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Role of hepcidin in oxidative stress and cell death of cultured mouse renal collecting duct cells: protection against iron and sensitization to cadmium

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Title: Role of hepcidin in oxidative stress and cell death of cultured mouse renal collecting duct cells: protection against iron and sensitization to cadmium
Authors: Rachel P. L. van Swelm, Stephanie Probst, Eleni Roussa, Natascha A. Wolff, Wing-Kee Lee, Johannes Fels, Bettina Scharner, Frank Thévenod
Source: Arch Toxicol
Archives of Toxicology, 95, 8, pp. 2719-2735
Publisher Information: Springer Science and Business Media LLC, 2021.
Publication Year: 2021
Subject Terms: Male, 0301 basic medicine, Iron, Apoptosis, Deferoxamine, Binding, Competitive, Cell Line, Mice, 03 medical and health sciences, Hepcidins, Molecular Toxicology, Animals, Gene Silencing, Kidney Tubules, Collecting, Cells, Cultured, 0303 health sciences, Binding, Competitive [MeSH], Mice, Inbred C57BL [MeSH], Cell Line [MeSH], Cadmium/toxicity [MeSH], Kidney Tubules, Collecting/drug effects [MeSH], Cell Death/drug effects [MeSH], Deferoxamine/pharmacology [MeSH], Kidney Tubules, Collecting/cytology [MeSH], Male [MeSH], DMT1, DFO, Cadmium/administration, Iron/administration, Female [MeSH], Iron/toxicity [MeSH], Metallothionein, Reactive Oxygen Species/metabolism [MeSH], Hepcidins/genetics [MeSH], Apoptosis/drug effects [MeSH], Animals [MeSH], Gene Silencing [MeSH], Mice [MeSH], Binding Sites [MeSH], Ca, Oxidative Stress/drug effects [MeSH], Cells, Cultured [MeSH], Binding Sites, Cell Death, Radboudumc 11: Renal disorders RIMLS: Radboud Institute for Molecular Life Sciences, Mice, Inbred C57BL, Oxidative Stress, 13. Climate action, Female, Reactive Oxygen Species, Cadmium
Description: The liver hormone hepcidin regulates systemic iron homeostasis. Hepcidin is also expressed by the kidney, but exclusively in distal nephron segments. Several studies suggest hepcidin protects against kidney damage involving Fe2+ overload. The nephrotoxic non-essential metal ion Cd2+ can displace Fe2+ from cellular biomolecules, causing oxidative stress and cell death. The role of hepcidin in Fe2+ and Cd2+ toxicity was assessed in mouse renal cortical [mCCD(cl.1)] and inner medullary [mIMCD3] collecting duct cell lines. Cells were exposed to equipotent Cd2+ (0.5–5 μmol/l) and/or Fe2+ (50–100 μmol/l) for 4–24 h. Hepcidin (Hamp1) was transiently silenced by RNAi or overexpressed by plasmid transfection. Hepcidin or catalase expression were evaluated by RT-PCR, qPCR, immunoblotting or immunofluorescence microscopy, and cell fate by MTT, apoptosis and necrosis assays. Reactive oxygen species (ROS) were detected using CellROX™ Green and catalase activity by fluorometry. Hepcidin upregulation protected against Fe2+-induced mIMCD3 cell death by increasing catalase activity and reducing ROS, but exacerbated Cd2+-induced catalase dysfunction, increasing ROS and cell death. Opposite effects were observed with Hamp1 siRNA. Similar to Hamp1 silencing, increased intracellular Fe2+ prevented Cd2+ damage, ROS formation and catalase disruption whereas chelation of intracellular Fe2+ with desferrioxamine augmented Cd2+ damage, corresponding to hepcidin upregulation. Comparable effects were observed in mCCD(cl.1) cells, indicating equivalent functions of renal hepcidin in different collecting duct segments. In conclusion, hepcidin likely binds Fe2+, but not Cd2+. Because Fe2+ and Cd2+ compete for functional binding sites in proteins, hepcidin affects their free metal ion pools and differentially impacts downstream processes and cell fate.
Document Type: Article
Other literature type
File Description: pdf
Language: English
ISSN: 1432-0738
0340-5761
DOI: 10.1007/s00204-021-03106-z
Access URL: https://link.springer.com/content/pdf/10.1007/s00204-021-03106-z.pdf
https://pubmed.ncbi.nlm.nih.gov/34181029
https://hdl.handle.net/https://repository.ubn.ru.nl/handle/2066/238678
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8298330
https://pubmed.ncbi.nlm.nih.gov/34181029/
https://link.springer.com/article/10.1007/s00204-021-03106-z
https://www.ncbi.nlm.nih.gov/pubmed/34181029
https://europepmc.org/article/MED/34181029
https://link.springer.com/content/pdf/10.1007/s00204-021-03106-z.pdf
https://pub.uni-bielefeld.de/record/2956164
https://doi.org/10.1007/s00204-021-03106-z
https://repository.publisso.de/resource/frl:6451671
Rights: CC BY
"In Copyright" Rights Statement
URL: http://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (http://creativecommons.org/licenses/by/4.0/) .
Accession Number: edsair.doi.dedup.....c8dbdb654c10d20d84c7e04e916550e9
Database: OpenAIRE
Description
Abstract:The liver hormone hepcidin regulates systemic iron homeostasis. Hepcidin is also expressed by the kidney, but exclusively in distal nephron segments. Several studies suggest hepcidin protects against kidney damage involving Fe2+ overload. The nephrotoxic non-essential metal ion Cd2+ can displace Fe2+ from cellular biomolecules, causing oxidative stress and cell death. The role of hepcidin in Fe2+ and Cd2+ toxicity was assessed in mouse renal cortical [mCCD(cl.1)] and inner medullary [mIMCD3] collecting duct cell lines. Cells were exposed to equipotent Cd2+ (0.5–5 μmol/l) and/or Fe2+ (50–100 μmol/l) for 4–24 h. Hepcidin (Hamp1) was transiently silenced by RNAi or overexpressed by plasmid transfection. Hepcidin or catalase expression were evaluated by RT-PCR, qPCR, immunoblotting or immunofluorescence microscopy, and cell fate by MTT, apoptosis and necrosis assays. Reactive oxygen species (ROS) were detected using CellROX™ Green and catalase activity by fluorometry. Hepcidin upregulation protected against Fe2+-induced mIMCD3 cell death by increasing catalase activity and reducing ROS, but exacerbated Cd2+-induced catalase dysfunction, increasing ROS and cell death. Opposite effects were observed with Hamp1 siRNA. Similar to Hamp1 silencing, increased intracellular Fe2+ prevented Cd2+ damage, ROS formation and catalase disruption whereas chelation of intracellular Fe2+ with desferrioxamine augmented Cd2+ damage, corresponding to hepcidin upregulation. Comparable effects were observed in mCCD(cl.1) cells, indicating equivalent functions of renal hepcidin in different collecting duct segments. In conclusion, hepcidin likely binds Fe2+, but not Cd2+. Because Fe2+ and Cd2+ compete for functional binding sites in proteins, hepcidin affects their free metal ion pools and differentially impacts downstream processes and cell fate.
ISSN:14320738
03405761
DOI:10.1007/s00204-021-03106-z