Articles
Vol. 70 No. 4 (2026)
Discovering ferroptosis in biliary epithelium during liver fibrosis
Publisher's note
All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.
All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.
Published: 28 October 2026
39
Views
21
Downloads
Authors
Ferroptosis is an iron-dependent regulated cell death characterized by iron accumulation and lipid peroxidation. The liver represents the primary site of iron-overload injury due to its role in iron metabolism, but the potential role of ferroptosis in the biliary epithelium is yet unknown. The study investigated the recruitment of ferroptosis under control conditions and after bile duct ligation (BDL), highlighting the effects of ferroptosis inducers and inhibitors in a cell line of cholangiocytes. In addition, we identified iron deposits through Perls Prussian blue as well as the presence of proteins such as ferritin, transferrin receptor 1 (CD71), glutathione peroxidase-4 (GPX4) and acyl-CoA synthetase long-chain family member 4 (ACSL4). The results showed an increase in the expression of ferritin, CD71 and ACSL4 in the BDL model, whereas GPX4 was reduced in the cholestatic model in comparison to the control. In addition, sorafenib failed to trigger ferroptosis in cholangiocytes, whereas erastin was a potent inducer. Conversely, ferrostatin-1 (Fer-1) did not inhibit ferroptosis, while β-mercaptoethanol (βME) could protect cholangiocytes from ferroptosis-induced cell death. All these findings are important for understanding the involvement of ferroptosis in cholangiocyte pathophysiology and may support additional studies regarding the possible application of ferroptosis modulators in the treatment of liver fibrosis.
Downloads
Download data is not yet available.
Citations
1. Galluzzi L, Bravo-San Pedro JM, Kepp O, Kroemer G. Regulated cell death and adaptive stress responses. Cell Mol Life Sci 2016;73:2405-10. DOI: https://doi.org/10.1007/s00018-016-2209-y
2. Galluzzi L, Vitale I, Aaronson SA, Abrams JM, Adam D, Agostinis P, et al. Molecular mechanisms of cell death: recommendations of the Nomenclature Committee on Cell Death 2018. Cell Death Differ 2018;25:486-541. DOI: https://doi.org/10.1038/s41418-018-0102-y
3. Galluzzi L, Vitale I, Abrams JM, Alnemri ES, Baehrecke EH, Blagosklonny MV, et al. Molecular definitions of cell death subroutines: recommendations of the Nomenclature Committee on Cell Death 2012. Cell Death Differ 2012;19:107-20. DOI: https://doi.org/10.1038/cdd.2011.96
4. Kroemer G, Galluzzi L, Vandenabeele P, Abrams J, Alnemri ES, Baehrecke EH, et al. Classification of cell death: recommendations of the Nomenclature Committee on Cell Death 2009. Cell Death Differ 2009;16:3-11. DOI: https://doi.org/10.1038/cdd.2008.150
5. Dixon SJ, Lemberg KM, Lamprecht MR, Skouta R, Zaitsev EM, Gleason CE, et al. Ferroptosis: an iron-dependent form of nonapoptotic cell death. Cell 2012;149:1060-72. DOI: https://doi.org/10.1016/j.cell.2012.03.042
6. Yang WS, SriRamaratnam R, Welsch ME, Shimada K, Skouta R, Viswanathan VS, et al. Regulation of ferroptotic cancer cell death by GPX4. Cell 2014;156:317-31. DOI: https://doi.org/10.1016/j.cell.2013.12.010
7. Shah R, Margison K, Pratt DA. The potency of diarylamine radical-trapping antioxidants as inhibitors of ferroptosis underscores the role of autoxidation in the mechanism of cell death. ACS Chem Biol 2017;12:2538-45. DOI: https://doi.org/10.1021/acschembio.7b00730
