Mitochondrial electron transport chain is essential for acute inflammatory stress responses in mouse fibroblasts

Marie M. Mühlon; Christina Schenkl; Lukas Harder; Luca Giordano; Julian M. Voll; Christian Franke; Diana Dudziak; Torsten Doenst; Ralf A. Claus; Marten Szibor

doi:10.1038/s41598-026-61303-1

Mitochondrial electron transport chain is essential for acute inflammatory stress responses in mouse fibroblasts

Marie M. Mühlon
, Christina Schenkl
, Lukas Harder
, Luca Giordano
, Julian M. Voll
, Christian Franke
, Diana Dudziak
, Torsten Doenst
, Ralf A. Claus
, Marten Szibor^*

^*Corresponding author for this work

Biosciences

Research output: Contribution to journal › Article › Scientific › peer-review

Abstract

Fibroblasts are linked to stress responses in a broad number of diseases. Here, we used immortalized mouse embryonic fibroblasts (iMEFs) to elucidate their signaling behavior in response to proinflammatory lipopolysaccharides (LPS) and angiotensin II (Ang-II). To test for the role of the mitochondrial electron transport chain (ETC), iMEFs were cultured in glucose- and galactose-containing media promoting glycolysis and mitochondrial oxidative phosphorylation, respectively. In addition, we used alternative oxidase (AOX), a ubiquinol oxidoreductase that serves as a naturally evolved rescue mechanism in case of ETC disruption. We found that within 24 h of treatment, LPS upregulated a number of proinflammatory genes, namely Tlr4, Il6, Tgfb1, Nlrp3, Casp1, and Il1b; largely, the effect was more pronounced in galactose-containing media and attenuated by AOX. The increase in transcripts resulted partly in elevated cytokine secretion. Twenty-four hours of Ang-II treatment also induced these genes, albeit to a lesser degree and less sensitive to AOX. Cellular oxygen consumption rates (OCRs) were higher in galactose media but remained unaffected by either stimulus. Our results suggest that fibroblasts undergo a similar proinflammatory phenotypic shift in response to different stressors. This response is shaped by ETC activity, which, surprisingly, is not reflected in altered OCRs.

Original language	English
Article number	24156
Journal	Scientific Reports
Volume	16
Issue number	1
DOIs	https://doi.org/10.1038/s41598-026-61303-1
Publication status	Published - 2026
Publication type	A1 Journal article-refereed

Funding

Open Access funding enabled and organized by Projekt DEAL. MSZ was supported by intramural funds (LOM) from the Jena University Hospital. MM received a scholarship from the Interdisciplinary Center for Clinical Research (IZKF). DD is principal investigator Germany’s Excellence Strategy – EXC 2051 Balance of the Microverse – Project-ID 390713860. CF is supported by the Thüringer Aufbaubank (TAB, FGR 0060 KI-supER) and the DFG via the Collaborative Research Center PolyTarget (CRC 1278, project number 316213987, project B07). The authors thank the Microverse Imaging Center for providing microscope facility support. The Microverse Imaging Center is funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) under Germany’s Excellence Strategy – EXC 2051 – Project-ID 39071386 and project number 316213987 (CRC 1278, project Z01). The ELYRA 7 was funded by the Free State of Thuringia with grant number 2019 FGI 0003. The authors thank Judith Himmel (Dept. of Cardiothoracic Surgery, Jena University Hospital) and Katrin Hornung (Core Facility Cytometry, Institute of Immunology, Jena University Hospital) for excellent technical support. Furthermore, the authors would like to express their deep gratitude to Howard T. Jacobs (Tampere University) for providing critical materials and always insightful discussions. All authors are indebted to Ryan Titmas of Lucid Scientific (Boston, MA, USA) for providing the Resipher. MSZ was supported by intramural funds (LOM). MM received a scholarship from the Interdisciplinary Center for Clinical Research (IZKF). DD is principal investigator Germany’s Excellence Strategy – EXC 2051 Balance of the Microverse – Project-ID 390713860. C.F. is supported by the Thüringer Aufbaubank (TAB, FGR 0060 KI-supER) and the DFG via the Collaborative Research Center PolyTarget (CRC 1278, project number 316213987, project B07). We thank the Microverse Imaging Center for providing microscope facility support. The Microverse Imaging Center is funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) under Germany’s Excellence Strategy – EXC 2051 – Project-ID 39071386 and project number 316213987 (CRC 1278, project Z01). The ELYRA 7 was funded by the Free State of Thuringia with grant number 2019 FGI 0003.

