Agrimoniin Alleviates Ferroptosis in Cold-Stored DCD Liver Grafts Through Activation of the Nrf-2 Pathway

Abstract

Liver grafts from donation-after-cardiac-death (DCD) are vulnerable to ischemia–reperfusion injury, which compromises graft function after transplantation. Agrimoniin has been shown to possess antioxidant and anti-inflammatory properties, making it a potential therapeutic agent for organ preservation. This study investigated whether supplementing agrimoniin to the University of Wisconsin (UW) cold storage solution protected liver grafts from DCD rats or cold preserved human liver cell lines (QSG-7701 and HepG2). Agrimoniin supplementation significantly reduced oxidative damage, alleviated ferroptosis, and mitigated liver injury by activating the Nrf-2 pathway, both in vivo and in vitro. These findings suggest that ferroptosis is a mediator in DCD liver injury, and agrimoniin, through its activation of the Nrf-2 pathway, may be an effective therapeutic agent for enhancing liver graft preservation and improving outcomes in DCD liver transplantation.

Keywords

agrimoniin / ferroptosis / liver transplantation / Nrf-2 pathway

References

[1] J. H. Kwon, W. M. Blanding, K. Shorbaji, et al., “Waitlist and Transplant Outcomes in Organ Donation After Circulatory Death: Trends in the United States,” Annals of Surgery 278, no. 4 (2023): 609–620.

[2] N. A. Terrault, C. Francoz, M. Berenguer, M. Charlton, and J. Heimbach, “Liver Transplantation 2023: Status Report, Current and Future Challenges,” Clinical Gastroenterology and Hepatology 21, no. 8 (2023): 2150–2166.

[3] K. Pandya, V. Sastry, M. T. Panlilio, et al., “Differential Impact of Extended Criteria Donors After Brain Death or Circulatory Death in Adult Liver Transplantation,” Liver Transplantation 26, no. 12 (2020): 1603–1617.

[4] T. Zhang, J. Dunson, F. Kanwal, et al., “Trends in Outcomes for Marginal Allografts in Liver Transplant,” JAMA Surgery 155, no. 10 (2020): 926–932.

[5] P. E. Morrissey and A. P. Monaco, “Donation After Circulatory Death: Current Practices, Ongoing Challenges, and Potential Improvements,” Transplantation 97, no. 3 (2014): 258–264.

[6] R. P. H. Meier, Y. Kelly, H. Braun, et al., “Comparison of Biliary Complications Rates After Brain Death, Donation After Circulatory Death, and Living-Donor Liver Transplantation: A Single-Center Cohort Study,” Transplant International 35 (2022): 10855.

[7] R. G. Bardallo, R. T. Da Silva, T. Carbonell, et al., “Liver Graft Hypothermic Static and Oxygenated Perfusion (HOPE) Strategies: A Mitochondrial Crossroads,” International Journal of Molecular Sciences 23, no. 10 (2022): 5742.

[8] M. A. Zaouali, F. Bardag-Gorce, T. Carbonell, et al., “Proteasome Inhibitors Protect the Steatotic and Non-Steatotic Liver Graft Against Cold Ischemia Reperfusion Injury,” Experimental and Molecular Pathology 94, no. 2 (2013): 352–359.

[9] J. P. Faure, I. Petit, K. Zhang, et al., “Protective Roles of Polyethylene Glycol and Trimetazidine Against Cold Ischemia and Reperfusion Injuries of Pig Kidney Graft,” American Journal of Transplantation 4, no. 4 (2004): 495–504.

[10] E. Karlinska, B. Romanowska, and M. Kosmala, “The Aerial Parts of Agrimonia Procera Wallr. And Agrimonia eupatoria L. as a Source of Polyphenols, and Especially Agrimoniin and Flavonoids,” Molecules 26, no. 24 (2021): 7706.

[11] T. N. Santos, G. Costa, J. P. Ferreira, et al., “Antioxidant, Anti-Inflammatory, and Analgesic Activities of Agrimonia eupatoriaL. Infusion,” Evidence-Based Complementary and Alternative Medicine 2017 (2017): 8309894.