8. Vanden Berghe T, Linkermann A, Jouan-Lanhouet S, Walczak H, Vandenabeele P. Regulated necrosis: the expanding network of non-apoptotic cell death pathways. Nat Rev Mol Cell Biol 2014;15:135-47. DOI: https://doi.org/10.1038/nrm3737
9. Munshi MK, Priester S, Gaudio E, Yang F, Alpini G, Mancinelli R, et al. Regulation of biliary proliferation by neuroendocrine factors: implications for the pathogenesis of cholestatic liver diseases. Am J Pathol 2011;178:472-84. DOI: https://doi.org/10.1016/j.ajpath.2010.09.043
10. Ceci L, Gaudio E, Kennedy L. Cellular Interactions and crosstalk facilitating biliary fibrosis in cholestasis. Cell Mol Gastroenterol Hepatol 2024;17:553-65. DOI: https://doi.org/10.1016/j.jcmgh.2024.01.005
11. Wang CY, Babitt JL. Liver iron sensing and body iron homeostasis. Blood 2019;133:18-29. DOI: https://doi.org/10.1182/blood-2018-06-815894
12. Guo J, Duan L, He X, Li S, Wu Y, Xiang G, et al. A combined model of human iPSC-derived liver organoids and hepatocytes reveals ferroptosis in DGUOK mutant mtDNA depletion syndrome. Adv Sci (Weinh) 2021;8:2004680. DOI: https://doi.org/10.1002/advs.202004680
13. Ali N, Ferrao K, Mehta KJ. Liver iron loading in alcohol-associated liver disease. Am J Pathol 2023;193:1427-39. DOI: https://doi.org/10.1016/j.ajpath.2022.08.010
14. Lu Y, Hu J, Chen L, Li S, Yuan M, Tian X, et al. Ferroptosis as an emerging therapeutic target in liver diseases. Front Pharmacol 2023;14:1196287. DOI: https://doi.org/10.3389/fphar.2023.1196287
15. Kazan HH, Urfali-Mamatoglu C, Gunduz U. Iron metabolism and drug resistance in cancer. Biometals 2017;30:629-41. DOI: https://doi.org/10.1007/s10534-017-0037-7
16. Uchida M, Kang S, Reichhardt C, Harlen K, Douglas T. The ferritin superfamily: Supramolecular templates for materials synthesis. Biochim Biophys Acta 2010;1800:834-45. DOI: https://doi.org/10.1016/j.bbagen.2009.12.005
17. Friedmann Angeli JP, Schneider M, Proneth B, Tyurina YY, Tyurin VA, Hammond VJ, et al. Inactivation of the ferroptosis regulator Gpx4 triggers acute renal failure in mice. Nat Cell Biol 2014;16:1180-91. DOI: https://doi.org/10.1038/ncb3064
18. Tang Y, Zhou J, Hooi SC, Jiang YM, Lu GD. Fatty acid activation in carcinogenesis and cancer development: Essential roles of long-chain acyl-CoA synthetases. Oncol Lett 2018;16:1390-6. DOI: https://doi.org/10.3892/ol.2018.8843
19. Quan J, Bode AM, Luo X. ACSL family: The regulatory mechanisms and therapeutic implications in cancer. Eur J Pharmacol 2021;909:174397. DOI: https://doi.org/10.1016/j.ejphar.2021.174397
20. Nie Q, Hu Y, Yu X, Li X, Fang X. Induction and application of ferroptosis in cancer therapy. Cancer Cell Int 2022;22:12. DOI: https://doi.org/10.1186/s12935-021-02366-0
21. Li Y, Zeng X, Lu D, Yin M, Shan M, Gao Y. Erastin induces ferroptosis via ferroportin-mediated iron accumulation in endometriosis. Hum Reprod 2021;36:951-64. DOI: https://doi.org/10.1093/humrep/deaa363
22. Lou JS, Zhao LP, Huang ZH, Chen XY, Xu JT, Tai WC, et al. Ginkgetin derived from Ginkgo biloba leaves enhances the therapeutic effect of cisplatin via ferroptosis-mediated disruption of the Nrf2/HO-1 axis in EGFR wild-type non-small-cell lung cancer. Phytomedicine 2021;80:153370. DOI: https://doi.org/10.1016/j.phymed.2020.153370
23. Dixon SJ, Patel DN, Welsch M, Skouta R, Lee ED, Hayano M, et al. Pharmacological inhibition of cystine-glutamate exchange induces endoplasmic reticulum stress and ferroptosis. Elife 2014;3:e02523. DOI: https://doi.org/10.7554/eLife.02523
