Abstract
Background
Estradiol-17β-d-glucuronide (E17G) induces cholestasis in vivo, endocytic internalization of the canalicular transporters multidrug resistance-associated protein 2 (Abcc2) and bile salt export pump (Abcb11) being a key pathomechanism. Cyclic AMP (cAMP) prevents cholestasis by targeting these transporters back to the canalicular membrane. In hepatocyte couplets, glucagon and salbutamol, both of which increase cAMP, prevented E17G action by stimulating the trafficking of these transporters by different mechanisms, namely: glucagon activates a protein kinase A-dependent pathway, whereas salbutamol activates an exchange-protein activated by cAMP (Epac)-mediated, microtubule-dependent pathway.
Methods
The present study evaluated whether glucagon and salbutamol prevent E17G-induced cholestasis in a more physiological model, i.e., the perfused rat liver (PRL). Additionally, the preventive effect of in vivo alanine administration, which induces pancreatic glucagon secretion, was evaluated.
Results
In PRLs, glucagon and salbutamol prevented E17G-induced decrease in both bile flow and the secretory activity of Abcc2 and Abcb11. Salbutamol prevention fully depended on microtubule integrity. On the other hand, glucagon prevention was microtubule-independent only at early time periods after E17G administration, but it was ultimately affected by the microtubule disrupter colchicine. Cholestasis was associated with endocytic internalization of Abcb11 and Abcc2, the intracellular carriers being partially colocalized with the endosomal marker Rab11a. This effect was completely prevented by salbutamol, whereas some transporter-containing vesicles remained colocalized with Rab11a after glucagon treatment. In vivo, alanine administration increased hepatic cAMP and accelerated the recovery of bile flow and Abcb11/Abcc2 transport function after E17G administration. The initial recovery afforded by alanine was microtubule-independent, but microtubule integrity was required to sustain this protective effect.
Conclusion
We conclude that modulation of cAMP levels either by direct administration of cAMP modulators or by physiological manipulations leadings to hormone-mediated increase of cAMP levels (alanine administration), prevents estrogen-induced cholestasis in models with preserved liver architecture, through mechanisms similar to those arisen from in vitro studies.
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Abbreviations
- Abcb11:
-
Bile salt export pump
- Abcc2:
-
Multidrug-resistance-associated protein 2
- Ala:
-
Alanine
- E17G:
-
Estradiol 17β-d-glucuronide
- DNP-G:
-
S-(2,4-Dinitrophenyl)-glutathione
- TC:
-
Sodium taurocholate
- Sal:
-
Salbutamol
- Glu:
-
Glucagon
- DMSO:
-
Dimethyl sulfoxide
- PRL:
-
Perfused rat liver
References
Borst P, Elferink RO. Mammalian ABC transporters in health and disease. Annu Rev Biochem. 2002;71:537–592.
Gatmaitan ZC, Arias IM. ATP-dependent transport systems in the canalicular membrane of the hepatocyte. Physiol Rev. 1995;75:261–275.
Esteller A. Physiology of bile secretion. World J Gastroenterol. 2008;14:5641–5649.
Crocenzi FA, Mottino AD, Cao J, et al. Estradiol-17beta-d-glucuronide induces endocytic internalization of Bsep in rats. Am J Physiol Gastrointest Liver Physiol. 2003;285:G449–G459.
Trauner M, Meier PJ, Boyer JL. Molecular regulation of hepatocellular transport systems in cholestasis. J Hepatol. 1999;31:165–178.
Vore M, Liu Y, Huang L. Cholestatic properties and hepatic transport of steroid glucuronides. Drug Metab Rev. 1997;29:183–203.
Adlercreutz H, Tikkanen MJ, Wichmann K, et al. Recurrent jaundice in pregnancy. IV. Quantitative determination of urinary and biliary estrogens, including studies in pruritus gravidarum. J Clin Endocrinol Metab. 1974;38:51–57.
Stieger B, Fattinger K, Madon J, et al. Drug- and estrogen-induced cholestasis through inhibition of the hepatocellular bile salt export pump (Bsep) of rat liver. Gastroenterology. 2000;118:422–430.
Vallejo M, Briz O, Serrano MA, et al. Potential role of trans-inhibition of the bile salt export pump by progesterone metabolites in the etiopathogenesis of intrahepatic cholestasis of pregnancy. J Hepatol. 2006;44:1150–1157.
Mottino AD, Cao J, Veggi LM, et al. Altered localization and activity of canalicular Mrp2 in estradiol-17beta-d-glucuronide-induced cholestasis. Hepatology. 2002;35:1409–1419.
Mottino AD, Crocenzi FA, Pozzi EJ, et al. Role of microtubules in estradiol-17beta-d-glucuronide-induced alteration of canalicular Mrp2 localization and activity. Am J Physiol Gastrointest Liver Physiol. 2005;288:G327–G336.
Roma MG, Crocenzi FA. Sanchez Pozzi EA. Hepatocellular transport in acquired cholestasis: new insights into functional, regulatory and therapeutic aspects. Clin Sci (Lond). 2008;114:567–588.
Kipp H, Arias IM. Trafficking of canalicular ABC transporters in hepatocytes. Annu Rev Physiol. 2002;64:595–608.
