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Archives of Toxicology

, Volume 92, Issue 2, pp 729–744 | Cite as

Activation of insulin-like growth factor 1 receptor participates downstream of GPR30 in estradiol-17β-d-glucuronide-induced cholestasis in rats

  • Ismael R. Barosso
  • Gisel S. Miszczuk
  • Nadia Ciriaci
  • Romina B. Andermatten
  • Paula M. Maidagan
  • Valeria Razori
  • Diego R. Taborda
  • Marcelo G. Roma
  • Fernando A. Crocenzi
  • Enrique J. Sánchez PozziEmail author
Molecular Toxicology

Abstract

Estradiol-17β-d-glucuronide (E17G), through the activation of different signaling proteins, induces acute endocytic internalization of canalicular transporters in rat, including multidrug resistance-associated protein 2 (Abcc2) and bile salt export pump (Abcb11), generating cholestasis. Insulin-like growth factor 1 receptor (IGF-1R) is a membrane-bound tyrosine kinase receptor that can potentially interact with proteins activated by E17G. The aim of this study was to analyze the potential role of IGF-1R in the effects of E17G in isolated perfused rat liver (IPRL) and isolated rat hepatocyte couplets. In vitro, IGF-1R inhibition by tyrphostin AG1024 (TYR, 100 nM), or its knock-down with siRNA, strongly prevented E17G-induced impairment of Abcc2 and Abcb11 function and localization. The protection by TYR was not additive to that produced by wortmannin (PI3K inhibitor, 100 nM), and both protections share the same dependency on microtubule integrity, suggesting that IGF-1R shared the signaling pathway of PI3K/Akt. Further analysis of the activation of Akt and IGF-1R induced by E17G indicated a sequence of activation GPR30-IGF-1R-PI3K/Akt. In IPRL, an intraportal injection of E17G triggered endocytosis of Abcc2 and Abcb11, and this was accompanied by a sustained decrease in the bile flow and the biliary excretion of Abcc2 and Abcb11 substrates. TYR did not prevent the initial decay, but it greatly accelerated the recovery to normality of these parameters and the reinsertion of transporters into the canalicular membrane. In conclusion, the activation of IGF-1R is a key factor in the alteration of canalicular transporter function and localization induced by E17G, and its activation follows that of GPR30 and precedes that of PI3K/Akt.

Keywords

Abcb11 Abcc2 IGF-1 receptor Cholestasis ABC transporters 

Abbreviations

Abcc2

Multidrug resistance-associated protein 2

Abcb11

Bile salt export pump

E17G

Estradiol 17β-d-glucuronide

EGFR

Epidermal growth factor receptor

ERα

Estrogen receptor alpha

GPR30

G protein-coupled receptor 30

IGF

Insulin-like growth factor 1

PI3K

Phosphoinositide 3-kinase

Akt

Protein kinase B

CMFDA

5-Chloromethylfluorescein diacetate

GS-MF

Glutathione methylfluorescein

CGamf

Cholyl-glycylamido-fluorescein

DMSO

Dimethyl sulfoxide

IRHC

Isolated rat hepatocyte couplets

SCRH

Sandwich-cultured rat hepatocytes

cVA

Canalicular vacuolar accumulation

IPRL

Isolated perfused rat liver

Notes

Acknowledgements

We thank J. Pellegrino for assistance with confocal microscopy.

Compliance with ethical standards

Financial support

This work was supported by grants from Agencia Nacional de Promoción Científica y Tecnológica (PICTs 2013 N° 1222 and 2013 N° 0974) and Consejo Nacional de Investigaciones Científicas y Técnicas (PIP 0217 and PUE IFISE 0089).