24. Zhang H, Tian L, Wang P, Li L, Wang K, Li Y, et al. Ferrostatin-1 mitigates acute lung injury by reducing ferroptosis levels in gas explosions. Tissue Cell 2025;94:102773. DOI: https://doi.org/10.1016/j.tice.2025.102773
25. Yan J, Bao L, Liang H, Zhao L, Liu M, Kong L, et al. A druglike ferrostatin-1 analogue as a ferroptosis inhibitor and photoluminescent indicator. Angew Chem Int Ed Engl 2025;64:e202502195. DOI: https://doi.org/10.1002/anie.202502195
26. Chen X, Li J, Kang R, Klionsky DJ, Tang D. Ferroptosis: machinery and regulation. Autophagy 2021;17:2054-81. DOI: https://doi.org/10.1080/15548627.2020.1810918
27. Zheng J, Zhang W, Ito J, Henkelmann B, Xu C, Mishima E, et al. N-acetyl-l-cysteine averts ferroptosis by fostering glutathione peroxidase 4. Cell Chem Biol 2025;32:767-75 e5. DOI: https://doi.org/10.1016/j.chembiol.2025.04.002
28. Alpini G, Lenzi R, Sarkozi L, Tavoloni N. Biliary physiology in rats with bile ductular cell hyperplasia. Evidence for a secretory function of proliferated bile ductules. J Clin Invest 1988;81:569-78. DOI: https://doi.org/10.1172/JCI113355
29. Hall AP, Davies W, Stamp K, Clamp I, Bigley A. Comparison of computerized image analysis with traditional semiquantitative scoring of Perls' Prussian Blue stained hepatic iron deposition. Toxicol Pathol 2013;41:992-1000. DOI: https://doi.org/10.1177/0192623313476576
30. Shao M, Qi K, Wang L, Yu X, Zhang Q, Yu L, et al. E3 ubiquitin ligase CHIP interacts with transferrin receptor 1 for degradation and promotes cell proliferation through inhibiting ferroptosis in hepatocellular carcinoma. Cell Signal 2024;118:111148. DOI: https://doi.org/10.1016/j.cellsig.2024.111148
31. Guo S, Zhang D, Dong Y, Shu Y, Wu X, Ni Y, et al. Sulfiredoxin-1 accelerates erastin-induced ferroptosis in HT-22 hippocampal neurons by driving heme Oxygenase-1 activation. Free Radic Biol Med 2024;223:430-42. DOI: https://doi.org/10.1016/j.freeradbiomed.2024.08.008
32. Ma W, Jia L, Zhao J, Li Y, Kong J, Du H. Iron regulates lipid droplet formation in hepatocytes via heme oxygenase-1 mediated ferroptosis. J Trace Elem Med Biol 2025;91:127719. DOI: https://doi.org/10.1016/j.jtemb.2025.127719
33. Jiang Y, Yu Y, Pan Z, Glandorff C, Sun M. Ferroptosis: a new hunter of hepatocellular carcinoma. Cell Death Discov 2024;10:136. DOI: https://doi.org/10.1038/s41420-024-01863-1
34. Ajoolabady A, Tang D, Kroemer G, Ren J. Ferroptosis in hepatocellular carcinoma: mechanisms and targeted therapy. Br J Cancer 2023;128:190-205. DOI: https://doi.org/10.1038/s41416-022-01998-x
35. Zhao J, Xu B, Xiong Q, Feng Y, Du H. Erastin induced ferroptosis causes physiological and pathological changes in healthy tissues of mice. Mol Med Rep 2021;24:713. DOI: https://doi.org/10.3892/mmr.2021.12352
36. Keating GM. Sorafenib: a review in hepatocellular carcinoma. Target Oncol 2017;12:243-53. DOI: https://doi.org/10.1007/s11523-017-0484-7
37. Escudier B, Worden F, Kudo M. Sorafenib: key lessons from over 10 years of experience. Expert Rev Anticancer Ther 2019;19:177-89. DOI: https://doi.org/10.1080/14737140.2019.1559058
38. Zheng J, Sato M, Mishima E, Sato H, Proneth B, Conrad M. Sorafenib fails to trigger ferroptosis across a wide range of cancer cell lines. Cell Death Dis 2021;12:698. DOI: https://doi.org/10.1038/s41419-021-03998-w
39. Miotto G, Rossetto M, Di Paolo ML, Orian L, Venerando R, Roveri A, et al. Insight into the mechanism of ferroptosis inhibition by ferrostatin-1. Redox Biol 2020;28:101328. DOI: https://doi.org/10.1016/j.redox.2019.101328
