{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,16]],"date-time":"2026-09-16T12:37:59Z","timestamp":1789562279711,"version":"build-2803163510"},"reference-count":45,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2022,1,25]],"date-time":"2022-01-25T00:00:00Z","timestamp":1643068800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100002997","name":"Nierstichting","doi-asserted-by":"publisher","award":["16OKG04"],"award-info":[{"award-number":["16OKG04"]}],"id":[{"id":"10.13039\/501100002997","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Hemolysis is known to cause acute kidney injury (AKI). The iron regulatory hormone hepcidin, produced by renal distal tubules, is suggested to exert a renoprotective role during this pathology. We aimed to elucidate the molecular mechanisms of renal hepcidin synthesis and its protection against hemoglobin-induced AKI. In contrast to known hepatic hepcidin induction, incubation of mouse cortical collecting duct (mCCDcl1) cells with IL-6 or LPS did not induce Hamp1 mRNA expression, whereas iron (FeS) and hemin significantly induced hepcidin synthesis (p &lt; 0.05). Moreover, iron\/heme-mediated hepcidin induction in mCCDcl1 cells was caused by the nuclear factor erythroid 2-related factor 2 (Nrf2) pathway, as indicated by increased nuclear Nrf2 translocation and induced expression of Nrf2 downstream targets GCLM (p &lt; 0.001), NQO1 (p &lt; 0.001), and TXNRD1 (p &lt; 0.005), which could be prevented by the known Nrf2 inhibitor trigonelline. Newly created inducible kidney-specific hepcidin KO mice demonstrated a significant reduction in renal Hamp1 mRNA expression. Phenylhydrazine (PHZ)-induced hemolysis caused renal iron loading and oxidative stress in both wildtype (Wt) and KO mice. PHZ treatment in Wt induced inflammatory markers (IL-6, TNF\u03b1) but not Hamp1. However, since PHZ treatment also significantly reduced systemic hepcidin levels in both Wt and KO mice (both p &lt; 0.001), a dissection between the roles of systemic and renal hepcidin could not be made. Combined, the results of our study indicate that there are kidney-specific mechanisms in hepcidin regulation, as indicated by the dominant role of iron and not inflammation as an inducer of renal hepcidin, but also emphasize the complex interplay of various iron regulatory mechanisms during AKI on a local and systemic level.<\/jats:p>","DOI":"10.3390\/ijms23031352","type":"journal-article","created":{"date-parts":[[2022,1,25]],"date-time":"2022-01-25T20:40:18Z","timestamp":1643143218000},"page":"1352","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Investigating the Molecular Mechanisms of Renal Hepcidin Induction and Protection upon Hemoglobin-Induced Acute Kidney Injury"],"prefix":"10.3390","volume":"23","author":[{"given":"Laura E.","family":"Diepeveen","sequence":"first","affiliation":[{"name":"Translational Metabolic Laboratory (TML-830), Department of Laboratory Medicine, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, 6525 GA Nijmegen, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gaby","family":"Stegemann","sequence":"additional","affiliation":[{"name":"Translational Metabolic Laboratory (TML-830), Department of Laboratory Medicine, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, 6525 GA Nijmegen, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Erwin T.","family":"Wiegerinck","sequence":"additional","affiliation":[{"name":"Translational Metabolic Laboratory (TML-830), Department of Laboratory Medicine, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, 6525 GA Nijmegen, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rian","family":"Roelofs","sequence":"additional","affiliation":[{"name":"Translational Metabolic Laboratory (TML-830), Department of Laboratory Medicine, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, 6525 GA Nijmegen, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Myrthe","family":"Naber","sequence":"additional","affiliation":[{"name":"Translational Metabolic Laboratory (TML-830), Department of Laboratory Medicine, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, 6525 GA Nijmegen, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Olivier","family":"L\u00f3real","sequence":"additional","affiliation":[{"name":"INSERM 1241, Nutrition, Metabolisms and Cancer (NuMeCan) Institute, University of Rennes, 35033 Rennes, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bart","family":"Smeets","sequence":"additional","affiliation":[{"name":"Department of Pathology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, 6525 GA Nijmegen, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8663-3498","authenticated-orcid":false,"given":"Frank","family":"Th\u00e9venod","sequence":"additional","affiliation":[{"name":"Institute