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Bile duct ligation causes opposite impacts on the expression and function of BCRP and P-gp in rat brain partly via affecting membrane expression of ezrin/radixin/moesin proteins

Abstract

Breast cancer resistance protein (BCRP) and P-glycoprotein (P-gp) are co-located at blood–brain barrier (BBB) cells, preventing their substrates from entering brain. Accumulating evidence demonstrates that liver failure impairs P-gp and BCRP expression and function in the brain. In the current study, we investigated how liver failure influenced the expression and function of brain BCRP and P-gp in rats subjected to bile duct ligation (BDL). The function of BCRP, P-gp and BBB integrity was assessed using distribution of prazosin, rhodamine 123 and fluorescein, respectively. We showed that BDL significantly decreased BCRP function, but increased P-gp function without affecting BBB integrity. Furthermore, we found that BDL significantly downregulated the expression of membrane BCRP and upregulated the expression of membrane P-gp protein in the cortex and hippocampus. In human cerebral microvascular endothelial cells, NH4Cl plus unconjugated bilirubin significantly decreased BCRP function and expression of membrane BCRP protein, but upregulated P-gp function and expression of membrane P-gp protein. The decreased expression of membrane BCRP protein was linked to the decreased expression of membrane radixin protein, while the increased expression of membrane P-gp protein was related to the increased location of membrane ezrin protein. Silencing ezrin impaired membrane location of P-gp, whereas silencing radixin impaired membrane location of BCRP protein. BDL rats showed the increased expression of membrane ezrin protein and decreased expression of membrane radixin protein in the brain. We conclude that BDL causes opposite effects on the expression and function of brain BCRP and P-gp, attributing to the altered expression of membrane radixin and ezrin protein, respectively, due to hyperbilirubinemia and hyperammonemia.

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Fig. 1: Effects of BDL on the function and expression of BCRP and P-gp in rat brains.
Fig. 2: Effects of NH4Cl, UCB and NH4Cl plus UCB on the function and expression of BCRP and P-gp in HCMEC/D3 cells.
Fig. 3: Effects of BDL on the total and membrane expression of the ERM proteins in rat brains.
Fig. 4: Effects of NH4Cl, UCB and NH4Cl plus UCB on the total and membrane expression of the ERM proteins in HCMEC/D3 cells.
Fig. 5: Effects of silencing ezrin, radixin and moesin on the membrane expression of the BCRP and P-gp proteins.

References

  1. 1.

    Wijdicks EF. Hepatic encephalopathy. N. Engl J Med 2016;315:1660–70.

    Article  CAS  Google Scholar 

  2. 2.

    Aldridge DR, Tranah EJ, Shawcross DL. Pathogenesis of hepatic encephalopathy: role of ammonia and systemic inflammation. J Clin Exp Hepatol. 2015;5:S7–S20.

    PubMed  Article  PubMed Central  Google Scholar 

  3. 3.

    Gil-Martins E, Barbosa DJ, Silva V, Remião F, Silva R. Dysfunction of ABC transporters at the blood-brain barrier: Role in neurological disorders. Pharmacol Ther. 2020;213:107554.

    CAS  PubMed  Article  PubMed Central  Google Scholar 

  4. 4.

    Sun H, Dai H, Shaik N, Elmquist WF. Drug efflux transporters in the CNS. Adv Drug Deliv Rev. 2003;55:83–105.

    CAS  PubMed  Article  PubMed Central  Google Scholar 

  5. 5.

    Ding Y, Wang R, Zhang J, Zhao A, Lu H, Li W, et al. Potential regulation mechanisms of P-gp in the blood-brain barrier in hypoxia. Curr Pharm Des. 2019;25:1041–51.

    CAS  PubMed  Article  Google Scholar 

  6. 6.

    Cui Y, Lotz R, Rapp H, Klinder K, Himstedt A, Sauer A. Muscle to brain partitioning as measure of transporter-mediated efflux at the rat blood-brain barrier and its implementation into compound optimization in drug discovery. Pharmaceutics. 2019;11:595.

    CAS  PubMed Central  Article  Google Scholar 

  7. 7.

    Liu L, Liu X. Contributions of drug transporters to blood-brain barriers. Adv Exp Med Biol. 2019;1141:407–66.

    CAS  PubMed  Article  Google Scholar 

  8. 8.

    Xu P, Ling ZL, Zhang J, Li Y, Shu N, Zhong ZY, et al. Unconjugated bilirubin elevation impairs the function and expression of breast cancer resistance protein (BCRP) at the blood-brain barrier in bile duct-ligated rats. Acta Pharmacol Sin. 2016;37:1129–40.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  9. 9.