40. Delehouze C, Comte A, Leon-Icaza SA, Cougoule C, Hauteville M, Goekjian P, et al. Nigratine as dual inhibitor of necroptosis and ferroptosis regulated cell death. Sci Rep 2022;12:5118. DOI: https://doi.org/10.1038/s41598-022-09019-w
41. Martinez AM, Kim A, Yang WS. Detection of Ferroptosis by BODIPY 581/591 C11. Methods Mol Biol 2020;2108:125-30. DOI: https://doi.org/10.1007/978-1-0716-0247-8_11
42. Liu Y, Zhang M, Huang Y, Zhang Y, Zhu C, Guan J, et al. Involvement of ferroptosis in metabolic dysfunction-associated steatohepatitis-related liver diseases. J Mol Med (Berl) 2025;103:1299-315. DOI: https://doi.org/10.1007/s00109-025-02599-w
43. Chen J, Li X, Ge C, Min J, Wang F. The multifaceted role of ferroptosis in liver disease. Cell Death Differ 2022;29:467-80. DOI: https://doi.org/10.1038/s41418-022-00941-0
44. Franca M, Marti-Bonmati L, Porto G, Silva S, Guimaraes S, Alberich-Bayarri A, et al. Tissue iron quantification in chronic liver diseases using MRI shows a relationship between iron accumulation in liver, spleen, and bone marrow. Clin Radiol 2018;73:215.e1-9. DOI: https://doi.org/10.1016/j.crad.2017.07.022
45. Theurl I, Hilgendorf I, Nairz M, Tymoszuk P, Haschka D, Asshoff M, et al. On-demand erythrocyte disposal and iron recycling requires transient macrophages in the liver. Nat Med 2016;22:945-51. DOI: https://doi.org/10.1038/nm.4146
46. Tao L, Yang X, Ge C, Zhang P, He W, Xu X, et al. Integrative clinical and preclinical studies identify FerroTerminator1 as a potent therapeutic drug for MASH. Cell Metab 2024;36:2190-206.e5. DOI: https://doi.org/10.1016/j.cmet.2024.07.013
47. Kim HG, Han JM, Lee HW, Lee JS, Son SW, Choi MK, et al. CGX, a multiple herbal drug, improves cholestatic liver fibrosis in a bile duct ligation-induced rat model. J Ethnopharmacol 2013;145:653-62. DOI: https://doi.org/10.1016/j.jep.2012.12.005
48. Liu T, Liu W, Zhang M, Yu W, Gao F, Li C, et al. Ferrous-supply-regeneration nanoengineering for cancer-cell-specific ferroptosis in combination with imaging-guided photodynamic therapy. ACS Nano 2018;12:12181-92. DOI: https://doi.org/10.1021/acsnano.8b05860
49. Arosio P, Levi S. Ferritin, iron homeostasis, and oxidative damage. Free Radic Biol Med 2002;33:457-63. DOI: https://doi.org/10.1016/S0891-5849(02)00842-0
50. Reikvam H, Rolfsnes MG, Rolsdorph L, Sandnes M, Selheim F, Hernandez-Valladares M, et al. Ferritin in acute myeloid leukemia: not only a marker of inflammation and iron overload, but also a regulator of cellular iron metabolism, signaling and communication. Int J Mol Sci 2025;26:5744. DOI: https://doi.org/10.3390/ijms26125744
51. Luo Y, Liu C, Yao Y, Tang X, Yin E, Lu Z, et al. A comprehensive pan-cancer analysis of prognostic value and potential clinical implications of FTH1 in cancer immunotherapy. Cancer Immunol Immunother 2024;73:37. DOI: https://doi.org/10.1007/s00262-023-03625-x
52. Piccolo M, Ferraro MG, Iazzetti F, Santamaria R, Irace C. Insight into iron, oxidative stress and ferroptosis: therapy targets for approaching anticancer strategies. Cancers (Basel) 2024;16:1220. DOI: https://doi.org/10.3390/cancers16061220
53. Wang S, Guo Q, Zhou L, Xia X. Ferroptosis: A double-edged sword. Cell Death Discov 2024;10:265. DOI: https://doi.org/10.1038/s41420-024-02037-9
54. Sinha BK, Murphy C, Brown SM, Silver BB, Tokar EJ, Bortner CD. Mechanisms of cell death induced by erastin in human ovarian tumor cells. Int J Mol Sci 2024;25:8666. DOI: https://doi.org/10.3390/ijms25168666
