Decrease in Cellular Nanovesicles Concentration in Blood of Athletes More Than 15 Hours After Marathon

Abstract

Zala Jan,1 Mitja Drab,2 Damjana Drobne,3 Apolonija Bedina Zavec,4 Mojca Benčina,5 Barbara Drasler,3 Matej Hočevar,6 Judita Lea Krek,1 Ljubiša Pađen,1 Manca Pajnič,1 Neža Repar,3 Boštjan Šimunič,7 Roman Štukelj,1 Veronika Kralj-Iglič1 1Laboratory of Clinical Biophysics, Faculty of Health Sciences, University of Ljubljana, Ljubljana, Slovenia; 2Laboratory of Physics, Faculty of Electrical Engineering, University of Ljubljana, Ljubljana, Slovenia; 3Nanobiology and Nanotoxicology Group, Biotechnical Faculty, University of Ljubljana, Ljubljana, Slovenia; 4Department of Molecular Biology and Nanobiotechnology, National Institute of Chemistry, Ljubljana, Slovenia; 5Department of Synthetic Biology and Immunology, National Institute of Chemistry, Ljubljana, Slovenia; 6Physics and Chemistry of Materials, Laboratory of Surface Engineering and Applied Surface Science, The Institute of Metals and Technology, Ljubljana, Slovenia; 7Institute for Kinesiology Research, Science and Research Centre Koper, Koper, SloveniaCorrespondence: Veronika Kralj-IgličLaboratory of Clinical Biophysics, Faculty of Health Sciences, University of Ljubljana, Ljubljana, SloveniaEmail [email protected]: Cellular nanovesicles (CNVs), that are shed from cells, have been recognized as promising indicators of health status. We analyzed the effect of long-distance running on concentration of CNVs, along with some standard blood parameters, in 27 athletes two days before and > 15 hours after physical effort.Methods: CNVs were isolated by repetitive centrifugation and washing of samples, and assessed by flow cytometry. Cholinesterase (ChE) and glutathione S-transferase (GST) activity were measured spectrophotometrically. Interleukin 6 (IL-6) and tumor necrosis factor-α (TNF-α) concentrations were measured using enzyme-linked immunosorbent assay (ELISA). C-reactive protein (CRP) was measured with immunoturbidimetric determination and lipidogram parameters were measured by enzymatic colorimetric assay. Flow cytometry was used for blood cell count and mean platelet volume (MPV) measurement.Results: More than 15 hours after physical effort a decrease was found in CNVs’ concentration in isolates from blood (46%; p< 0.05), in ChE activity in whole blood (47%; p< 0.001), in plasma (34%; p< 0.01), and in erythrocyte suspension (54%; p< 0.001), as well as in GST activity in erythrocyte suspension (16%; p< 0.01) and in IL-6 concentration in plasma (63%; p< 0.05). We found no change in GST activity in plasma and in TNF-α concentration in plasma. Correlations (> 0.8; p< 0.001) between CNVs’ concentration and ChE activity, and GST activity, respectively, in erythrocyte suspension were found.Conclusion: We found that > 15 hours post-physical effort, CNVs’ concentration was below the initial value, concomitant with other measured parameters: ChE and GST activity as well as IL-6 concentration, indicating a favorable effect of physical effort on health status. CNVs’ concentration and ChE activity in isolates from peripheral blood proved to have potential as indicators of the response of the human body to inflammation after physical effort. Physical activity should be considered as an important factor in preparation of subjects for blood sampling in procedures focusing on CNV-containing diagnostic and therapeutic compounds.Keywords: membrane vesiculation, physical effort, blood samples, inflammation process, cellular nanovesicles, marathon

Keywords

• membrane vesiculation
• physical effort
• blood samples
• inflammation process
• cellular nanovesicles
• marathon