of Physiology, Pathophysiology and Toxicology, Center for Biomedical Training and Research (ZBAF), University of Witten\/Herdecke, 58455 Witten, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1040-9446","authenticated-orcid":false,"given":"Dorine W.","family":"Swinkels","sequence":"additional","affiliation":[{"name":"Translational Metabolic Laboratory (TML-830), Department of Laboratory Medicine, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, 6525 GA Nijmegen, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rachel P. L.","family":"van Swelm","sequence":"additional","affiliation":[{"name":"Translational Metabolic Laboratory (TML-830), Department of Laboratory Medicine, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, 6525 GA Nijmegen, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,1,25]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"813","DOI":"10.1001\/jama.294.7.813","article-title":"Acute renal failure in critically ill patients: A multinational, multicenter study","volume":"294","author":"Uchino","year":"2005","journal-title":"JAMA"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"34","DOI":"10.1159\/000361062","article-title":"Cardiac surgery-associated acute kidney injury","volume":"37","author":"Mao","year":"2014","journal-title":"Blood Purif."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"257","DOI":"10.1051\/ject\/200840257","article-title":"Hemolysis in cardiac surgery patients undergoing cardiopulmonary bypass: A review in search of a treatment algorithm","volume":"40","author":"Vercaemst","year":"2008","journal-title":"J. Extra-Corpor. Technol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"103","DOI":"10.1016\/0014-5793(93)80974-Y","article-title":"Chelatable iron and copper can be released from extracorporeally circulated blood during cardiopulmonary bypass","volume":"328","author":"Moat","year":"1993","journal-title":"FEBS Lett."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1276","DOI":"10.1182\/blood-2012-11-451229","article-title":"Hemolysis and free hemoglobin revisited: Exploring hemoglobin and hemin scavengers as a novel class of therapeutic proteins","volume":"121","author":"Schaer","year":"2013","journal-title":"Blood"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"2024","DOI":"10.1016\/j.jacc.2009.12.046","article-title":"Novel biomarkers, oxidative stress, and the role of labile iron toxicity in cardiopulmonary bypass-associated acute kidney injury","volume":"55","author":"Haase","year":"2010","journal-title":"J. Am. Coll. Cardiol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"423","DOI":"10.1681\/ASN.V132423","article-title":"Megalin and cubilin are endocytic receptors involved in renal clearance of hemoglobin","volume":"13","author":"Gburek","year":"2002","journal-title":"J. Am. Soc. Nephrol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"550","DOI":"10.1038\/s41419-018-0568-z","article-title":"Endogenous hepcidin synthesis protects the distal nephron against hemin and hemoglobin mediated necroptosis","volume":"9","author":"Vos","year":"2018","journal-title":"Cell Death Dis."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1097\/01.mnh.0000437613.88158.d3","article-title":"Heme oxygenase-1 and acute kidney injury","volume":"23","author":"Nath","year":"2014","journal-title":"Curr. Opin. Nephrol. Hypertens."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"3091","DOI":"10.1111\/j.1582-4934.2008.00494.x","article-title":"Hepcidin suppression and defective iron recycling account for dysregulation of iron homeostasis in heme oxygenase-1 deficiency","volume":"13","author":"Kartikasari","year":"2009","journal-title":"J. Cell. Mol. Med."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1480","DOI":"10.1016\/j.freeradbiomed.2011.02.011","article-title":"Postoperative acute kidney injury is associated with hemoglobinemia and an enhanced oxidative stress response","volume":"50","author":"Billings","year":"2011","journal-title":"Free Radic. Biol. Med."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"2340","DOI":"10.2215\/CJN.01000211","article-title":"Urinary hepcidin-25 and risk of acute kidney injury following cardiopulmonary bypass","volume":"6","author":"Ho","year":"2011","journal-title":"Clin. J. Am. Soc. Nephrol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"595","DOI":"10.1093\/ndt\/gfr387","article-title":"Greater increase in urinary hepcidin predicts protection from acute kidney injury after cardiopulmonary bypass","volume":"27","author":"Prowle","year":"2012","journal-title":"Nephrol. Dial. Transplant."