    Jin S, Wang XT, Liu L, Yao D, Liu C, Zhang M, et al. P-glycoprotein and multidrug resistance-associated protein 2 are oppositely altered in brain of rats with thioacetamide-induced acute liver failure. Liver Int. 2013;33:274–82.

    CAS  PubMed  Article  Google Scholar 

  10. 10.

    Li Y, Zhang J, Xu P, Sun B, Zhong Z, Liu C, et al. Acute liver failure impairs function and expression of breast cancer-resistant protein (BCRP) at rat blood-brain barrier partly via ammonia-ROS-ERK1/2 activation. J Neurochem. 2016;138:282–94.

    CAS  PubMed  Article  Google Scholar 

  11. 11.

    Liu L, Miao M, Chen Y, Wang Z, Sun B, Liu X, et al. The attenuated function and expression of P-glycoprotein at blood brain barrier of mice by acute liver failure and its clinical significance. Front Pharmacol. 2018;9:190.

    PubMed  PubMed Central  Article  CAS  Google Scholar 

  12. 12.

    Fan Y, Liu X. Alterations in expression and function of ABC family transporters at blood-brain barrier under liver failure and their clinical significances. Pharmaceutics. 2018;10:102.

    CAS  PubMed Central  Article  Google Scholar 

  13. 13.

    Zwingmann C, Butterworth R. An update on the role of brain glutamine synthesis and its relation to cell-specific energy metabolism in the hyperammonemic brain: further studies using NMR spectroscopy. Neurochem Int. 2005;47:19–30.

    CAS  PubMed  Article  PubMed Central  Google Scholar 

  14. 14.

    Kerbert AJC, Jalan R. Recent advances in understanding and managing hepatic encephalopathy in chronic liver disease. F1000Res. 2020;9:F1000 Faculty Rev–312.

  15. 15.

    Gerth HU, Pohlen M, Pavenstädt H, Schmidt H. Extracorporeal liver support of liver failure. Z Gastroenterol. 2017;55:383–93.

    PubMed  Article  Google Scholar 

  16. 16.

    Garcea G, Maddern GJ. Liver failure after major hepatic resection. J Hepatobiliary Pancreat Surg. 2009;16:145–55.

    PubMed  Article  Google Scholar 

  17. 17.

    Zhang J, Zhang M, Sun B, Li Y, Xu P, Liu C, et al. Hyperammonemia enhances the function and expression of P-glycoprotein and Mrp2 at the blood-brain barrier through NF-κB. J Neurochem. 2014;131:791–802.

    CAS  PubMed  Article  Google Scholar 

  18. 18.

    Zhou Y, Zhou J, Li P, Xie Q, Sun B, Li Y, et al. Increase in P-glycoprotein levels in the blood-brain barrier of partial portal vein ligation /chronic hyperammonemia rats is medicated by ammonia/reactive oxygen species/ERK1/2 activation: In vitro and in vivo studies. Eur J Pharmacol. 2019;846:119–27.

    CAS  PubMed  Article  Google Scholar 

  19. 19.

    Kano T, Wada S, Morimoto K, Kato Y, Ogihara T. Effect of knockdown of ezrin, radixin, and moesin on P-glycoprotein function in HepG2 cells. J Pharm Sci. 2011;100:5308–14.

    CAS  PubMed  Article  Google Scholar 

  20. 20.

    Yano K, Okabe C, Fujii K, Kato Y, Ogihara T. Regulation of breast cancer resistance protein and P-glycoprotein by ezrin, radixin and moesin in lung, intestinal and renal cancer cell lines. J Pharm Pharmacol. 2020;72:575–82.

    CAS  PubMed  Article  Google Scholar 

  21. 21.

    Yano K, Otsuka K, Kato Y, Kawabata H, Ohmori S, Arakawa H, et al. Different regulation of P-glycoprotein function between Caco-2 and Caki-1 cells by ezrin, radixin and moesin proteins. J Pharm Pharmacol. 2016;68:361–7.

    CAS  PubMed  Article  PubMed Central  Google Scholar 

  22. 22.

    Pokharel D, Padula MP, Lu JF, Jaiswal R, Djordjevic SP, Bebawy M. The role of CD44 and ERM proteins in expression and functionality of P-glycoprotein in breast cancer cells. Molecules. 2016;21:290.

    PubMed  PubMed Central  Article  CAS  Google Scholar 

  23. 23.