55. Zhao Y, Li Y, Zhang R, Wang F, Wang T, Jiao Y. The role of erastin in ferroptosis and its prospects in cancer therapy. Onco Targets Ther 2020;13:5429-41. DOI: https://doi.org/10.2147/OTT.S254995
56. Hao SH, Ma XD, Xu L, Xie JD, Feng ZH, Chen JW, et al. Dual specific phosphatase 4 suppresses ferroptosis and enhances sorafenib resistance in hepatocellular carcinoma. Drug Resist Updat 2024;73:101052. DOI: https://doi.org/10.1016/j.drup.2024.101052
57. Liu L, Cao Y, Chen C, Zhang X, McNabola A, Wilkie D, et al. Sorafenib blocks the RAF/MEK/ERK pathway, inhibits tumor angiogenesis, and induces tumor cell apoptosis in hepatocellular carcinoma model PLC/PRF/5. Cancer Res 2006;66:11851-8. DOI: https://doi.org/10.1158/0008-5472.CAN-06-1377
58. Rini BI, Escudier B, Tomczak P, Kaprin A, Szczylik C, Hutson TE, et al. Comparative effectiveness of axitinib versus sorafenib in advanced renal cell carcinoma (AXIS): a randomised phase 3 trial. Lancet 2011;378:1931-9. DOI: https://doi.org/10.1016/S0140-6736(11)61613-9
59. Wang W, Lu K, Jiang X, Wei Q, Zhu L, Wang X, et al. Ferroptosis inducers enhanced cuproptosis induced by copper ionophores in primary liver cancer. J Exp Clin Cancer Res 2023;42:142. DOI: https://doi.org/10.1186/s13046-023-02720-2
60. Li Y, Yang W, Zheng Y, Dai W, Ji J, Wu L, et al. Targeting fatty acid synthase modulates sensitivity of hepatocellular carcinoma to sorafenib via ferroptosis. J Exp Clin Cancer Res 2023;42:6. DOI: https://doi.org/10.1186/s13046-022-02567-z
61. Jiang H, Zhang X, Yang W, Li M, Wang G, Luo Q. Ferrostatin-1 ameliorates liver dysfunction via reducing iron in thioacetamide-induced acute liver injury in mice. Front Pharmacol 2022;13:869794. DOI: https://doi.org/10.3389/fphar.2022.869794
Ethics Approval
The animal studies were performed following the regulations of the protocols approved by Baylor Scott & White Institutional Animal Care and Use CommitteeCRediT authorship contribution
Romina Mancinelli, conceptualization, validation, resources, funding acquisition, writing - original draft. Arianna Casini, methodology, resources, funding acquisition. Stefano Leone, software, validation, data curation, writing - original draft. Ludovica Ceci, methodology, data curation. Rosa Vaccaro, formal analysis. Antonella Di Sotto, data curation, writing - review & editing. Giorgio Vivacqua, formal analysis, writing - review & editing. Rosemarie Heyn, investigation, writing - review & editing. Ludovica Garro, writing - original draft. Marco Tagliafierro, investigation, writing - original draft. Filippo Maria Bassi, methodology. Emanuele Bocci, methodology. Andrea Bassi, formal analysis. Luigi Pannarale, supervision. Antonio Franchitto, writing - review & editing. Paolo Onori, conceptualization, writing - review & editing. Eugenio Gaudio, conceptualization, resources, supervision, funding acquisition.
Supporting Agencies
Sapienza University of RomeData Availability Statement
The datasets used and/or analysed during the current study are available upon reasonable request to the corresponding author.
How to Cite
1.
Mancinelli R, Casini A, Leone S, Ceci L, Vaccaro R, Di Sotto A, et al. Discovering ferroptosis in biliary epithelium during liver fibrosis. Eur J Histochem [Internet]. 2026 Oct. 28 [cited 2026 Oct. 30];70(4). Available from: https://www.ejh.it/ejh/article/view/4615
Copyright (c) 2026 The Author(s)
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
