Figure 10. Pretreatment with Molecular Hydrogen Enhances Mitochondrial Aerobic Metabolism.
ImBPCs were subjected to a mitochondrial stress test after treatment with molecular hydrogen water (red) or control (dI water; blue) for 24 h. Oxygen consumption rates (OCR) (Panel A) and extracellular acidification rates (ECAR) (Panel C) were measured using the Seahorse XF24 analyzer in the presence of the ATP synthase inhibitor (oligomycin; 3 µM), the uncoupling agent (FCCP; 3 µM), the complex I inhibitor (rotenone; 3 µM), and the complex III inhibitor (antimycin A; 1.5 µM). Analysis of OCR data showed molecular hydrogen increased basal respiration, reserve capacity, and non-mitochondrial (non-mt) respiration but had no effects on ATP production rate as assessed indirectly by OCR analysis, proton leak, or maximal respiration (Panel B). Direct measurement of ATP levels (Panel D) in other cells not exposed to the metabolic stressors showed higher levels of ATP after 24 h but not after 10 min or 6 h of exposure to molecular hydrogen. Results presented here are from one experiment completed, which is representative of the 3 experiments that were done. Results are represented as a mean ± SEM. ** is p<0.01 and *** is p<0.001.