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Effects of ultramarathon performance on mitochondrial respiration in human platelets - a project summary

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Revision as of 09:37, 28 October 2015 by Hoppel Florian (talk | contribs)

See also: Hoppel 2015 Abstract MiP2015


Introduction

Acute strenuous exercise is linked to severe inflammatory responses [1,2], alterations of mitochondrial function of human skeletal muscle and increased oxidative stress [3]. Whereas mitochondrial function in human platelets and lymphocytes has been characterized in various disease states, little is known about potential effects of strenous exercise on respiration and production of reactive oxygen species (ROX). Importantly, respiratory capacity of human Peripheral Blood Mononuclear Cells (PBMCs: lymphocytes and monozytes) was linked to physical fitness [4], supporting the concept that mitochondrial function in human blood cells can be used as a systemic mitochondrial marker. Therefore, in combination with eased sampling compared to invasive muscle biopsies, human blood cells seem to gain significance in mitochondrial science. To our knowledge, there is just one existing paper examining influence of acute intensive exercise on respiration of human blood cells [2], we investigated the influence of completion of an ultramarathon on mitochondrial respiration in human platelets.


Methods

10 male hobby athlethes were chosen for the study and signed a written conset. 3 blood samples a 18 ml (EDTA) were taken to highlight potential influence of ultramarathon performance and time of recovery on human platelets: ~10 h before competition start (PRE), ~15 min after finishing (POST). Whole-blood samples were centrifuged immediately (15 min, 300 g) to gain platelet-rich plasma, which was incubated 20 min before performing the second centrifugation step (1000 g, 10 min). Thus, pelleted platelets were resuspended in MiR05 by gentle pipetting and added to the OROBOROS O2k. Cells were permeabilized to ensure access of external substrates. Following SUIT was performed:


substrate P M Dig ADP G S u Rot Ama
conc. 5 mM 2 mM 200 µg/ml 1 mM 5 mM 10 mM 1 µM doses up to Jmax 2 µM 1.25 µM

Table 1: P: Pyruvate, M: Malate; Dig: Digitonine, G: Glutamate, S: Succinate; u: uncoupler (CCCP: carbonyl cyanide m-chloro phenyl hydrazone); Rot: Rotenone; Ama: Antimycin A.


To ensure earliest possible sample analysis, 6 O2k were running simultaneously in the locally set-up laboratory in the municipal building of Gmunden close to the start/finish area. After respiratory analysis, samples were extracted from the O2k-Chambers and frozen for further analysis of protein content. Additional blood was taken to perform further analysis (oxidative stress markers, lymphocyte/monocyte/neutrophils content, creatin kinase, lactate dehydrogenase, creatinine).


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Pic. 1: sample preparation in our "field laboratory"


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Pic. 2 & 3: platelet separation by centrifugation


Competition and participants

To enhance the chance for possible effects, a rather challenging competition with a total distance of 68 kilometers and approximately 4500 m of ascent was chosen. The course was set around the lake "Traunsee" by crossing all peaks around it, whereby the participants had the opportunity to choose their start time unfettered between 3 and 5 am in Gmunden/Austria.

Being hobby athletes, participants exhibited a mean BMI of 24.9 kg/m² and an average age of 39.9 years. The average speed of 5 km/h and a heart rate of 134 bpm during the race goes along with an average finish time of 12.9 h, whereas individual finish times were rather divergent (min: 9.7 h; max: 16.1 h).


Course.jpeg

Pic. 4: competition course


Start.jpg

Pic. 5: start during the night


Läufer.jpg

Pic. 6: somewhere on the course


Preliminary results of platelet respiration


Discussion


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Pic. 7: 2 O2ks at work


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Pic. 8: cellcounting with the Casy model TT


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Pic. 9: first discussion of results


References


1. de Gonzalo-Calvo D, Dávalos A, Montero A, García-González A, Tyshkovska I, González-Medina A, Soares S, Martínez-Camblor P, Casas-Agustench P, Rabadán M, Díaz-Martínez AE, Úbeda N, Iglesias-Gutiérrez E (2015) Circulating inflammatory miRNA signature in response to different doses of aerobic exercise. J Appl Physiol 2:124-34.

2. de Lucas DR, Caputo F, Mendes de Souza K, Sigwalt AR, Ghisoni K, Lock Silveira PC, Remor AP, da Luz Scheffer D, Antonacci Guglielmo LG, Latini A (2014) Increased platelet oxidative metabolism, blood oxidative stress and neopterin levels after ultraendurance exercise. J Sports Sci 32:22-30.

3. Tonkonogi M, Walsh B, Svensson M, Sahlin K (2000) Mitochondrial function and antioxidative defence in human muscle: effects of endurance training and oxidative stress. J Physiol 528:379-88.

4. Tyrrell DJ, Bharadwaj MS, Van Horn CG, Marsh AP, Nicklas BJ, Molina A (2015) Blood-cell bioenergetics are associated with physical function and inflammation in overweight/obese older adults. Exp Gerontol, doi: 10.1016/j.exger.2015.07.015.