Keywords

Alternative oxidase
Electron transport chain
Fibroblasts
Inflammatory response
Mitochondria
Mouse

Publication forum classification

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ASJC Scopus subject areas

General

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https://urn.fi/URN:NBN:fi:tuni-202608249227Licence: CC BY

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title = "Mitochondrial electron transport chain is essential for acute inflammatory stress responses in mouse fibroblasts",

abstract = "Fibroblasts are linked to stress responses in a broad number of diseases. Here, we used immortalized mouse embryonic fibroblasts (iMEFs) to elucidate their signaling behavior in response to proinflammatory lipopolysaccharides (LPS) and angiotensin II (Ang-II). To test for the role of the mitochondrial electron transport chain (ETC), iMEFs were cultured in glucose- and galactose-containing media promoting glycolysis and mitochondrial oxidative phosphorylation, respectively. In addition, we used alternative oxidase (AOX), a ubiquinol oxidoreductase that serves as a naturally evolved rescue mechanism in case of ETC disruption. We found that within 24 h of treatment, LPS upregulated a number of proinflammatory genes, namely Tlr4, Il6, Tgfb1, Nlrp3, Casp1, and Il1b; largely, the effect was more pronounced in galactose-containing media and attenuated by AOX. The increase in transcripts resulted partly in elevated cytokine secretion. Twenty-four hours of Ang-II treatment also induced these genes, albeit to a lesser degree and less sensitive to AOX. Cellular oxygen consumption rates (OCRs) were higher in galactose media but remained unaffected by either stimulus. Our results suggest that fibroblasts undergo a similar proinflammatory phenotypic shift in response to different stressors. This response is shaped by ETC activity, which, surprisingly, is not reflected in altered OCRs.",

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AU - Giordano, Luca

AU - Voll, Julian M.

AU - Franke, Christian

AU - Dudziak, Diana

AU - Doenst, Torsten

AU - Claus, Ralf A.

AU - Szibor, Marten

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AB - Fibroblasts are linked to stress responses in a broad number of diseases. Here, we used immortalized mouse embryonic fibroblasts (iMEFs) to elucidate their signaling behavior in response to proinflammatory lipopolysaccharides (LPS) and angiotensin II (Ang-II). To test for the role of the mitochondrial electron transport chain (ETC), iMEFs were cultured in glucose- and galactose-containing media promoting glycolysis and mitochondrial oxidative phosphorylation, respectively. In addition, we used alternative oxidase (AOX), a ubiquinol oxidoreductase that serves as a naturally evolved rescue mechanism in case of ETC disruption. We found that within 24 h of treatment, LPS upregulated a number of proinflammatory genes, namely Tlr4, Il6, Tgfb1, Nlrp3, Casp1, and Il1b; largely, the effect was more pronounced in galactose-containing media and attenuated by AOX. The increase in transcripts resulted partly in elevated cytokine secretion. Twenty-four hours of Ang-II treatment also induced these genes, albeit to a lesser degree and less sensitive to AOX. Cellular oxygen consumption rates (OCRs) were higher in galactose media but remained unaffected by either stimulus. Our results suggest that fibroblasts undergo a similar proinflammatory phenotypic shift in response to different stressors. This response is shaped by ETC activity, which, surprisingly, is not reflected in altered OCRs.

KW - Alternative oxidase

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KW - Mitochondria

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Mitochondrial electron transport chain is essential for acute inflammatory stress responses in mouse fibroblasts

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