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Adenoviral transfer of human aquaporin -1 gene to rat liver improves bile flow in estrogen-induced cholestasis


Estrogens can cause liver cholestatic disease. As downregulation of hepatocyte canalicular aquaporin-8 (AQP8) water channels has been involved in estrogen-induced bile secretory failure, we tested whether the archetypal water channel AQP1 improves 17α-ethinylestradiol (EE)-induced cholestasis. EE administration to rats reduced bile flow by 50%. A recombinant adenoviral (Ad) vector encoding human AQP1 (hAQP1), AdhAQP1, or a control vector was administered by retrograde bile ductal infusion. Hepatocyte canalicular hAQP1 expression was confirmed by liver immunostaining and immunoblotting in purified membrane fractions. Accordingly, canalicular osmotic water permeability was markedly increased. Bile flow, either basal or bile salt-stimulated was significantly augmented by over 50%. The choleretic efficiency of endogenous bile salts (that is, volume of bile per μmol of excreted bile salt) was significantly increased by 45% without changes in the biliary bile salt composition. Our data suggest that the adenoviral transfer of hAQP1 gene to the livers of EE-induced cholestatic rats improves bile flow by enhancing the AQP-mediated bile salt-induced canalicular water secretion. This novel finding might have potential therapeutic implications for cholestatic diseases.

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  1. Lee J, Boyer JL . Molecular alterations in hepatocyte transport mechanisms in acquired cholestatic liver disorders. Semin Liver Dis 2000; 20: 373–384.

    Article  CAS  Google Scholar 

  2. Crocenzi FA, Sánchez Pozzi EJ, Pellegrino JM, Favre CO, Rodríguez Garay EA, Mottino AD et al. Beneficial effects of silymarin on estrogen-induced cholestasis in the rat: a study in vivo and in isolated hepatocyte couplets. Hepatology 2001; 34: 329–339.

    Article  CAS  Google Scholar 

  3. King LS, Kozono D, Agre P . From structure to disease: the evolving tale of aquaporin biology. Nat Rev Mol Cell Biol 2004; 5: 687–698.

    Article  CAS  Google Scholar 

  4. Marinelli RA, Lehmann GL, Soria LR, Marchissio MJ . Hepatocyte aquaporins in bile formation and cholestasis. Front Biosci (Landmark Ed) 2011; 16: 2642–2652.

    Article  CAS  Google Scholar 

  5. Portincasa P, Calamita G . Water channel proteins in bile formation and flow in health and disease: when immiscible becomes miscible. Mol Aspects Med 2012; 33: 651–664.

    Article  CAS  Google Scholar 

  6. Carreras FI, Lehmann GL, Ferri D, Tioni MF, Calamita G, Marinelli RA . Defective hepatocyte aquaporin-8 expression and reduced canalicular membrane water permeability in estrogen-induced cholestasis. Am J Physiol Gastrointest Liver Physiol 2007; 292: G905–G912.

    Article  CAS  Google Scholar 

  7. Delporte C, O'Connell BC, He X, Lancaster HE, O'Connell AC, Agre P et al. Increased fluid secretion after adenoviral-mediated transfer of the aquaporin-1 cDNA to irradiated rat salivary glands. Proc Natl Acad Sci USA 1997; 94: 3268–3273.

    Article  CAS  Google Scholar 

  8. Shan Z, Li J, Zheng C, Liu X, Fan Z, Zhang C et al. Increased fluid secretion after adenoviral-mediated transfer of the human aquaporin-1 cDNA to irradiated miniature pig parotid glands. Mol Ther 2005; 11: 444–451.

    Article  CAS  Google Scholar 

  9. Baum BJ, Alevizos I, Zheng C, Cotrim AP, Liu S, McCullagh L et al. Early responses to adenoviral-mediated transfer of the aquaporin-1 cDNA for radiation-induced salivary hypofunction. Proc Natl Acad Sci USA 2012; 109: 19403–19407.

    Article  CAS  Google Scholar 

  10. Marinelli RA, Tietz PS, Pham LD, Rueckert L, Agre P, LaRusso NF . Secretin induces the apical insertion of aquaporin-1 water channels in rat cholangiocytes. Am J Physiol 1999; 276: G280–G286.

    CAS  Google Scholar 

  11. Terao R, Honda K, Hatano E, Uehara T, Yamamoto M, Yamaoka Y . Suppression of proliferative cholangitis in a rat model with direct adenovirus-mediated retinoblastoma gene transfer to the biliary tract. Hepatology 1998; 28: 605–612.

    Article  CAS  Google Scholar 

  12. Kuriyama S, Yoshiji H, Nakai S, Deguchi A, Uchida N, Kimura Y et al. Adenovirus-mediated gene transfer into rat livers: comparative study of retrograde intrabiliary and antegrade intraportal administration. Oncol Rep 2005; 13: 69–74.

    CAS  PubMed  Google Scholar 

  13. Monte MJ, Badia MD, Palomero F, el-Mir MY, Alonso JR, Marin JJ . Effects of selective zonal injury on bile acid-induced bile flow in the isolated rat liver. Am J Physiol 1993; 264: G1103–G1111.