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"R186","DOI":"10.1186\/cc10339","article-title":"Urine hepcidin has additive value in ruling out cardiopulmonary bypass-associated acute kidney injury: An observational cohort study","volume":"15","author":"Mertens","year":"2011","journal-title":"Crit. Care"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"2720","DOI":"10.1681\/ASN.2015040461","article-title":"Renal Handling of Circulating and Renal-Synthesized Hepcidin and Its Protective Effects against Hemoglobin-Mediated Kidney Injury","volume":"27","author":"Wetzels","year":"2016","journal-title":"J. Am. Soc. Nephrol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"361","DOI":"10.1677\/joe.1.05729","article-title":"The iron-regulatory peptide hormone hepcidin: Expression and cellular localization in the mammalian kidney","volume":"184","author":"Kulaksiz","year":"2005","journal-title":"J. Endocrinol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"2800","DOI":"10.1681\/ASN.2014101037","article-title":"Hepcidin Mitigates Renal Ischemia-Reperfusion Injury by Modulating Systemic Iron Homeostasis","volume":"26","author":"Scindia","year":"2015","journal-title":"J. Am. Soc. Nephrol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1864","DOI":"10.1182\/blood-2005-03-1159","article-title":"Time-course analysis of hepcidin, serum iron, and plasma cytokine levels in humans injected with LPS","volume":"106","author":"Kemna","year":"2005","journal-title":"Blood"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"519","DOI":"10.1038\/s42255-019-0063-6","article-title":"Nrf2 controls iron homeostasis in haemochromatosis and thalassaemia via Bmp6 and hepcidin","volume":"1","author":"Lim","year":"2019","journal-title":"Nat. Metab."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"93","DOI":"10.1016\/j.freeradbiomed.2015.06.006","article-title":"Molecular basis of the Keap1-Nrf2 system","volume":"88","author":"Suzuki","year":"2015","journal-title":"Free Radic. Biol. Med."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1192","DOI":"10.1016\/j.freeradbiomed.2015.11.001","article-title":"Phytoestrogens modulate hepcidin expression by Nrf2: Implications for dietary control of iron absorption","volume":"89","author":"Bayele","year":"2015","journal-title":"Free Radic. Biol. Med."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"105701","DOI":"10.1016\/j.envint.2020.105701","article-title":"Polychlorinated biphenyl quinone induces hepatocytes iron overload through up-regulating hepcidin expression","volume":"139","author":"Liu","year":"2020","journal-title":"Environ. Int."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"179","DOI":"10.1016\/j.toxlet.2018.06.1218","article-title":"Inhibition of Nrf2 alters cell stress induced by chronic iron exposure in human proximal tubular epithelial cells","volume":"295","author":"Masereeuw","year":"2018","journal-title":"Toxicol. Lett."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"323","DOI":"10.1146\/annurev.pharmtox.010909.105600","article-title":"Mechanisms of cell protection by heme oxygenase-1","volume":"50","author":"Gozzelino","year":"2010","journal-title":"Annu. Rev. Pharm. Toxicol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"197","DOI":"10.1146\/annurev.nutr.28.061807.155521","article-title":"Systemic Iron Homeostasis and the Iron-Responsive Element\/Iron-Regulatory Protein (IRE\/IRP) Regulatory Network","volume":"28","author":"Muckenthaler","year":"2008","journal-title":"Annu. Rev. Nutr."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"473","DOI":"10.1159\/000475890","article-title":"Targeting the KEAP1-NRF@ system to prevent kidney disease progression","volume":"45","author":"Nezu","year":"2017","journal-title":"Am. J. Nephrol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"3055","DOI":"10.2337\/db11-0807","article-title":"Therapeutic potential of Nrf2 activators in streptozotocin-induced diabetic nephropathy","volume":"60","author":"Zheng","year":"2011","journal-title":"Diabetes"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1960","DOI":"10.2353\/ajpath.2006.051113","article-title":"Multiorgan autoimmune inflammation, enhanced lymphoproliferation, and impaired homeostasis of reactive oxygen species in mice lacking the antioxidant-activated transcription factor Nrf2","volume":"168","author":"Ma","year":"2006","journal-title":"Am. J. Pathol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"387","DOI":"10.1016\/j.kint.2016.08.023","article-title":"Transcription factor Nrf2 hyperactivation in early-phase renal ischemia-reperfusion injury prevents tubular damage progression","volume":"91","author":"Nezu","year":"2017","journal-title":"Kidney Int."