    Hoshi Y, Uchida Y, Kuroda T, Tachikawa M, Couraud PO, Suzuki T, et al. Distinct roles of ezrin, radixin and moesin in maintaining the plasma membrane localizations and functions of human blood-brain barrier transporters. J Cereb Blood Flow Metab. 2020;40:1533–45.

    CAS  PubMed  Article  PubMed Central  Google Scholar 

  24. 24.

    Qin YY, Xu P, Wu T, Qian CQ, Fan YL, Gen DH, et al. Bile duct ligation enhances AZT CNS toxicity partly by impairing the expression and function of BCRP in rat brain. Acta Pharmacol Sin. 2020;41:181–91.

    CAS  PubMed  Article  PubMed Central  Google Scholar 

  25. 25.

    Shao M, Peng ZX, Shi CY, Tang R, Manzo LM, Liu Y. A convenient method for hTfR1 inclusion body purification. Prep Biochem Biotechnol. 2015;45:743–53.

    CAS  PubMed  Article  PubMed Central  Google Scholar 

  26. 26.

    Wilner SE, Wengerter B, Maier K, de Lourdes BorbaMagalhães M, Del Amo DS, Pai S, et al. An RNA alternative to human transferrin: a new tool for targeting human cells. Mol Ther Nucleic Acids. 2012;1:e21.

    PubMed  PubMed Central  Article  CAS  Google Scholar 

  27. 27.

    Li S, Wang X, Ma QH, Yang WL, Zhang XG, Dawe GS. Amyloid precursor protein modulates Nav1.6 sodium channel currents through a Go-coupled JNK pathway. Sci Rep. 2016;6:39320.

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  28. 28.

    Abbott NJ, Patabendige AA, Dolman DE, Yusof SR, Begley DJ. Structure and function of the blood-brain barrier. Neurobiol Dis. 2010;37:13–25.

    CAS  PubMed  Article  PubMed Central  Google Scholar 

  29. 29.

    Begley D. ABC transporters and the blood-brain barrier. Curr Pharm Des. 2004;10:1295–312.

    CAS  PubMed  Article  Google Scholar 

  30. 30.

    Scherrmann J. Expression and function of multidrug resistance transporters at the blood-brain barriers. Expert Opin Drug Metab Toxicol. 2005;1:233–46.

    CAS  PubMed  Article  PubMed Central  Google Scholar 

  31. 31.

    Liu XD. ABC family transporters. AdvExp Med Biol. 2019;1141:13–100.

    CAS  Article  Google Scholar 

  32. 32.

    Amawi H, Sim HM, Tiwari AK, Ambudkar SV, Shukla S. ABC transporter-mediated multidrug-resistant cancer. Adv Exp Med Biol. 2019;1141:549–80.

    CAS  PubMed  Article  Google Scholar 

  33. 33.

    Huang HC, Chang CC, Wang SS, Chan CY, LeeFT, Chuang CL, et al. Pravastatin for thioacetamide-induced hepatic failure and encephalopathy. Eur J Clin Invest. 2012;42:139–45.

    CAS  PubMed  Article  Google Scholar 

  34. 34.

    Mendes NF, Mariotti FFN, de Andrade JS, de Barros Viana M, Céspedes IC, Nagaoka MR, et al. Lactulose decreases neuronal activation and attenuates motor behavioral deficits in hyperammonemic rats. Metab Brain Dis. 2017;32:2073–83.

    CAS  PubMed  Article  Google Scholar 

  35. 35.

    Radeva G, Perabo J, Sharom FJ. P-glycoprotein is localized in intermediate-density membrane microdomains distinct from classical lipid rafts and caveolar domains. FEBS J. 2005;272:4924–37.

    CAS  PubMed  Article  Google Scholar 

  36. 36.

    Storch CH, EhehaltR, Haefeli WE, Weiss J. Localization of the human breast cancer resistance protein (BCRP/ABCG2) in lipid rafts/caveolae and modulation of its activity by cholesterol in vitro. J Pharmacol Exp Ther. 2007;323:257–64.

    CAS  PubMed  Article  Google Scholar 

  37. 37.

    Gayet L, Dayan G, Barakat S, Labialle S, Michaud M, Cogne S, et al. Control of P-glycoprotein activity by membrane cholesterol amounts and their relation to multidrug resistance in human CEM leukemia cells. Biochemistry. 2005;44:4499–509.

    CAS  PubMed  Article  PubMed Central  Google Scholar 

  38. 38.

