human hepatocytes using HepaRG cells and image-based analysis of mitochondrial dysfunction with JC-1 dye. Toxicol Appl Pharmacol. 2011; 254(3):256-66. DOI: 10.1016/j.taap.2011.04.018. View

13.

Dawson S, Stahl S, Paul N, Barber J, Kenna J . In vitro inhibition of the bile salt export pump correlates with risk of cholestatic drug-induced liver injury in humans. Drug Metab Dispos. 2011; 40(1):130-8. DOI: 10.1124/dmd.111.040758. View

14.

Bjornsson E, Jonasson J . Drug-induced cholestasis. Clin Liver Dis. 2013; 17(2):191-209. DOI: 10.1016/j.cld.2012.11.002. View

15.

Antherieu S, Bachour-El Azzi P, Dumont J, Abdel-Razzak Z, Guguen-Guillouzo C, Fromenty B . Oxidative stress plays a major role in chlorpromazine-induced cholestasis in human HepaRG cells. Hepatology. 2012; 57(4):1518-29. DOI: 10.1002/hep.26160. View

16.

Fu D, Mitra K, Sengupta P, Jarnik M, Lippincott-Schwartz J, Arias I . Coordinated elevation of mitochondrial oxidative phosphorylation and autophagy help drive hepatocyte polarization. Proc Natl Acad Sci U S A. 2013; 110(18):7288-93. PMC: 3645550. DOI: 10.1073/pnas.1304285110. View

17.

Wang R, Liu L, Sheps J, Forrest D, Hofmann A, Hagey L . Defective canalicular transport and toxicity of dietary ursodeoxycholic acid in the abcb11-/- mouse: transport and gene expression studies. Am J Physiol Gastrointest Liver Physiol. 2013; 305(4):G286-94. DOI: 10.1152/ajpgi.00082.2013. View

18.

Morgan R, van Staden C, Chen Y, Kalyanaraman N, Kalanzi J, Dunn 2nd R . A multifactorial approach to hepatobiliary transporter assessment enables improved therapeutic compound development. Toxicol Sci. 2013; 136(1):216-41. DOI: 10.1093/toxsci/kft176. View

19.

Pedersen J, Matsson P, Bergstrom C, Hoogstraate J, Noren A, LeCluyse E . Early identification of clinically relevant drug interactions with the human bile salt export pump (BSEP/ABCB11). Toxicol Sci. 2013; 136(2):328-43. PMC: 3858191. DOI: 10.1093/toxsci/kft197. View

20.

Rodrigues A, Lai Y, Cvijic M, Elkin L, Zvyaga T, Soars M . Drug-induced perturbations of the bile acid pool, cholestasis, and hepatotoxicity: mechanistic considerations beyond the direct inhibition of the bile salt export pump. Drug Metab Dispos. 2013; 42(4):566-74. DOI: 10.1124/dmd.113.054205. View