TABLE 3.
Studies investigating the protein, protein/nucleic acid or metabolite cargo of exercise‐induced EVs.
| EV isolation | EV analysis | Type of exercise | Time of EV collection | EV source | Health status | Cohort | Gender | Molecular changes | EV term | References |
|---|---|---|---|---|---|---|---|---|---|---|
| Sequential centrifugation, max. speed 20,000 × g | NTA, EM | 1‐h bout of cycle ergometer exercise | Before, immediately after, and 4 h after exercise | Plasma | Healthy | 11 (mean age 27, SEM 1) | Male | 322 proteins were altered immediately after exercise. 3 proteins (Vascular non‐inflammatory molecule 2, S100‐A8, S100‐A9) were significantly different between rest and recovery samples | EVs | Whitham et al. (2018) |
| Acoustic trapping and sequential centrifugation max. speed 13,000 × g or 20,000 × g | FC, EM, NTA | 10 min workout with progressively increasing intensity on stationary bicycles | Before and 1 h after exercise | Plasma | Healthy | 20 (mean age 58) | not specified | Cardiovascular disease panels CVD II and III: 54 proteins were changed in response to physical exercise including chemokines and interleukins associated with the inflammatory response (e.g. CCL17, CCL24, CXCL1, CXCL16, IL1RA, IL18, and macrophage receptor with collagenous structure (MARCO)), as well as proteins associated with angiogenesis (e.g. ANG1 and TIE2) and coagulation (PAR1, SELP, SRC, and vWF) | EVs | Bryl‐Górecka et al. (2018) |
| SEC and immuno‐isolation | NTA, WB, EM, FC | Incremental cycling test until exhaustion | Before, during and immediately after exercise | Plasma | Healthy | 21 active (mean age 28.7 ± 4.2) | Male | EVs release after exercise contain the lymphocyte markers CD4 and CD8, the monocyte marker CD14, the platelet markers CD41, CD42 and CD62P, the endothelial markers CD105 and CD146 and the antigen presenting cell marker MHC‐II | EVs | Brahmer et al. (2019) |
| EV Array and polymer‐based precipitation | NTA, TEM, WB | Single bout of blood flow restricted resistance exercise consisting of 5 sets of knee extensions | Immediately before exercise and after 1 h | Plasma | Healthy | 6 recreationally active (age 21 ± 0.6) | Male |
The proteins CD41, NCAM, Alix and CD25 were upregulated and Flotillin‐1 was down‐regulated after exercise. The miRNAs miR‐182‐5p, miR‐1294, let‐7b‐5p, miR‐451a, miR‐16‐5p, and miR‐36‐3p were upregulated and miR‐19b‐3p, miR‐17‐5p, miR‐221‐3p, miR‐150‐5p, miR‐340‐5p and miR‐21‐5p were downregulated after exercise |
EVs | Just et al. (2020) |
| Sequential centrifugation, max. speed 100,000 × g | WB, NTA, TEM | 16 resistance sessions over 8 weeks (2 per week). 8 different resistance exercises, with three sets | Before and after training | Plasma | Healthy | 28 trained (age 73.6 ± 0.8) and 10 control (age 72.6 ± 0.4) | Male, Female | The percent change of CD63 was reduced in the trained group versus the control group after an 8‐week resistance training program | Exosomes | Estébanez et al. (2021) |
| SEC | NTA, TEM, WB | A progressive 12‐week supervised cycle ergometer training of 35 min. Exercise sessions were repeated 3 times weekly on non‐consecutive days | At baseline and 72 h after the last bout of the 12‐week period training | Serum | Healthy or type 2 diabetes | 5 type 2 diabetes, 5 insulin resistant nondiabetic, 5 insulin sensitive nondiabetic (age 55–59) | Male | 262 proteins were regulated (102 down‐regulated, 160 up‐regulated) by exercise, including ALIX, Rab proteins annexins, flotillin, moesin, tubulin, integrins. 13 of the 262 proteins were shared between the 3 groups including antithrombin III, kininogen I, histidine‐rich glycoprotein, and α1‐antitrypsin. 29 proteins, including the fibrinogens FGA, FGB, and FG, were up‐ or down‐regulated after exercise only in the 2 insulin‐resistant groups | Exosomessmall EVs | Apostolopoulou et al. (2021) |
