Abstract
There is a strong demand for diagnostic tools to identify athletes in various training states. PURPOSE: To determine if a cluster of proteins could be identified through proteomics procedures that are linked to functional overreaching (FOR) in male endurance athletes. METHODS: Participants (N=10, age 38.3±3.4 y, VO2max 41.3±1.7 ml.kg-1.min-1) served as their own controls and in random, counterbalanced order either ran/cycled 2.5 h (70.0±3.7% VO2max, 79.6±6.3% HRmax) three days in a row (FOR) or sat in the lab (rest) (separated by three weeks) (7:00 - 9:30 am, overnight fasted state). Participants provided fingerprick samples for dried blood spot samples (DBS) pre- and post-exercise/rest each of the three days, and then at 7:00 am during two additional recovery days. Participants also completed the Training Distress Scale (TDS) (19-items) at 7:00 am each of the five mornings during each trial (FOR and rest). Proteins were solubilized from DBS, digested into peptides and measured with nanoLC-MS in data independent acquisition mode (Q-Exactive, Thermo Fisher Scientific, Waltham, MA). The RAW MS data files were processed using SpectronautTM software (Biognosys, Schlieren, Switzerland). Following data independent acquisition (DIA method), 594 proteins were identified and quantified. Proteins were considered for the FOR cluster if they were elevated during one of the two recovery days but not more than one of the exercise days (compared to rest). The Generalized Estimating Equation (GEE) was used to identify proteins linked to FOR (between trial contrasts, P≤0.05 for proteins with CV<15%, P≤0.01 with CV>15%). RESULTS: TDS scores differed between FOR and rest trials, peaking on the first recovery day (9.8±3.8, 3.5±2.6, respectively, P=0.029). A total of 13 proteins was linked to FOR and of these, 11 were related to the immune system, and two to exercise-induced physiological responses. Immune-related proteins included those associated primarily with the acute phase response, complement activation, and granulocyte function. CONCLUSIONS: This study utilized targeted, DIA proteomics procedures to identify a cluster of 13 proteins linked to FOR (7.5 h of high intensity exercise over three days), and 85% of the proteins were related to immune system activation during the 2-day recovery period.
Highlights
Successful training leading to enhanced performance involves cycles of overload and adequate recovery [1,2,3]
No significant differences were found between the runners (N = 3) and cyclists (N = 7) for the data listed in Table 1 and exercise performance data, and all analyses were conducted for the combined group of these 10 athletes
Prior studies utilized a limited number of proteins, and most were based on acute shifts in racing dogs and horses
Summary
Successful training leading to enhanced performance involves cycles of overload and adequate recovery [1,2,3]. A primary goal during training is to avoid the combination of excessive overload and inadequate recovery leading to “overreaching”, defined as a short-term decrement in performance with or without related physiological and psychological symptoms in which restoration of performance takes several days to several weeks [1]. “Functional” overreaching occurs when athletes deliberately use a short-term period (e.g., training camp) to increase the training load resulting in short-term performance decrements without serious, long-lasting psychological or other negative symptoms [2]. Non-functional overreaching (NFOR or extreme overreaching) occurs when athletes train beyond their ability to recover with concomitant performance decrements and psychological disturbances that include decreased vigor and energy, increased fatigue, and loss of desire to train [1]. A hallmark feature of FOR and NFOR is the inability to sustain intense exercise for a prolonged period of time
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