Roelofsen H, Soroka CJ, Keppler D, et al. Cyclic AMP stimulates sorting of the canalicular organic anion transporter (Mrp2/cMoat) to the apical domain in hepatocyte couplets. J Cell Sci. 1998;111:1137–1145.
Steinberg SF, Chow YK, Bilezikian JP. Cyclic AMP and guanine nucleotide regulatory proteins. In: Arias IM, Jakoby WB, Popper H, eds. The Liver: Biology and Pathobiology. 2nd ed. New York: Raven Press, Ltd; 2003:769–776.
Branum GD, Bowers BA, Watters CR, et al. Biliary response to glucagon in humans. Ann Surg. 1991;213:335–340.
Morgan NG, Blackmore PF, Exton JH. Age-related changes in the control of hepatic cyclic AMP levels by alpha 1- and beta 2-adrenergic receptors in male rats. J Biol Chem. 1983;258:5103–5109.
Bollen M, Keppens S, Stalmans W. Specific features of glycogen metabolism in the liver. Biochem J. 1998;336:19–31.
Zucchetti AE, Barosso IR, Boaglio A, et al. Prevention of estradiol 17beta-d-glucuronide-induced canalicular transporter internalization by hormonal modulation of cAMP in rat hepatocytes. Mol Biol Cell. 2011;22:3902–3915.
Crocenzi FA. Sanchez Pozzi EJ, Ruiz ML et al. Ca(2 +)-dependent protein kinase C isoforms are critical to estradiol 17beta-d-glucuronide-induced cholestasis in the rat. Hepatology. 2008;48:1885–1895.
Hinchman CA, Matsumoto H, Simmons TW, et al. Intrahepatic conversion of a glutathione conjugate to its mercapturic acid. Metabolism of 1-chloro-2,4-dinitrobenzene in isolated perfused rat and guinea pig livers. J Biol Chem. 1991;266:22179–22185.
Talalay P. Enzymic analysis of steroid hormones. Methods Biochem Anal. 1960;8:119–143.
Muller WA, Faloona GR, Unger RH. The effect of alanine on glucagon secretion. J Clin Invest. 1971;50:2215–2218.
Porcellati F, Pampanelli S, Rossetti P, et al. Effect of the amino acid alanine on glucagon secretion in non-diabetic and type 1 diabetic subjects during hyperinsulinaemic euglycaemia, hypoglycaemia and post-hypoglycaemic hyperglycaemia. Diabetologia. 2007;50:422–430.
Rocha DM, Faloona GR, Unger RH. Glucagon-stimulating activity of 20 amino acids in dogs. J Clin Invest. 1972;51:2346–2351.
Tanaka K, Inoue S, Saito S, et al. Hepatic vagal amino acid sensors modulate amino acid-induced insulin and glucagon secretion in the rat. J Auton Nerv Syst. 1993;42:225–231.
Jendrassik J, Grof P. Vereinfachte photometrische Methode zur Bestimmung des Blut-bilirubins. Biochem Z. 1938;297:81–89.
Davio CA, Cricco GP, Bergoc RM, et al. H1 and H2 histamine receptors in N-nitroso-N-methylurea (NMU)-induced carcinomas with atypical coupling to signal transducers. Biochem Pharmacol. 1995;50:91–96.
Sabbatini ME, Villagra A, Davio CA, et al. Atrial natriuretic factor stimulates exocrine pancreatic secretion in the rat through NPR-C receptors. Am J Physiol Gastrointest Liver Physiol. 2003;285:G929–G937.
Lowry OH, Rosembrough NJ, Farr AL, et al. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951;193:265–275.
Wakabayashi Y, Arias IM. Apical recycling of canalicular ABC transporters. In: Arias IM, Wolkoff A, Boyer JM, Shafritz D, Fausto N, Harvey A, eds. The Liver: Biology and Pathobiology, 5. Chichester: Wiley; 2009:349–358.
Wakabayashi Y, Lippincott-Schwartz J, Arias IM. Intracellular trafficking of bile salt export pump (ABCB11) in polarized hepatic cells: constitutive cycling between the canalicular membrane and rab11-positive endosomes. Mol Biol Cell. 2004;15:3485–3496.
Wang W, Soroka CJ, Mennone A, et al. Radixin is required to maintain apical canalicular membrane structure and function in rat hepatocytes. Gastroenterology. 2006;131:878–884.
Huang L, Smit JW, Meijer DK, et al. Mrp2 is essential for estradiol-17beta(beta-d-glucuronide)-induced cholestasis in rats. Hepatology. 2000;32:66–72.
Acknowledgments
We deeply thank Dr. Carlos Davio (Cátedra de Química Medicinal, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires) for his expert advice and guidance in cAMP measurements. This work was supported by grants from Agencia Nacional de Promoción Científica y Tecnológica (PICTs 2006 No 02012 and 2010 No 1197), Consejo Nacional de Investigaciones Científicas y Técnicas (PIP 0691), and Fundación Allende.
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Zucchetti, A.E., Barosso, I.R., Boaglio, A.C. et al. Hormonal Modulation of Hepatic cAMP Prevents Estradiol 17β-d-Glucuronide-Induced Cholestasis in Perfused Rat Liver. Dig Dis Sci 58, 1602–1614 (2013). https://doi.org/10.1007/s10620-013-2558-4
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DOI: https://doi.org/10.1007/s10620-013-2558-4