Supplementary material

204_2017_2098_MOESM1_ESM.eps (170 kb)
Additional Fig. 1. Activation studies of insulin-like growth factor receptor-I (IGF-IR). IRHCs were incubated with IGF-I alone (0.1-20 nM) (a) and together with the EGFR agonist EGF (b); cVA of GS-MF was calculated as the percentage of couplets displaying visible fluorescence in their canalicular vacuoles from a total of at least 200 couplets per preparation, referred to as control cVA values. Data are expressed as mean ± SEM (n = 3). * Significantly different of control (EPS 169 kb)
204_2017_2098_MOESM2_ESM.eps (2 mb)
Additional Fig. 2. Image analysis of Abcc2 and Abcb11 in IRHC. a. Distributions of Abcc2 (a, left) and actin (a, right) fluorescence intensity. b. Distributions of Abcb11 (b, left) and actin (b, right) fluorescence intensity. The densitometric analysis of the distribution of Abcc2 and Abcb11 fluorescence intensity was performed along an 8-μm line perpendicular to the canalicular vacuole (4 µM to each side of the vacuole center) using the ImageJ 1.48 software (National Institutes of Health, Bethesda, MD) with the RGB profile plot plugin. F-actin-associated fluorescence was used to delimit the canalicular space. Each line profile measurement was normalized to the sum of all intensities of the respective measurement. The distribution of transporter-associated fluorescence (green channel), expressed as a percentage of the total, was then calculated for each canaliculi and compared statistically using the Mann–Whitney test; any difference among groups thus reflects changes in localization along the 8-μm line. Analysis of confocal microscopy data was performed in a blinded manner. Results are expressed as mean ± SEM. n = 6–8 canalicular vacuoles per preparation, from three independent preparations. Statistical analysis of the profiles revealed a significant internalization of Abcc2 and Abcb11 under E17G treatment (p < 0.05 vs control), which was completely abolished by TYR (p < 0.05 vs E17G). Note that none of the treatments affected the normal distribution of actin, which showed similar profile (EPS 2079 kb)
204_2017_2098_MOESM3_ESM.eps (2.3 mb)
Additional Fig. 3. Image analysis of Abcc2 and Abcb11 in PRLs. Distributions of Abcc2 (a, left)- and actin (a, right)-associated fluorescence intensity. b. Distributions of Abcb11 (b, left)- and actin (b, right)-associated fluorescence intensity. Graphs represent the intensity of fluorescence associated with the transporters along an 8-µm line (from -4 µm to +4 µm of the canalicular center) perpendicular to the canaliculus. Occludin-associated fluorescence was used to delimit the canalicular space. The variances of the densitometric profiles of Abcb11 and Abcc2 localization were compared with the Mann–Whitney U test. In control livers, transporter-associated fluorescence was concentrated in the canalicular space. E17G-induced internalization of transporters from the canalicular membrane (P < 0.01 versus control) was detected as a decrease in the fluorescence intensity in the canalicular area together with an increased fluorescence at a greater distance from the canaliculus. Distribution profiles of livers treated with E17G+TYR were similar to control and indicated a significantly decreased of Abcb11 and Abcc2 internalization (P < 0.01 versus E17G). (n = 20-50 canaliculi per preparation, three independent preparations). Statistical analysis of the distribution profiles of occludin showed no changes in the normal distribution by any of the treatments (EPS 2384 kb)

