Home > Journals > Medicina dello Sport > Past Issues > Medicina dello Sport 2018 June;71(2) > Medicina dello Sport 2018 June;71(2):203-15

CURRENT ISSUE
 

JOURNAL TOOLS

eTOC
To subscribe
Submit an article
Recommend to your librarian
 

ARTICLE TOOLS

Publication history
Reprints
Permissions
Cite this article as

 

MEDICAL AREA   

Medicina dello Sport 2018 June;71(2):203-15

DOI: 10.23736/S0025-7826.18.03314-8

Copyright © 2018 EDIZIONI MINERVA MEDICA

language: English, Italian

Analysis of the modifications of erythrocyte membrane proteome induced by blood storage

Chiara FOSSATI 1 , Loredana GRASSO 1, Barbara PERGOLIZZI 2, Giuliana ABBADESSA 2, Silvia RACCA 2, Alessandro SALUTO 3, Odetta CAMERINI 3, Fabio PIGOZZI 1, Paolo BORRIONE 1

1 Department of Movement, Human and Health Sciences, University of Rome “Foro Italico”, Rome, Italy; 2 Department of Clinical and Biological Sciences, University of Turin School of Medicine, San Luigi Gonzaga University Hospital, Orbassano, Turin, Italy; 3 Department of Blood Transfusion, San Luigi Gonzaga University Hospital, Orbassano, Turin, Italy


PDF


BACKGROUND: There is currently no direct method to detect the use of autologous blood transfusions in the context of anti-doping controls. The storage of erythrocytes causes changes in membrane proteins, which could be evidenced by proteomics. The aim of the present study was to identify these modifications on blood samples stored in transfusion bags, at different storage time points.
METHODS: Twenty-three hemochromatosis patients undergoing periodic phlebotomy sessions for therapeutic purposes at the San Luigi Gonzaga Hospital in Orbassano, Turin, were enrolled in the study. The blood was stored under standard conditions for up to 21 days at the local Blood Bank. The samples were analyzed at 0, 7, 14, and 21 days after phlebotomy, in order to isolate the erythrocyte membranes. After identifying the most valid erythrocyte membrane extraction protocol and obtaining homogeneous protein samples by manual sonication, we performed Western blot assay (WB), by incubating the membranes with the primary anti β-actin antibody and then with the anti-band 3 antibody. Two-dimensional electrophoresis was then performed on the samples with the aim of highlighting different protein patterns, that were later identified by mass spectrometry (MALDI-TOF).
RESULTS: An increase in actin expression over time was evidenced by WB. Two-dimensional electrophoresis, followed by MALDI-TOF analysis, showed an increase in the number of actin spots and its CRA-b isoform from day 0 to day 21. The analysis of band 3 showed a gradual fragmentation of the protein during storage time.
CONCLUSIONS: The results of the present study confirmed the observations of scientific literature demonstrating a modification of erythrocyte proteins during red blood cells (RBC) storage. However, further investigations are needed in order to univocally define these modifications and to make the results applicable in the context of the strategies to fight doping.


KEY WORDS: Proteomics - Erythrocytes - Autologous blood transfusions - Doping in sports

top of page