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| {{Abstract
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| |title=[[File:SchmittS.jpg|left|100px|Sabine Schmitt]]Schmitt S, Gnaiger E (2022) Short-term effect of molecular hydrogen on mitochondrial respiration and hydrogen peroxide production in permeabilized HEK 293T cells. EBEC 2022.
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| |info=[https://ebec2022.org/ EBEC 2022 Conference website]
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| |authors=Schmitt Sabine, Gnaiger Erich
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| |year=2022
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| |event=EBEC2022 Marseille FR
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| |abstract=Molecular hydrogen H<sub>2</sub> has been reported to be an antioxidative, anti-inflammatory, and antiapoptotic agent with therapeutic potential for various diseases such as cardiac arrest, asthma, chronic obstructive pulmonary disease (COPD), and, most recently, COVID-19 [1]. In previous studies, H<sub>2</sub> is typically administered repeatedly or over longer periods of time (hours to days) via inhalation of H<sub>2</sub> gas, drinking H<sub>2</sub>-rich water, or injection of H<sub>2</sub> saline, wherefore the observed effects, e.g. on mitochondrial metabolism [2], might be either directly or indirectly related to H<sub>2</sub>.
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| To investigate a direct short-term effect of H<sub>2</sub> on mitochondrial function, we measured mitochondrial respiration and H<sub>2</sub>O<sub>2</sub> production in permeabilized HEK 293T cells upon sequential changes of H<sub>2</sub> concentration ''c''<sub>H2</sub> in the experimental medium. O<sub>2</sub> and H<sub>2</sub>O<sub>2</sub> flux were measured simultaneously in the O2k with the Fluo-Module (Oroboros Instruments). Increase of ''c''<sub>H2</sub> was accomplished by injecting H<sub>2</sub> into the gas phase of the open O2k-chamber. This causes not only an increase of ''c''<sub>H2</sub> but also a decrease of oxygen concentration ''c''<sub>O2</sub>. As mitochondrial ROS production is a continuous function of ''c''<sub>O2</sub>, we used the conventionally applied N<sub>2</sub> gas as a control to distinguish between ''c''<sub>O2</sub>- and ''c''<sub>H2</sub>-dependent effects. Measurements were started near air saturation (~160 µM of oxygen). The plasma membrane was permeabilized with digitonin and the NADH-linked substrates pyruvate & malate were titrated to measure O<sub>2</sub> and H<sub>2</sub>O<sub>2</sub> flux in the LEAK state (without ADP). Upon transition of ''c''<sub>O2</sub> from ~160 to ~25 µM, a decrease in O<sub>2</sub> and H<sub>2</sub>O<sub>2</sub> flux was observed. This was comparable between regimes with increased ''c''<sub>H2</sub> or ''c''<sub>N2</sub>. Further transitions by re-oxygenation and injection of H<sub>2</sub> or N<sub>2</sub> yielded the same results. Similarly, ''c''<sub>O2</sub>-dependent changes in mitochondrial respiration and H<sub>2</sub>O<sub>2</sub> production in the OXPHOS state (kinetically saturating [ADP]) were independent of the increase in ''c''<sub>H2</sub> or ''c''<sub>N2</sub>. These results indicate that short-term exposure to increased ''c''<sub>H2</sub> does not affect mitochondrial respiration or H<sub>2</sub>O<sub>2</sub> production.
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| |editor=[[Plangger M]]
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| |mipnetlab=AT Innsbruck Oroboros, AT Innsbruck Gnaiger E, AT Innsbruck MitoFit
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| }}
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| {{Labeling
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| |area=Respiration
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| |organism=Human
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| |tissues=HEK
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| |preparations=Permeabilized cells
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| |couplingstates=LEAK, OXPHOS
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| |pathways=N
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| |instruments=Oxygraph-2k, O2k-Fluorometer
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| }}
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| == Affiliations ==
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| == References ==
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| ::::#Y. Tian, Y. Zhang, Y. Wang, Y. Chen, Hydrogen, a Novel Therapeutic Molecule, Regulates Oxidative Stress, Inflammation, and Apoptosis, Frontiers in Physiology, 12 (2021) 1-14
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| ::::#A. Gvozdjáková, J. Kucharska, B. Kura, O. Vancova O, A new insight into the molecular hydrogen effect on coenzyme Q and mitochondrial function of rats, J Physiol Pharmacol., 1 (2020) 29-34
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| == List of abbreviations, terms and definitions - MitoPedia ==
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| {{Template:List of abbreviations, terms and definitions - MitoPedia}}
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