References

  1. Adlercreutz H, Tikkanen MJ, Wichmann K et al (1974) Recurrent jaundice in pregnancy. IV. Quantitative determination of urinary and biliary estrogens, including studies in pruritus gravidarum. J Clin Endocrinol Metab 38:51–57. doi: 10.1210/jcem-38-1-51 CrossRefPubMedGoogle Scholar
  2. Barosso IR, Zucchetti AE, Boaglio AC et al (2012) Sequential activation of classic PKC and estrogen receptor α is involved in estradiol 17β-d-glucuronide-induced cholestasis. PLoS One 7:e50711. doi: 10.1371/journal.pone.0050711 CrossRefPubMedPubMedCentralGoogle Scholar
  3. Barosso IR, Zucchetti AE, Miszczuk GS et al (2015) EGFR participates downstream of ERα in estradiol-17β-d-glucuronide-induced impairment of Abcc2 function in isolated rat hepatocyte couplets. Arch Toxicol 90:891–903. doi: 10.1007/s00204-015-1507-8 CrossRefPubMedGoogle Scholar
  4. Boaglio AC, Zucchetti AE, Sanchez Pozzi EJ et al (2010) Phosphoinositide 3-kinase/protein kinase B signaling pathway is involved in estradiol 17beta-d-glucuronide-induced cholestasis: complementarity with classical protein kinase C. Hepatology 52:1465–1476CrossRefPubMedGoogle Scholar
  5. Boaglio AC, Zucchetti AE, Toledo FD et al (2012) ERK1/2 and p38 MAPKs are complementarily involved in estradiol 17{β}-d-glucuronide-induced cholestasis: crosstalk with cPKC and PI3K. PLoS One 7:e49255. doi: 10.1371/journal.pone.0049255 CrossRefPubMedPubMedCentralGoogle Scholar
  6. Borst P, Elferink RO (2002) Mammalian ABC transporters in health and disease. Annu Rev Biochem 71:537–592. doi: 10.1146/annurev.biochem.71.102301.093055 CrossRefPubMedGoogle Scholar
  7. Crocenzi FA, Mottino AD, Sánchez Pozzi EJ et al (2003a) Impaired localisation and transport function of canalicular Bsep in taurolithocholate induced cholestasis in the rat. Gut 52:1170–1177CrossRefPubMedPubMedCentralGoogle Scholar
  8. Crocenzi FA, Sanchez Pozzi EJ, Pellegrino JM et al (2003b) Preventive effect of silymarin against taurolithocholate-induced cholestasis in the rat. Biochem Pharmacol 66:355–364CrossRefPubMedGoogle Scholar
  9. Crocenzi FA, Sanchez Pozzi EJ, Ruiz ML et al (2008) Ca(2+)-dependent protein kinase C isoforms are critical to estradiol 17beta-d-glucuronide-induced cholestasis in the rat. Hepatology 48:1885–1895CrossRefPubMedPubMedCentralGoogle Scholar
  10. Donner MG, Schumacher S, Warskulat U et al (2007) Obstructive cholestasis induces TNF-alpha- and IL-1 -mediated periportal downregulation of Bsep and zonal regulation of Ntcp, Oatp1a4, and Oatp1b2. Am J Physiol Gastrointest Liver Physiol 293(6):G1134–G1146CrossRefPubMedGoogle Scholar
  11. Esteller A (2008) Physiology of bile secretion. World J Gastroenterol 14:5641–5649CrossRefPubMedPubMedCentralGoogle Scholar
  12. Franco C, Bengtsson B-A, Johannsson G (2006) The GH/IGF-1 axis in obesity: physiological and pathological aspects. Metab Syndr Relat Disord 4:51–56. doi: 10.1089/met.2006.4.51 CrossRefPubMedGoogle Scholar
  13. Garcia F, Kierbel A, Larocca MC et al (2001) The water channel aquaporin-8 is mainly intracellular in rat hepatocytes, and its plasma membrane insertion is stimulated by cyclic AMP. J Biol Chem 276:12147–12152CrossRefPubMedGoogle Scholar
  14. García-Regalado A, Guzmán-Hernández ML, Ramírez-Rangel I et al (2008) G protein-coupled receptor-promoted trafficking of Gbeta1gamma2 leads to AKT activation at endosomes via a mechanism mediated by Gbeta1gamma2-Rab11a interaction. Mol Biol Cell 19:4188–4200. doi: 10.1091/mbc.E07-10-1089 CrossRefPubMedPubMedCentralGoogle Scholar
  15. Garcia-Segura LM, Arévalo M-A, Azcoitia I (2010) Interactions of estradiol and insulin-like growth factor-I signalling in the nervous system. In: Progress in brain research. pp 251–272Google Scholar