    CAS  PubMed  Google Scholar 

  14. Zheng C, Goldsmith CM, Mineshiba F, Chiorini JA, Kerr A, Wenk ML et al. Toxicity and biodistribution of a first-generation recombinant adenoviral vector, encoding aquaporin-1, after retroductal delivery to a single rat submandibular gland. Hum Gene Ther 2006; 17: 1122–1133.

    Article  CAS  Google Scholar 

  15. Jiang X, Ren Y, Williford JM, Li Z, Mao HQ . Liver-targeted gene delivery through retrograde intrabiliary infusion. In: Manfred O, David O (eds) Nanotechnology for Nucleic Acid Delivery: Methods and Protocols, Methods Mol Biol. Humana Press: New Jersey, 2013, pp 275–284.

    Chapter  Google Scholar 

  16. Tominaga K, Kuriyama S, Yoshiji H, Deguchi A, Kita Y, Funakoshi F et al. Repeated adenoviral administration into the biliary tract can induce repeated expression of the original gene construct in rat livers without immunosuppressive strategies. Gut 2004; 53: 1167–117.

    Article  CAS  Google Scholar 

  17. Jessner W, Zsembery A, Graf J . Transcellular water transport in hepatobiliary secretion and role of aquaporins in liver. Wien Med Wochenschr 2008; 158: 565–569.

    Article  Google Scholar 

  18. Arrese M, Macias RI, Briz O, Perez MJ, Marin JJ . Molecular pathogenesis of intrahepatic cholestasis of pregnancy. Expert Rev Mol Med 2008; 10: e9.

    Article  Google Scholar 

  19. Pauli-Magnus C, Meier PJ, Stieger B . Genetic determinants of drug-induced cholestasis and intrahepatic cholestasis of pregnancy. Semin Liver Dis 2010; 30: 147–159.

    Article  CAS  Google Scholar 

  20. Hung DY, Siebert GA, Chang P, Roberts MS . Hepatic pharmacokinetics of taurocholate in the normal and cholestatic rat liver. Br J Pharmacol 2005; 145: 57–65.

    Article  CAS  Google Scholar 

  21. Koopen NR, Wolters H, Havinga R, Vonk RJ, Jansen PL, Müller M et al. Impaired activity of the bile canalicular organic anion transporter (Mrp2/cmoat) is not the main cause of ethinylestradiol-induced cholestasis in the rat. Hepatology 1998; 27: 537–545.

    Article  CAS  Google Scholar 

  22. Hofmann AF . Overview of Bile Secretion. In: Pollock DM (ed) Comprehensive Physiology. John Wiley & Sons: Hoboken NJ, 2011, pp 549–566.

    Google Scholar 

  23. Lehmann GL, Carreras FI, Soria LR, Gradilone SA, Marinelli RA . LPS induces the TNF-alpha-mediated downregulation of rat liver aquaporin-8: role in sepsis-associated cholestasis. Am J Physiol Gastrointest Liver Physiol 2008; 294: G567–G575.

    Article  CAS  Google Scholar 

  24. Reinhart WH, Näf G, Werth B . Viscosity of human bile sampled from the common bile duct. Clin Hemorheol Microcirc 2010; 44: 177–182.

    PubMed  Google Scholar 

  25. Roma MG, Toledo FD, Boaglio AC, Basiglio CL, Crocenzi FA, Sánchez Pozzi EJ . Ursodeoxycholic acid in cholestasis: linking action mechanisms to therapeutic applications. Clin Sci (Lond) 2011; 121: 523–544.

    Article  CAS  Google Scholar 

  26. Jarabak J, Talalay P . Stereospecificity of hydrogen transfer by pyridine nucleotide-linked hydroxysteroid dehydrogenases. J Biol Chem 1960; 235: 2147–2151.

    CAS  PubMed  Google Scholar 

  27. Tietz PS, Thistle JL, Miller LJ, LaRusso NF . Development and validation of a method for measuring the glycine and taurine conjugates of bile acids in bile by high-performance liquid chromatography. J Chromatogr 1984; 336: 249–257.

    Article  CAS  Google Scholar 

  28. Carreras FI, Gradilone SA, Mazzone A, García F, Huang BQ, Ochoa JE et al. Rat hepatocyte aquaporin-8 water channels are down-regulated in extrahepatic cholestasis. Hepatology 2003; 37: 1026–1033.

    Article  CAS  Google Scholar 

  29. Marinelli RA, Tietz PS, Caride AJ, Huang BQ, LaRusso NF . Water transporting properties of hepatocyte basolateral and canalicular plasma membrane domains. J Biol Chem 2003; 278: 43157–43162.

    Article  CAS  Google Scholar 

  30. Lowry OH, Rosebrough NJ, Farr AL, Randal RJJ . Protein measurement with the Folin phenol reagent. J Biol Chem 1951; 193: 265–275.

    CAS  Google Scholar 

  31. Rasband, WS. US National Institutes of Health, Bethesda, MD, USA, 1997–2011

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We thank R Rassia for helping us in stopped-flow studies, J Monti for surgical assistance and A Martínez for immunohistochemistry assays. This work was supported by Grant PIP 0244 from Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET) to RAM.

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Correspondence to R A Marinelli.

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Marrone, J., Lehmann, G., Soria, L. et al. Adenoviral transfer of human aquaporin -1 gene to rat liver improves bile flow in estrogen-induced cholestasis. Gene Ther 21, 1058–1064 (2014).

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