[12] K. P. Storves, M. R. Talcott, J. M. Wallace, et al., “The Veterinary Consortium for Research Animal Care and Welfare Survey on Revisions to the Eighth Edition of the Guide for the Care and Use of Laboratory Animals,” Journal of the American Association for Laboratory Animal Science 64, no. 4 (2025): 1–10.

[13] Q. Chen, X. Liu, Z. Liu, et al., “Tackling Regulated Cell Death Yields Enhanced Protection in Lung Grafts,” Theranostics 13, no. 13 (2023): 4376–4390.

[14] Y. Zhai, H. Petrowsky, J. C. Hong, R. W. Busuttil, and J. W. Kupiec-Weglinski, “Ischaemia-Reperfusion Injury in Liver Transplantation—From Bench to Bedside,” Nature Reviews. Gastroenterology & Hepatology 10, no. 2 (2013): 79–89.

[15] G. Chullo, A. Panisello-Rosello, N. Marquez, et al., “Focusing on Ischemic Reperfusion Injury in the New Era of Dynamic Machine Perfusion in Liver Transplantation,” International Journal of Molecular Sciences 25, no. 2 (2024): 1117.

[16] N. Yamada, T. Karasawa, T. Wakiya, et al., “Iron Overload as a Risk Factor for Hepatic Ischemia-Reperfusion Injury in Liver Transplantation: Potential Role of Ferroptosis,” American Journal of Transplantation 20, no. 6 (2020): 1606–1618.

[17] Z. Wu, Y. Zhu, W. Liu, et al., “Ferroptosis in Liver Disease: Natural Active Compounds and Therapeutic Implications,” Antioxidants (Basel) 13, no. 3 (2024): 352.

[18] J. Zhou, J. Chen, Q. Wei, K. Saeb-Parsy, and X. Xu, “The Role of Ischemia/Reperfusion Injury in Early Hepatic Allograft Dysfunction,” Liver Transplantation 26, no. 8 (2020): 1034–1048.

[19] T. Ito, B. V. Naini, D. Markovic, et al., “Ischemia-Reperfusion Injury and Its Relationship With Early Allograft Dysfunction in Liver Transplant Patients,” American Journal of Transplantation 21, no. 2 (2021): 614–625.

[20] S. J. Dixon, K. M. Lemberg, M. R. Lamprecht, et al., “Ferroptosis: An Iron-Dependent Form of Nonapoptotic Cell Death,” Cell 149, no. 5 (2012): 1060–1072.

[21] W. S. Yang and B. R. Stockwell, “Ferroptosis: Death by Lipid Peroxidation,” Trends in Cell Biology 26, no. 3 (2016): 165–176.

[22] K. Bersuker, J. M. Hendricks, Z. Li, et al., “The CoQ Oxidoreductase FSP1 Acts Parallel to GPX4 to Inhibit Ferroptosis,” Nature 575, no. 7784 (2019): 688–692.

[23] V. A. N. Kraft, C. T. Bezjian, S. Pfeiffer, et al., “GTP Cyclohydrolase 1/Tetrahydrobiopterin Counteract Ferroptosis Through Lipid Remodeling,” ACS Central Science 6, no. 1 (2020): 41–53.

[24] F. He, X. Ru, and T. Wen, “NRF2, a Transcription Factor for Stress Response and Beyond,” International Journal of Molecular Sciences 21, no. 13 (2020): 4777.

[25] J. Hoffmann, F. Casetti, U. Bullerkotte, et al., “Anti-Inflammatory Effects of Agrimoniin-Enriched Fractions of Potentilla erecta,” Molecules 21, no. 6 (2016): 792.

[26] T. A. Fedotcheva, O. P. Sheichenko, and N. I. Fedotcheva, “New Properties and Mitochondrial Targets of Polyphenol Agrimoniin as a Natural Anticancer and Preventive Agent,” Pharmaceutics 13, no. 12 (2021): 2089.