| SEC | FC, NTA, WB | 4 min of high‐intensity interval training in a programmable bicycle ergometer consisting of 8 sets of bout (20 s) and rest (10 s) | Before, at the immediate conclusion, 30 min and 120 min after exercise | Serum | Healthy | Total cohort: 17 (median age 20). For proteomics 3 men were selected | Male | 20 proteins were changed when comparing before and after exercise: alpha‐2‐antiplasmin, fibronectin, peroxiredoxin‐2, von Willebrand factor, multimerin‐1, fibrinogen alpha chain, fibrinogen beta chain, fibrinogen gamma chain, tetraspanin 9, spectrin beta chain, cofilin‐1, carbonic anhydrase‐1, catalase, immunoglobulin heavy variable 1–69, cholinesterase, immunoglobulin heavy constant gamma 4, histidine‐rich glycoproteins, serotransferrin, complement factor H and haptoglobin | EVs | Kobayashi et al. (2021) |
| 10,000 × g centrifugation, followed by SEC and UC at 100,000 × g for 1.5 h | NTA, EM, Nanoscale flow cytometry, mass spectrometry |
Aerobic exercise: one bout of high‐intensity cycling. Resistance exercise: one moderate bout of single leg exercise |
Aerobic exercise: before, 10 min, 1 h and 3 h after exercise Resistance exercise: before, 10 min, 30 min and 60 min after exercise. |
Plasma | Healthy |
Aerobic exercise: 7 male (mean 37.0 ± 5.0), 7 female (mean 28.0 ± 4.4) Resistance exercise: 8 male (mean 32.6 ± 5.5), 8 female (mean 33.7 ± 10.2) |
Males, females |
10 min after aerobic exercise:321 proteins increased (Ingenuity Pathway Analysis: immune regulation, glycolysis, signaling via Rho GTPase, interleukin‐8, and integrin) and after resistance exercise 7 proteins up‐ and 1 protein downregulated. Common to both: MMP9, MPO3, HIST1H4A |
EVs | Vanderboom et al. (2021) |
| miRCURY exosome cell/urine/CSF Kit | Exoview | 20 m shuttle run test | Before, immediately after, and after 1 h rest. | Urine | Healthy | 13 (age 25.5 ± 2.0) | Male | CD9, CD63, and CD81 increased in post‐exercise. 1 h after exercise only CD9 was still increased. miR‐let‐7a‐5p, miR‐193b‐3p, miR‐23a‐3p, miR‐320a‐3p, miR‐423‐5p, miR‐4454, miR‐5100, miR‐7847‐3p and miR‐8485 were altered after exercise | EVs | Park and Moon (2022) |
| Sequential centrifugation, max. 100,000 × g | Spectradyne nCS1, WB | 20 min of moderate intensity cycling | Before and after exercise | Plasma | Healthy | 20 young (18–35) and 20 mature (50–70) fit and unfit | Male | NAMPT (nicotinamide phosphoribosyltransferase) is increased after exercise in young individuals with high aerobic fitness whereas mature fit and young unfit individuals exhibited a limited increase | EVs | Chong et al. (2022) |
| Sequential centrifugation, max. 110,000 × g; SEC | NTA, WB, FC | Four different exercise protocols: Acute Aerobic Exercise (AAE) and Aerobic Training (AT), Acute Maximal Aerobic Exercise (AMAE), Altitude Aerobic Training (AAT) | AAE and AT: before and after 0,1, 2, 6, and 24 h. AMAE: 1 h before, and after 0, 1 and 2 h. AAT: before and at the end of the training, at least at 48 h from exercise. | Plasma | Healthy | 13 not physically active (age 20.1 ± 0.6) and 6 male athletes (experience in the distance 5000–10.000 m of 5 ± 1 years) men (age 23.3 ± 6.8) | Male, Female | miR‐206, miR‐133b and miR‐146a were upregulated following AAE within the first hour after exercise in the sedentary group. miR‐146 is increased immediately post‐exercise in the AT group. miR‐206, miR‐133b, miR‐486‐5p, miR‐181a‐5p and miR‐16 are upregulated immediately after AMAE. AAT showed no statistically significant variations following the training regarding miRNA expression levels. CD42a, CD41b, CD29, CD40 and CD62P are increased at 1 h post‐exercise and CD56, CD105 and CD3 are detected only immediately after AAE. mtDNA (COX1) is increased 1 h after AAE | EVs | Maggio et al. (2023) |