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"327","DOI":"10.1056\/NEJMoa1105351","article-title":"Bardoxolone methyl and kidney function in CKD with type 2 diabetes","volume":"365","author":"Pergola","year":"2011","journal-title":"N. Engl. J. Med."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1756","DOI":"10.1089\/ars.2017.7176","article-title":"The Roles of NRF2 in Modulating Cellular Iron Homeostasis","volume":"29","author":"Kerins","year":"2018","journal-title":"Antioxid. Redox Signal."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"433","DOI":"10.1089\/ars.2020.8148","article-title":"NRF2 and Hypoxia-Inducible Factors: Key Players in the Redox Control of Systemic Iron Homeostasis","volume":"35","author":"Duarte","year":"2020","journal-title":"Antioxid. Redox Signal."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"299","DOI":"10.1016\/j.mam.2005.07.001","article-title":"Iron, oxidative stress and human health","volume":"26","author":"Puntarulo","year":"2005","journal-title":"Mol. Asp. Med."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1069","DOI":"10.1016\/j.ajpath.2010.12.005","article-title":"Loss of endogenous bone morphogenetic protein-6 aggravates renal fibrosis","volume":"178","author":"Dendooven","year":"2011","journal-title":"Am. J. Pathol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"F1317","DOI":"10.1152\/ajprenal.00030.2015","article-title":"Tubular cross talk in acute kidney injury: A story of sense and sensibility","volume":"308","author":"Dagher","year":"2015","journal-title":"Am. J. Physiol. Ren. Physiol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1271","DOI":"10.1172\/JCI200420945","article-title":"IL-6 mediates hypoferremia of inflammation by inducing the synthesis of the iron regulatory hormone hepcidin","volume":"113","author":"Nemeth","year":"2004","journal-title":"J. Clin. Investig."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"979","DOI":"10.1038\/nm.1865","article-title":"An efficient and versatile system for acute and chronic modulation of renal tubular function in transgenic mice","volume":"14","author":"Greiner","year":"2008","journal-title":"Nat. Med."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"e35","DOI":"10.1097\/HS9.0000000000000035","article-title":"Erythroferrone: An Erythroid Regulator of Hepcidin and Iron Metabolism","volume":"2","author":"Coffey","year":"2018","journal-title":"HemaSphere"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"3039","DOI":"10.1182\/blood-2010-02-270322","article-title":"Hepatic HIF-2 regulates erythropoietic responses to hypoxia in renal anemia","volume":"116","author":"Kapitsinou","year":"2010","journal-title":"Blood"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1143","DOI":"10.3324\/haematol.2018.208645","article-title":"The opposing effects of acute inflammation and iron deficiency anemia on serum hepcidin and iron absorption in young women","volume":"104","author":"Stoffel","year":"2019","journal-title":"Haematologica"},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Yousef Yengej, F.A., Jansen, J., Rookmaaker, M.B., Verhaar, M.C., and Clevers, H. (2020). Kidney Organoids and Tubuloids. Cells, 9.","DOI":"10.3390\/cells9061326"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"878","DOI":"10.1681\/ASN.2004121110","article-title":"Mineralocorticoid versus glucocorticoid receptor occupancy mediating aldosterone-stimulated sodium transport in a novel renal cell line","volume":"16","author":"Gaeggeler","year":"2005","journal-title":"J. Am. Soc. Nephrol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1966","DOI":"10.1681\/ASN.2013010018","article-title":"Albumin is recycled from the primary urine by tubular transcytosis","volume":"24","author":"Tenten","year":"2013","journal-title":"J. Am. Soc. Nephrol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"3646","DOI":"10.1182\/blood-2014-01-550467","article-title":"Targeted disruption of hepcidin in the liver recapitulates the hemochromatotic phenotype","volume":"123","author":"Zumerle","year":"2014","journal-title":"Blood"},{"key":"ref_45","first-page":"9","article-title":"A simple technique for measuring storage iron concentrations in formalinized liver samples","volume":"33","author":"Torrance","year":"1968","journal-title":"S. Afr. J. Med. Sci."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/3\/1352\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T22:07:28Z","timestamp":1760134048000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/3\/1352"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,1,25]]},"references-count":45,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2022,2]]}},"alternative-id":["ijms23031352"],"URL":"https:\/\/doi.org\/10.3390\/ijms23031352","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,1,25]]}}}