Time-dependent modulation of FXR and Nrf2 signaling in rat models of BDL-induced cholestasis

Articles in Press

Document Type : Original Article

Authors

1 Evidence-based Phytotherapy and Complementary Medicine Research Center, Alborz University of Medical Sciences, Karaj, Iran

2 Cardiovascular Research Center, Alborz University of Medical Sciences, Karaj, Iran

3 Department of Physiology-Pharmacology-Medical Physics, School of Medicine, Alborz University of Medical Sciences, Karaj, Iran

4 Department of Pathology, School of Medicine, Alborz University of Medical Sciences, Karaj, Iran

5 Department of Cardiology, School of Medicine, Alborz University of Medical Sciences, Karaj, Iran

10.22038/ijbms.2026.90035.19420

Abstract

Objective(s): Farnesoid X receptor (FXR) and nuclear factor erythroid 2-related factor 2 (Nrf2) protect the liver against cholestatic injury by regulating antioxidant and anti-inflammatory pathways. Bile duct ligation (BDL) is a standard model for studying cholestatic liver disease, yet the temporal dynamics of FXR/Nrf2 signaling and their downstream mediators remain unclear. To investigate time-dependent changes in hepatic FXR, Nrf2, and downstream effectors over six weeks following BDL in rats, to identify potential preventive and therapeutic targets.
Materials and Methods: Forty-nine male Wistar rats were divided into one sham-operated group and six BDL groups, sacrificed sequentially from weeks 1 to 6 post-surgery. Biochemical assays, histopathology, and molecular analyses were performed to assess dynamic changes in FXR, Nrf2, and related oxidative stress and inflammatory markers.
Results: The most severe histological distortions were observed mainly at week six. Expression reductions in FXR, Superoxide Dismutase (SOD), Alpha-Glutathione S-Transferase (α-GST), and Glutamate-Cysteine Ligase Modifier Subunit (GCLM) were notable from week 1 to week 6, significantly declining in BDL rats compared to sham-operated ones. Nrf2 notably decreased at week 4 post-BDL (P-value<0.05). TNF-α exhibited an increasing trend, peaking at week 6 (P<0.001). FXR prominently decreased in weeks 1 and 5 (P<0.001), and SOD notably decreased in week 5 (P<0.05). α-GST and GCLM also markedly decreased, especially during BDL-W3 and BDL-W4 (P<0.001).
Conclusion: Temporal suppression of FXR/Nrf2 signaling and antioxidant defenses likely contribute to BDL-induced cholestatic injury, with a biphasic pattern across acute and sub-acute phases. These pathways may serve as therapeutic targets during disease progression.