  16. Gatmaitan ZC, Arias IM (1995) ATP-dependent transport systems in the canalicular membrane of the hepatocyte. Physiol Rev 75:261–275CrossRefPubMedGoogle Scholar
  17. Gautam A, Ng OC, Boyer JL (1987) Isolated rat hepatocyte couplets in short-term culture: structural characteristics and plasma membrane reorganization. Hepatology 7:216–223CrossRefPubMedGoogle Scholar
  18. Kato H, Faria TN, Stannard B et al (1994) Essential role of tyrosine residues 1131, 1135, and 1136 of the insulin-like growth factor-I (IGF-I) receptor in IGF-I action. Mol Endocrinol 8:40–50. doi: 10.1210/mend.8.1.7512194 PubMedGoogle Scholar
  19. Kuemmerle JF, Murthy KS (2001) Coupling of the insulin-like growth factor-I receptor tyrosine kinase to Gi2 in human intestinal smooth muscle: G-dependent mitogen-activated protein kinase activation and growth. J Biol Chem 276:7187–7194. doi: 10.1074/jbc.M011145200 CrossRefPubMedGoogle Scholar
  20. Lappano R, De Marco P, De Francesco EM et al (2013) Cross-talk between GPER and growth factor signaling. J Steroid Biochem Mol Biol 137:50–56. doi: 10.1016/j.jsbmb.2013.03.005 CrossRefPubMedGoogle Scholar
  21. Maglova LM, Jackson AM, Meng XJ et al (1995) Transport characteristics of three fluorescent conjugated bile acid analogs in isolated rat hepatocytes and couplets. Hepatology 22:637–647PubMedGoogle Scholar
  22. Misra S, Varticovski L, Arias IM (2003) Mechanisms by which cAMP increases bile acid secretion in rat liver and canalicular membrane vesicles. Am J Physiol Gastrointest Liver Physiol 285(2):G316–G324CrossRefPubMedGoogle Scholar
  23. Miszczuk GS, Barosso IR, Zucchetti AE et al (2015) Sandwich-cultured rat hepatocytes as an in vitro model to study canalicular transport alterations in cholestasis. Arch Toxicol 89:979–990. doi: 10.1007/s00204-014-1283-x CrossRefPubMedGoogle Scholar
  24. Miyata M (2004) Role of farnesoid X receptor in the enhancement of canalicular bile acid output and excretion of unconjugated bile acids: a mechanism for protection against cholic acid-induced liver toxicity. J Pharmacol Exp Ther 312:759–766. doi: 10.1124/jpet.104.076158 CrossRefPubMedGoogle Scholar
  25. Mottino AD, Hoffman T, Jennes L et al (2001) Expression of multidrug resistance-associated protein 2 in small intestine from pregnant and postpartum rats. Am J Physiol Gastrointest Liver Physiol 280:G1261–G1273CrossRefPubMedGoogle Scholar
  26. Mottino AD, Cao J, Veggi LM et al (2002) Altered localization and activity of canalicular Mrp2 in estradiol-17β-d-glucuronide-induced cholestasis. Hepatology 35:1409–1419CrossRefPubMedGoogle Scholar
  27. Mottino AD, Veggi LM, Wood M et al (2003) Biliary secretion of glutathione in estradiol 17beta-d-glucuronide-induced cholestasis. J Pharmacol Exp Ther 307:306–313CrossRefPubMedGoogle Scholar
  28. Mottino AD, Crocenzi FA, Pozzi EJS et al (2005) Role of microtubules in estradiol-17beta-d-glucuronide-induced alteration of canalicular Mrp2 localization and activity. Am J Physiol Gastrointest Liver Physiol 288:G327–G336CrossRefPubMedGoogle Scholar
  29. Párrizas M, Gazit A, Levitzki A et al (1997) Specific inhibition of insulin-like growth factor-1 and insulin receptor tyrosine kinase activity and biological function by tyrphostins. Endocrinology 138:1427–1433. doi: 10.1210/endo.138.4.5092 CrossRefPubMedGoogle Scholar
  30. Pfaffl MW, Horgan GW, Dempfle L (2002) Relative expression software tool (REST) for group-wise comparison and statistical analysis of relative expression results in real-time PCR. Nucleic Acids Res 30:e36CrossRefPubMedPubMedCentralGoogle Scholar
