Sumbalova 2017 MiP2017
Human blood cells are an attractive source of mitochondria for research and diagnostic purposes. In our study we evaluated the effect of regular physical activity on respiration of isolated peripheral blood mononuclear cells (PBMC) and platelets (PLT).
The active group of 15 healthy participants (10 men, 5 women) with age 39-64 years participated in regular supervised training once a week for several years. This group was age-matched with the inactive group of 15 participants without any regular physical activity for several years (9 men, 6 women). Respiration of PBMC was measured and physical tests were performed in both groups twice with an interval of 6 months between measurements. Respiration of PLT was measured once. Blood was collected after overnight fasting and at least 48 h without intensive physical activity. Isolation of PBMC and PLT started 1 h after blood collection. PBMC were isolated in Ficoll-Pague™ PLUS gradient centrifugation medium using 50 mL Leucosep tubes . Whole blood and isolated blood cell fractions were counted with a Sysmex XN-350 hematology analyser. Bioenergetics of intact cells was determined at 37 °C in RPMI without glutamine in 2 mL chambers of the O2k-FluoRespirometer (Oroboros Instruments, Austria), applying a coupling control protocol extended by a respirometric cell viability test . Respiration of cells permeabilized with digitonin was measured in mitochondrial respiration medium MiR06 with addition of 20 mM creatine. Two different SUIT protocols were applied  with common cross-linked respiratory states, allowing for harmonization of both protocols. The fluxes were corrected for the contribution by contaminating cells.
Physical tests showed a strong difference between groups at both time points. VO2max in the active group was significanty higher than VO2max in the inactive group (42.1 ± 6.5 vs. 32.0 ± 6.4 mL·min-1·kg-1). The groups differed also in peak power output and maximal lactate accumulation at the end of the ergometric exercise test. In contrast to physical tests, respiration of PBMC and PLT did not differ between groups at any time point. The correction of PBMC respiration for the contribution by contaminating PLT decreased the variability of the measurements.
In summary, differences in lifestyle did not have a significant impact on respiration of blood cells when comparing an inactive with an active group of healthy subjects. The change to an increasingly passive lifestyle in patients with degenerative and neuromuscular diseases is expected to induce a decline of mitochondrial respiratory capacity measured in muscle biopsies. In contrast, our results suggest that the decline of bioenergetic capacity of PBMC in patients with deep depression  has diagnostic significance as an indicator of a specific pathological mitochondrial dysfunction and a functional marker of mitochondrial health.
Labels: MiParea: Respiration, Exercise physiology;nutrition;life style
Tissue;cell: Blood cells, Platelet Preparation: Intact cells, Permeabilized cells
Affiliations and support
- Sumbalova Z(1,2), Garcia-Souza LF(1,3), Menz V(3), Huber R(4), Burtscher M(3), Gnaiger E(1,4,5)
- Daniel Swarovski Research Lab, Dept Visceral, Transplant Thoracic Surgery, Medical Univ Innsbruck, Austria
- Pharmacobiochemical Lab, 3rd Dept Internal Medicine, Fac Medicine, Comenius Univ, Bratislava, Slovakia
- Inst Sport Science, Univ Innsbruck, Austria
- Sporttherapie Mag. Huber GmbH, Innsbruck, Austria
- Oroboros Instruments, Innsbruck, Austria. – firstname.lastname@example.org
- We thank Stephanie Droescher for technical assistance and Verena Laner for project administration. Contribution to K-Regio project K-Regio MitoFit funded in part by the Government of Tyrol within the program K-Regio of Standortagentur Tirol. Contribution to European Union Framework Programme Horizon 2020 COST Action CA15203 MitoEAGLE.
- Sumbalova Z, Hiller E, Chang S, Garcia-Souza LF, Droescher S, Calabria E, Volani C, Krumschnabel G, Gnaiger E (2016) Isolation of blood cells for HRR. Mitochondr Physiol Network 21.17:1-15. »»Bioblast Link
- Gnaiger E (2014) Mitochondrial pathways and respiratory control. An introduction to OXPHOS analysis. 4th ed. Mitochondr Physiol Network 19.12. Oroboros MiPNet Publications, Innsbruck:80 pp.
- Doerrier C, Sumbalova Z, Krumschnabel G, Hiller E, Gnaiger (2016) SUIT reference protocol for OXPHOS analysis by high-resolution respirometry. Mitochondr Physiol Network 21.06(01):1-12.
- Karabatsiakis A, Böck C, Salinas-Manrique J, Kolassa S, Calzia E, Dietrich DE, Kolassa IT (2014) Mitochondrial respiration in peripheral blood mononuclear cells correlates with depressive subsymptoms and severity of major depression. Transl Psychiatry 4:e397.