Schmitt 2022 EBEC

From Bioblast
Jump to navigation Jump to search
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.

Link: EBEC 2022 Conference website

Schmitt Sabine, Gnaiger Erich (2022)

Event: EBEC2022 Marseille FR

Molecular hydrogen H2 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, H2 is typically administered repeatedly or over longer periods of time (hours to days) via inhalation of H2 gas, drinking H2-rich water, or injection of H2 saline, wherefore the observed effects, e.g. on mitochondrial metabolism [2], might be either directly or indirectly related to H2. To investigate a direct short-term effect of H2 on mitochondrial function, we measured mitochondrial respiration and H2O2 production in permeabilized HEK 293T cells upon sequential changes of H2 concentration cH2 in the experimental medium. O2 and H2O2 flux were measured simultaneously in the O2k with the Fluo-Module (Oroboros Instruments). Increase of cH2 was accomplished by injecting H2 into the gas phase of the open O2k-chamber. This causes not only an increase of cH2 but also a decrease of oxygen concentration cO2. As mitochondrial ROS production is a continuous function of cO2, we used the conventionally applied N2 gas as a control to distinguish between cO2- and cH2-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 O2 and H2O2 flux in the LEAK state (without ADP). Upon transition of cO2 from ~160 to ~25 µM, a decrease in O2 and H2O2 flux was observed. This was comparable between regimes with increased cH2 or cN2. Further transitions by re-oxygenation and injection of H2 or N2 yielded the same results. Similarly, cO2-dependent changes in mitochondrial respiration and H2O2 production in the OXPHOS state (kinetically saturating [ADP]) were independent of the increase in cH2 or cN2. These results indicate that short-term exposure to increased cH2 does not affect mitochondrial respiration or H2O2 production.


Bioblast editor: Cecatto C O2k-Network Lab: AT Innsbruck Oroboros, AT Innsbruck Gnaiger E, AT Innsbruck MitoFit


Affiliations

Sabine Schmitt1, Erich Gnaiger1
[email protected]
  1. Oroboros Instruments, Innsbruck, Austria


References

  1. Tian Y, Zhang Y, Wang Y, Chen Y, Fan W, Zhou J, Qiao J, Wei Y (2021) Hydrogen, a Novel Therapeutic Molecule, Regulates Oxidative Stress, Inflammation, and Apoptosis. https://doi.org/10.3389/fphys.2021.789507
  2. Gvozdjáková A, Kucharska J, Kura B, Vancova O, Rausová Z, Sumbalová Z, Uličná O, Slezák J (2020) A new insight into the molecular hydrogen effect on coenzyme Q and mitochondrial function of rats. https://doi.org/10.1139/cjpp-2019-0281

List of abbreviations, terms and definitions - MitoPedia

» MitoPedia: Terms and abbreviations


Labels: MiParea: Respiration 

Stress:Hypoxia  Organism: Human  Tissue;cell: HEK  Preparation: Permeabilized cells 


Coupling state: LEAK, OXPHOS  Pathway:HRR: Oxygraph-2k, O2k-Fluorometer