  31. Qadir XV, Chen W, Han C et al (2015) miR-223 Deficiency protects against Fas-induced hepatocyte apoptosis and liver injury through targeting insulin-like growth factor 1 receptor. Am J Pathol 185:3141–3151. doi: 10.1016/j.ajpath.2015.08.020 CrossRefPubMedPubMedCentralGoogle Scholar
  32. Roma MG, Milkiewicz P, Elias E, Coleman R (2000) Control by signaling modulators of the sorting of canalicular transporters in rat hepatocyte couplets: role of the cytoskeleton. Hepatology 32:1342–1356CrossRefPubMedGoogle Scholar
  33. Song RX-D, Zhang Z, Chen Y et al (2007) Estrogen signaling via a linear pathway involving insulin-like growth factor I receptor, matrix metalloproteinases, and epidermal growth factor receptor to activate mitogen-activated protein kinase in MCF-7 breast cancer cells. Endocrinology 148:4091–4101. doi: 10.1210/en.2007-0240 CrossRefPubMedPubMedCentralGoogle Scholar
  34. Suleiman M, Singh R, Stewart C (2007) Apoptosis and the cardiac action of insulin-like growth factor I. Pharmacol Ther 114:278–294. doi: 10.1016/j.pharmthera.2007.03.001 CrossRefPubMedGoogle Scholar
  35. Troncoso R, Ibarra C, Vicencio JM et al (2014) New insights into IGF-1 signaling in the heart. Trends Endocrinol Metab 25:128–137CrossRefPubMedGoogle Scholar
  36. Vore M (1987) Estrogen cholestasis. Membranes, metabolites, or receptors? Gastroenterology 93:643–649CrossRefPubMedGoogle Scholar
  37. Vore M, Liu Y, Huang L (1997) Cholestatic properties and hepatic transport of steroid glucuronides. Drug Metabol Rev 29:183–203CrossRefGoogle Scholar
  38. Wang L, Soroka CJ, Boyer JL (2002) The role of bile salt export pump mutations in progressive familial intrahepatic cholestasis type II. J Clin Invest 110:965–972CrossRefPubMedPubMedCentralGoogle Scholar
  39. Wilton JC, Williams DE, Strain AJ et al (1991) Purification of hepatocyte couplets by centrifugal elutriation. Hepatology 14:180–183CrossRefPubMedGoogle Scholar
  40. Yuan B, Latek R, Hossbach M et al (2004) siRNA Selection Server: an automated siRNA oligonucleotide prediction server. Nucleic Acids Res 32:W130–W134. doi: 10.1093/nar/gkh366 CrossRefPubMedPubMedCentralGoogle Scholar
  41. Zheng H, Worrall C, Shen H et al (2012) Selective recruitment of G protein-coupled receptor kinases (GRKs) controls signaling of the insulin-like growth factor 1 receptor. Proc Natl Acad Sci USA 109:7055–7060. doi: 10.1073/pnas.1118359109 CrossRefPubMedPubMedCentralGoogle Scholar
  42. Zucchetti AE, Barosso IR, Boaglio A et al (2011) Prevention of estradiol 17beta-d-glucuronide-induced canalicular transporter internalization by hormonal modulation of cAMP in rat hepatocytes. Mol Biol Cell 22:3902–3915. doi: 10.1091/mbc.E11-01-0047 CrossRefPubMedPubMedCentralGoogle Scholar
  43. Zucchetti AE, Barosso IR, Boaglio AC et al (2013) Hormonal modulation of hepatic cAMP prevents estradiol 17beta-d-glucuronide-induced cholestasis in perfused rat liver. Dig Dis Sci 58:1602–1614. doi: 10.1007/s10620-013-2558-4 CrossRefPubMedGoogle Scholar
  44. Zucchetti AE, Barosso IR, Boaglio AC et al (2014) G-protein-coupled receptor 30/adenylyl cyclase/protein kinase A pathway is involved in estradiol 17{β}-d-glucuronide-induced cholestasis. Hepatology 59:1016–1029. doi: 10.1002/hep.26752 CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany 2017

Authors and Affiliations

  • Ismael R. Barosso
    • 1
  • Gisel S. Miszczuk
    • 1
  • Nadia Ciriaci
    • 1
  • Romina B. Andermatten
    • 1
  • Paula M. Maidagan
    • 1
  • Valeria Razori
    • 1
  • Diego R. Taborda
    • 1
  • Marcelo G. Roma
    • 1
  • Fernando A. Crocenzi
    • 1
  • Enrique J. Sánchez Pozzi
    • 1
    Email author
  1. 1.Instituto de Fisiología Experimental (IFISE), Facultad de Ciencias Bioquímicas y Farmacéuticas (CONICET-U.N.R.)RosarioArgentina

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