Purpose Previous studies have suggested that circulating extracellular vesicles (EVs) arise after high intensity exercise and urine could reflect the plasma proteome. Herein, we investigated the characteristic of urinary EVs from healthy young adult males who had completed a maximal effort exercise test. Methods Thirteen healthy men completed a 20 m shuttle run test (20 m SRT). Fresh urine samples were collected at first morning, right after, and 1 h rest after 20 m SRT. Also, blood lactate, heart rate, rating of perceived exertion, and blood pressure were measured before, right after, and 1 h rest after 20 m SRT. Urinary EVs were analyzed using Exoview instrument and microRNAs (miRNAs) sequencing on urinary EVs were performed. Results Urinary EVs increased significantly after exercise and returned to baseline value after 1 h of rest. miRNA sequencing on urinary EV revealed alterations in four miRNAs (1 up and 3 down) and nine miRNAs (2 up and 7 down) in pre- vs. post- and post- vs. post-1 h samples, respectively. Lastly, bioinformatic analysis of urinary EV miRNA suggests that predicted target genes could affect PI3K-Akt, mitogen-activated protein kinase, and insulin pathways by exercise. Conclusions Exercise to voluntary exhaustion increased the number of EVs in urine. Also, miRNAs in urinary EVs were altered after exercise. These findings could indicate the possibility of using the urinary EVs as a novel biomarker of acute exercise-induced fatigue.
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