Potential conflict of interest: Nothing to report.
Autoimmune, Cholestatic and Biliary Disease
G-protein-coupled receptor 30/adenylyl cyclase/protein kinase A pathway is involved in estradiol 17ß-d-glucuronide-induced cholestasis
Article first published online: 21 JAN 2014
© 2014 by the American Association for the Study of Liver Diseases
Volume 59, Issue 3, pages 1016–1029, March 2014
How to Cite
Zucchetti, A. E., Barosso, I. R., Boaglio, A. C., Basiglio, C. L., Miszczuk, G., Larocca, M. C., Ruiz, M. L., Davio, C. A., Roma, M. G., Crocenzi, F. A. and Pozzi, E. J. S. (2014), G-protein-coupled receptor 30/adenylyl cyclase/protein kinase A pathway is involved in estradiol 17ß-d-glucuronide-induced cholestasis. Hepatology, 59: 1016–1029. doi: 10.1002/hep.26752
This work was supported by grants from Agencia Nacional de Promoción Científica y Tecnológica (PICTs 2006 no. 02012 and 2010 no. 1197) and Consejo Nacional de Investigaciones Cientificas y Técnicas (PIP 0691).
- Issue published online: 25 FEB 2014
- Article first published online: 21 JAN 2014
- Accepted manuscript online: 1 OCT 2013 04:10AM EST
- Manuscript Accepted: 16 SEP 2013
- Manuscript Received: 6 MAR 2013
Estradiol-17ß-d-glucuronide (E17G) activates different signaling pathways (e.g., Ca2+-dependent protein kinase C, phosphoinositide 3-kinase/protein kinase B, mitogen-activated protein kinases [MAPKs] p38 and extracellular signal-related kinase 1/2, and estrogen receptor alpha) that lead to acute cholestasis in rat liver with retrieval of the canalicular transporters, bile salt export pump (Abcb11) and multidrug resistance-associated protein 2 (Abcc2). E17G shares with nonconjugated estradiol the capacity to activate these pathways. G-protein-coupled receptor 30 (GPR30) is a receptor implicated in nongenomic effects of estradiol, and the aim of this study was to analyze the potential role of this receptor and its downstream effectors in E17G-induced cholestasis. In vitro, GPR30 inhibition by G15 or its knockdown with small interfering RNA strongly prevented E17G-induced impairment of canalicular transporter function and localization. E17G increased cyclic adenosine monophosphate (cAMP) levels, and this increase was blocked by G15, linking GPR30 to adenylyl cyclase (AC). Moreover, AC inhibition totally prevented E17G insult. E17G also increased protein kinase A (PKA) activity, which was blocked by G15 and AC inhibitors, connecting the links of the pathway, GPR30-AC-PKA. PKA inhibition prevented E17G-induced cholestasis, whereas exchange protein activated directly by cyclic nucleotide/MAPK kinase, another cAMP downstream effector, was not implicated in cAMP cholestatic action. In the perfused rat liver model, inhibition of the GPR30-AC-PKA pathway totally prevented E17G-induced alteration in Abcb11 and Abcc2 function and localization. Conclusion: Activation of GPR30-AC-PKA is a key factor in the alteration of canalicular transporter function and localization induced by E17G. Interaction of E17G with GPR30 may be the first event in the cascade of signaling activation. (Hepatology 2014;59:1016–1029)