[27] T. Jin, L. Chi, and C. Ma, “Agrimonia Pilosa: A Phytochemical and Pharmacological Review,” Evidence-Based Complementary and Alternative Medicine 2022 (2022): 3742208.

[28] A. Bazylko, J. P. Piwowarski, A. Filipek, J. Bonarewicz, and M. Tomczyk, “In Vitro Antioxidant and Anti-Inflammatory Activities of Extracts From Potentilla Recta and Its Main Ellagitannin, Agrimoniin,” Journal of Ethnopharmacology 149, no. 1 (2013): 222–227.

[29] S. B. Kim, S. H. Hwang, H. W. Suh, and S. S. Lim, “Phytochemical Analysis of Agrimonia Pilosa Ledeb, Its Antioxidant Activity and Aldose Reductase Inhibitory Potential,” International Journal of Molecular Sciences 18, no. 2 (2017): 379.

[30] M. Fumagalli, E. Sangiovanni, U. Vrhovsek, et al., “Strawberry Tannins Inhibit IL-8 Secretion in a Cell Model of Gastric Inflammation,” Pharmacological Research 111 (2016): 703–712.

[31] Q. Ma, “Role of nrf2 in Oxidative Stress and Toxicity,” Annual Review of Pharmacology and Toxicology 53 (2013): 401–426.

[32] R. Li, Z. Jia, and H. Zhu, “Regulation of Nrf2 Signaling,” Reactive Oxygen Species (Apex) 8, no. 24 (2019): 312–322.

[33] S. Yuan, C. Wei, G. Liu, et al., “Sorafenib Attenuates Liver Fibrosis by Triggering Hepatic Stellate Cell Ferroptosis via HIF-1α/SLC7A11 Pathway,” Cell Proliferation 55, no. 1 (2022): e13158.

[34] X. Cheng, X. Ge, C. Zhang, et al., “Tryptophan Suppresses FTH1-Driven Ferritinophagy, a Key Correlate of Prognosis in Hepatocellular Carcinoma,” Cell Proliferation 59 (2025): e70074.

[35] D. Ross and D. Siegel, “The Diverse Functionality of NQO1 and Its Roles in Redox Control,” Redox Biology 41 (2021): 101950.

[36] N. F. Villeneuve, A. Lau, and D. D. Zhang, “Regulation of the Nrf2-Keap1 Antioxidant Response by the Ubiquitin Proteasome System: An Insight Into Cullin-Ring Ubiquitin Ligases,” Antioxidants & Redox Signaling 13, no. 11 (2010): 1699–1712.

[37] V. Ngo and M. L. Duennwald, “Nrf2 and Oxidative Stress: A General Overview of Mechanisms and Implications in Human Disease,” Antioxidants (Basel) 11, no. 12 (2022): 2345.

[38] M. R. Voigt and G. T. DeLario, “Perspectives on Abdominal Organ Preservation Solutions: A Comparative Literature Review,” Progress in Transplantation 23, no. 4 (2013): 383–391.

[39] C. Bai, J. Hua, D. Meng, et al., “Glutaminase-1 Mediated Glutaminolysis to Glutathione Synthesis Maintains Redox Homeostasis and Modulates Ferroptosis Sensitivity in Cancer Cells,” Cell Proliferation 58, no. 11 (2025): e70036.

[40] S. Loganathan, T. Radovits, K. Hirschberg, et al., “Effects of Custodiol-N, a Novel Organ Preservation Solution, on Ischemia/Reperfusion Injury,” Journal of Thoracic and Cardiovascular Surgery 139, no. 4 (2010): 1048–1056.

[41] F. Giampieri, J. M. Alvarez-Suarez, L. Mazzoni, et al., “An Anthocyanin-Rich Strawberry Extract Protects Against Oxidative Stress Damage and Improves Mitochondrial Functionality in Human Dermal Fibroblasts Exposed to an Oxidizing Agent,” Food & Function 5, no. 8 (2014): 1939–1948.

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