    Tanaka S, Kinowaki M, Maeda Y, Nagatomo J, Kai MH, Kondo KH, et al. Speciesdifference incholesterol7alpha-hydroxylase expression of rabbit and rat liver microsomes after bile duct ligation. J Surg Res. 2004;119:36–40.

    CAS  PubMed  Article  PubMed Central  Google Scholar 

  39. 39.

    Kobori T, Harada S, Nakamoto K, Tokuyama S. Radixin influences the changes in the small intestinal P-glycoprotein by etoposide treatment. Biol Pharm Bull. 2013;36:1822–8.

    CAS  PubMed  Article  PubMed Central  Google Scholar 

  40. 40.

    Yano K, Tomono T, Sakai R, Kano T, Morimoto K, Kato Y, et al. Contribution of radixin to P-glycoprotein expression and transport activity in mouse small intestine in vivo. J Pharm Sci. 2013;102:2875–81.

    CAS  PubMed  Article  PubMed Central  Google Scholar 

  41. 41.

    Kobori T, Fujiwara S, Miyagi K, Harada S, Nakamoto K, Nakagawa T, et al. Involvement of moesin in the development of morphine analgesic tolerance through P-glycoprotein at the blood-brain barrier. Drug Metab Pharmacokinet. 2014;29:482–9.

    PubMed  Article  CAS  Google Scholar 

  42. 42.

    Zhang Y, Dong J, Zhu X, Wang W, Yang Q. The effect of sphingomyelin synthase 2 (SMS2) deficiency on the expression of drug transporters in mouse brain. Biochem Pharmacol. 2011;82:287–94.

    CAS  PubMed  Article  PubMed Central  Google Scholar 

  43. 43.

    Shawcross DL, Wendon A. The neurological manifestations of acute liver failure. Neurochem Inter. 2012;60:662–71.

    CAS  Article  Google Scholar 

  44. 44.

    De Souza J, Benet LZ, Huang Y, Storpirtis S. Comparison of bidirectional lamivudine and zidovudine transport using MDCK, MDCK-MDR1, and Caco-2 cell monolayers. J Pharm Sci. 2009;98:4413–9.

    PubMed  Article  CAS  Google Scholar 

  45. 45.

    Shaik N, Giri N, Pan G, Elmquist WF. P-glycoprotein-mediated active efflux of the anti-HIV1 nucleoside abacavir limits cellular accumulation and brain distribution. Drug Metab Dispos. 2007;35:2076–85.

    CAS  PubMed  Article  Google Scholar 

  46. 46.

    Pan G, Giri N, Elmquist WF. Abcg2/Bcrp1 mediates the polarized transport of antiretroviral nucleosides abacavir and zidovudine. Drug Metab Dispos. 2007;35:1165–73.

    CAS  PubMed  Article  Google Scholar 

  47. 47.

    Römermann K, Helmer R, Löscher W. The antiepileptic druglamotrigineis a substrate of mouse and human breast cancer resistance protein (ABCG2). Neuropharmacology. 2015;93:7–14.

    PubMed  Article  CAS  PubMed Central  Google Scholar 

  48. 48.

    Yang ZH, Liu XD. P-glycoprotein-mediated efflux of phenobarbital at the blood-brain barrier evidence from transport experiments in vitro. Epilepsy Res. 2008;78:40–9.

    CAS  PubMed  Article  Google Scholar 

  49. 49.

    Clucas J, Valderrama F. ERM proteins in cancer progression. J Cell Sci. 2014;127:267–75.

    CAS  PubMed  Article  Google Scholar 

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Acknowledgements

This study was supported by the National Natural Science Foundation of China (Nos. 81872930, 82073922, 81673505), and the “Double First-Class” University Project (CPU2018GY22).

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TW, YYQ, XDL, and LL designed the experiments. TW and XDL analyzed the data. TW, XDL and LL wrote the manuscript. TW, YS, WMK, MMJ, HYY, XKZ, CD, ML conducted experiments. TW, YS, XDL and LL reviewed the manuscript.

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Correspondence to Xiao-dong Liu or Li Liu.

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Wu, T., Sheng, Y., Qin, Yy. et al. Bile duct ligation causes opposite impacts on the expression and function of BCRP and P-gp in rat brain partly via affecting membrane expression of ezrin/radixin/moesin proteins. Acta Pharmacol Sin (2021). https://doi.org/10.1038/s41401-020-00602-3

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Keywords

  • liver failure
  • breast cancer resistance protein
  • P-glycoprotein
  • blood-brain barrier
  • ERM proteins

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