Keywords

Main Subjects

References

1. Hirschfield GM, Heathcote EJ, Gershwin ME. Pathogenesis of cholestatic liver disease and therapeutic approaches. Gastroenterology 2010; 139: 1481-1496. 
2. Cai SY, Boyer JL. The role of bile acids in cholestatic liver injury. Ann Transl Med 2021; 9: 608. 
3. Li M, Cai SY, Boyer JL. Mechanisms of bile acid mediated inflammation in the liver. Mol Aspects Med 2017; 56: 45–53. 
4. Schuppan D, Afdhal NH. Liver cirrhosis. Lancet 2008; 371: 838-851. 
5. Petrov PD, Soluyanova P, Sánchez-Campos S, Castell JV, Jover R. Molecular mechanisms of hepatotoxic cholestasis by clavulanic acid: Role of NRF2 and FXR pathways. Food Chem Toxicol 2021; 158: 112664. 
6. Petrescu AD, DeMorrow S. Farnesoid X receptor as target for therapies to treat cholestasis-induced liver injury. Cells 2021; 10: 1846. 
7. Panzitt K, Zollner G, Marschall HU, Wagner M. Recent advances on FXR-targeting therapeutics. Mol Cell Endocrinol 2022; 552: 111678. 
8. Yang H, Ramani K, Xia M, Ko KS, Li TW, Oh P, et al. Dysregulation of glutathione synthesis during cholestasis in mice: Molecular mechanisms and therapeutic implications. Hepatology 2009; 49: 1982-1991. 
9. Mathan Kumar S, Dey A. Regulation of glutathione in health and disease with special emphasis on chronic alcoholism and hyperglycaemia mediated liver injury: A brief perspective. Springer Sci Rev 2014; 2: 1–13. 
10. Senoner T, Schindler S, Stättner S, Öfner D, Troppmair J, Primavesi F. Associations of oxidative stress and postoperative outcome in liver surgery with an outlook to future potential therapeutic options. Oxid Med Cell Longev 2019; 2019: 3950818. 
11. Galicia-Moreno M, Lucano-Landeros S, Monroy-Ramirez HC, Silva-Gomez J, Gutierrez-Cuevas J, Santos A, et al. Roles of Nrf2 in liver diseases: Molecular, pharmacological, and epigenetic aspects. Antioxidants 2020; 9: 980. 
12. Stofan M, Guo GL. Bile acids and FXR: Novel targets for liver diseases. Front Med 2020; 7: 544. 
13. Verbeke L, Mannaerts I, Schierwagen R, Govaere O, Klein S, Vander Elst I, et al. FXR agonist obeticholic acid reduces hepatic inflammation and fibrosis in a rat model of toxic cirrhosis. Sci Rep 2016; 6: 33453. 
14. Yang Y, Chen B, Chen Y, Zu B, Yi B, Lu K. A comparison of two common bile duct ligation methods to establish hepatopulmonary syndrome animal models. Lab Anim 2015; 49: 71–79. 
15. Halilbasic E, Baghdasaryan A, Trauner M. Nuclear receptors as drug targets in cholestatic liver diseases. Clin Liver Dis 2013; 17: 161-189. 
16. Zollner G, Wagner M, Fickert P, Silbert D, Gumhold J, Zatloukal K, et al. Expression of bile acid synthesis and detoxification enzymes and the alternative bile acid efflux pump MRP4 in patients with primary biliary cirrhosis. Liver Int 2007; 27: 920–929. 
17. Demeilliers C, Jacquemin E, Barbu V, Mergey M, Paye F, Fouassier L, et al. Altered hepatobiliary gene expressions in PFIC1: ATP8B1 gene defect is associated with CFTR downregulation. Hepatology 2006; 43: 1125–1134. 
18. Alvarez L, Jara P, Sánchez-Sabate E, Hierro L, Larrauri J, Díaz MC, et al. Reduced hepatic expression of farnesoid X receptor in hereditary cholestasis associated to mutation in ATP8B1. Hum Mol Genet 2004; 13: 2451–2560. 
19. Gai Z, Chu L, Xu Z, Song X, Sun D, Kullak-Ublick GA. Farnesoid X receptor activation protects the kidney from ischemia-reperfusion damage. Sci Rep 2017; 7: 9815. 
20. Bayat G, Hashemi SA, Karim H, Fallah P, Hedayatyanfard K, Bayat M, et al. Biliary cirrhosis-induced cardiac abnormality in rats: Interaction between Farnesoid-X-activated receptors and the cardiac uncoupling proteins 2 and 3. Iran J Basic Med Sci 2022; 25: 126-133. 
21. Wagner M, Fickert P, Zollner G, Fuchsbichler A, Silbert D, Tsybrovskyy O, et al. Role of farnesoid X receptor in determining hepatic ABC transporter expression and liver injury in bile duct-ligated mice. Gastroenterology 2003; 125: 825–838. 
22. Maher J, Yamamoto M. The rise of antioxidant signaling—the evolution and hormetic actions of Nrf2. Toxicol Appl Pharmacol 2010; 244: 4–15. 
23. Xu Z, Tang W, Xie Q, Cao X, Zhang M, Zhang X, et al. Dimethyl fumarate attenuates cholestatic liver injury by activating the NRF2 and FXR pathways and suppressing NLRP3/GSDMD signaling in mice. Exp Cell Res 2023; 432: 113781. 
24. Yang D, Chen X, Wang J, Lou Q, Lou Y, Li L, et al. Dysregulated lung commensal bacteria drive interleukin-17B production to promote pulmonary fibrosis through their outer membrane vesicles. Immunity 2019; 50: 692-706. 
25. He L, Guo C, Peng C, Li Y. Advances of natural activators for Nrf2 signaling pathway on cholestatic liver injury protection: A review. Eur J Pharmacol 2021; 910: 174447. 
26. Fahmy SR. Anti-fibrotic effect of Holothuria arenicola extract against bile duct ligation in rats. BMC Complement Altern Med 2015; 15: 1-12. 
27. Sánchez-Valle V, C Chavez-Tapia N, Uribe M, Méndez-Sánchez N. Role of oxidative stress and molecular changes in liver fibrosis: A review. Curr Med Chem 2012; 19: 4850–4860. 
28. Nguyen T, Nioi P, Pickett CB. The Nrf2-antioxidant response element signaling pathway and its activation by oxidative stress. J Biol Chem 2009; 284: 13291–13295. 
29. Mohs A, Otto T, Schneider KM, Peltzer M, Boekschoten M, Holland CH, et al. Hepatocyte-specific NRF2 activation controls fibrogenesis and carcinogenesis in steatohepatitis. J Hepatol 2021; 74: 638–648. 
30. Fuertes-Agudo M, Luque-Tévar M, Cucarella C, Martín-Sanz P, Casado M. Advances in understanding the role of NRF2 in liver pathophysiology and its relationship with hepatic-specific cyclooxygenase-2 expression. Antioxidants 2023; 12: 1491. 
31. Seedorf K, Weber C, Vinson C, Berger S, Vuillard LM, Kiss A, et al. Selective disruption of NRF2-KEAP1 interaction leads to NASH resolution and reduction of liver fibrosis in mice. JHEP Rep 2023; 5: 100651. 
32. Wen YA, Liu D, Zhou QY, Huang SF, Luo P, Xiang Y, et al. Biliary intervention aggravates cholestatic liver injury, and induces hepatic inflammation, proliferation and fibrogenesis in BDL mice. Exp Toxicol Pathol 2011; 63: 277–284. 
33. Osawa Y, Hoshi M, Yasuda I, Saibara T, Moriwaki H, Kozawa O. Tumor necrosis factor-α promotes cholestasis-induced liver fibrosis in the mouse through tissue inhibitor of metalloproteinase-1 production in hepatic stellate cells. PloS One 2013; 8: e65251. 
34. Gäbele E, Froh M, Arteel GE, Uesugi T, Hellerbrand C, Schölmerich J, et al. TNFα is required for cholestasis-induced liver fibrosis in the mouse. Biochem Biophys Res Commun 2009; 378: 348–353. 
35. Lopetuso LR, Mocci G, Marzo M, D’Aversa F, Rapaccini GL, Guidi L, et al. Harmful effects and potential benefits of anti-tumor necrosis factor (TNF)-α on the liver. Int J Mol Sci 2018; 19: 2199.  
Iranian Journal of Basic Medical Sciences

Articles in Press, Accepted Manuscript
Available Online from 17 May 2026

Files

Share

How to cite

Statistics