Premium
G‐protein‐coupled receptor 30/adenylyl cyclase/protein kinase A pathway is involved in estradiol 17ß‐ d ‐glucuronide‐induced cholestasis
Author(s) -
Zucchetti Andrés E.,
Barosso Ismael R.,
Boaglio Andrea C.,
Basiglio Cecilia L.,
Miszczuk Gisel,
Larocca M. Cecilia,
Ruiz M. Laura,
Davio Carlos A.,
Roma Marcelo G.,
Crocenzi Fernando A.,
Pozzi Enrique J. Sánchez
Publication year - 2014
Publication title -
hepatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.488
H-Index - 361
eISSN - 1527-3350
pISSN - 0270-9139
DOI - 10.1002/hep.26752
Subject(s) - protein kinase a , adenylyl cyclase , gs alpha subunit , biology , camp dependent pathway , ask1 , gper , microbiology and biotechnology , signal transduction , mitogen activated protein kinase kinase , biochemistry , chemistry , kinase , estrogen receptor , genetics , cancer , breast cancer
Estradiol‐17ß‐ d ‐glucuronide (E17G) activates different signaling pathways (e.g., Ca 2+ ‐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. (H epatology 2014;59:1016–1029)