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Sumbalova 2017 MiP2017

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Revision as of 07:48, 20 October 2017 by Kandolf Georg (talk | contribs)
Zuzana Sumbalova
The effect of lifestyle on respiration of peripheral blood mononuclear cells and platelets.

Link: MiP2017

Sumbalova Z, Garcia-Souza LF, Menz V, Burtscher M, Gnaiger E (2017)

Event: MiP2017

COST Action MITOEAGLE

Blood cells are an attractive source of mitochondria that could be used in human studies for diagnostic purposes. The aim of our study was to find out whether regular physical activity can affect respiration of isolated peripheral blood mononuclear cells (PBMC) or platelets (PLT).

A group of 15 healthy participants (10 men, 5 women) aged 39-64 years with active lifestyle, participating in regular supervised training once a week for several years (active group) was matched with a group of participants without any regular physical activity for several years (non-active group, 9 men and 6 women). Respiration of PBMC was measured and physical tests were performed in both groups twice with the interval of 6 months between the measurements. Respiration of PLT was measured once. Blood was collected after overnight fasting and at least 48 hours without intensive physical activity. Isolation of PBMC and PLT started 1 hour after blood collection. PBMC were isolated on Ficoll-Pague™ PLUS gradient centrifugation medium using 50 mL Leucosep tubes [1]. Whole blood as well as isolated blood cell fractions were counted on Sysmex XN-350 hematology analyzer. Bioenergetics of intact cells was determined at 37 °C in RPMI without glutamine in 2 mL chambers of O2k-FluoRespirometer (Oroboros Instruments, Austria) applying a Coupling Control Protocol extended for cell membrane permeability tests [2]. For evaluation of mitochondrial respiration, mitochondrial respiration medium MiR06 with addition of 20 mM creatine was used. After permeabilization of cells with digitonin, two different SUIT protocols were applied [3] with common cross-linked respiratory states, allowing harmonization of both protocols. The fluxes were corrected for contribution by contaminating cells.

Physical tests showed a strong difference between groups at both time points. VO2 max in the active group was significanty different from VO2 max in the active group (42.1 ± 6.5 mL·min-1·kg-1 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 exercise test. In contrast to physical tests, respiration of PBMC or PLT did not differ between groups at any time point. The correction of PBMC respiration for contribution by contaminating PLT decreased variability of the measurements.

In summary we can conclude that active lifestyle did not have impact on respiration of blood cells. Therefore we suppose that lifestyle will most probably not interfere with diagnostics of mitochondrial dysfunction in pathological states when using blood cell respiration as a functional marker for mitochondrial health. In evaluation of respiratory measurements, the degree of contamination of cell preparation should be taken into account.


Bioblast editor: Kandolf G


Labels: MiParea: Respiration, Exercise physiology;nutrition;life style 


Tissue;cell: Blood cells, Platelet  Preparation: Intact cells, Permeabilized cells 



HRR: O2k-FluoRespirometer"O2k-FluoRespirometer" is not in the list (Oxygraph-2k, TIP2k, O2k-Fluorometer, pH, NO, TPP, Ca, O2k-Spectrophotometer, O2k-Manual, O2k-Protocol, ...) of allowed values for the "Instrument and method" property. 


Affiliations

Sumbalova Z(1,2), Garcia-Souza LF(1,3), Menz V(3), Burtscher M(3), Gnaiger E(1,4)
  1. Daniel Swarovski Research Lab, Dept Visceral, Transplant Thoracic Surgery, Medical Univ Innsbruck, Austria
  2. Pharmacobiochemical Lab, 3rd Dept Internal Medicine, Fac Medicine, Comenius Univ, Bratislava, Slovakia
  3. Inst Sport Science, Univ Innsbruck, Austria
  4. Oroboros Instruments, Innsbruck, Austria. – zuzana.sumbalova@i-med.ac.at

References and support

  1. 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
  2. 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.
  3. 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.
We thank Stephanie Droescher for technical assistance and Verena Laner for project administration. Contribution to K-Regio project MitoFit funded in part by the Government of Tyrol within the program K-Regio of Standortagentur Tirol. Contribution to COST Action CA15203 MitoEAGLE.