| Immuno‐capture/ imaging system | Exoview | High intensity intermittent cycling: 4 × 30 s at 200% of individual max power | Before and immediately after exercise. | Plasma | Healthy | 7 recreationally active persons (age 21 ± 4) | Male, Female | The expression of CD9 and CD63 is increased after high intensity intermittent exercise | small EVs | McIlvenna et al. (2023) |
| Filtration followed by centrifugation at 100,000 × g | BCA protein assay, WB, NTA, EM | A cycling bout exercise starting with an intensity of 40 W and increasing resistance with 15 W for females and 20 W for males every 2 min until exhaustion | Sweat was collected during exercise and during a rest period of 30 min. | Sweat | Healthy | 10 (3 females age 33.7 ± 6.6; 7 males age 38.7 ± 3.6) | Male, Female | Glutamate and glutamine (glutamate metabolism pathway), alanine, arginine, glycine, proline, threonine, and serine (amino acid pathway) and lactate (glycolysis pathway) were significantly increased during the exercise test compared to during recovery. Lysine was only detected during the exercise test and not during recovery. Myristate (fatty acid pathway) was reduced during exercise compared to during recovery. | EVs | Ali et al. (2023) |
| ExoQuick, immuno‐captured with L1CAM | NTA, FC | Supervised moderate to high‐intensity aerobic exercise of 60‐min duration 3 times a week for 16 weeks | At baseline and after 16 weeks of training. | Plasma | Mild to moderate Alzheimer's disease | 48 (age 71.3 ± 6.5) in exercise group and 47 (age 71.2 ± 6.5) in control group | Male, Female | The neuroprotective proteins proBDNF, BDNF, and humanin are increased in the exercise but not in the control group | Neuron‐derived EVs | Delgado‐Peraza et al. (2023) |
| SEC | NTA, ImageStream, Bradford colorimetric assay, WB, EM |
Endurance training allowing men to have specific running or strength performance |
At rest (circa 40 h after the most recent exercise session) on two separate but identical lab visits | Plasma | Healthy | 13 endurance‐trained (age 30 ± 6), 13 strength‐trained (age 25 ± 5) and 12 recreationally active (age 26 ± 2) | Male | 96 metabolites are unchanged when comparing resting men with divergent histories of exercise training | small EVs | Darragh et al. (2023) |
| SEC | NTA, WB, EM, Surface plasmon resonance imaging | Single bout of endurance exercise consisting of 30 min exercise on a treadmill | At basal level and immediately after exercise | Plasma | Healthy | 21 (age 26.95 ± 3.07) with a medium/high fitness level | Male | 98 proteins were upregulated after exercise including MAP2K1, and carnosine dipeptidase 2, glutamate‐cysteine ligase modifier subunit and microsomal glutathione S‐transferase 2, which are essential for the glutathione biosynthetic process. 48 of the proteins have catalytic activity, and the most upregulated proteins are mitochondrial proteins involved in the maintenance of energetic balance. The activity of Glutathione Reductase and Catalase (antioxidant enzymes) was higher after exercise | EVs | Lisi et al. (2023) |
| ExoQuick | NTA, protein assay kit, WB | A single 30 min bout or 5 × 30 min on consecutive days of aerobic exercise in a treadmill with a specific speed and slope | Before, 3 and 24 h after a bout of exercise and after 24 h from the last session of a 5‐day consecutive training. | Plasma | Healthy | 19 (age 25 ± 3.2) divided into a trained (Schmidt et al., 2015) and an untrained (Li et al., 2022) group | Male | Protein carbonyl, catalase, SOD2, and HSF1 were higher at baseline in untrained compared to trained men, and SOD2 was also increased after 3 h. However, Tot‐HSP27 was higher in the trained group at each time point. Protein carbonyl, HSF1, Catalase, and SOD2 were lower after 5 days of regular aerobic compared with the same group before training and the untrained group | EVs | Lisi et al. (2023) |
| SEC followed by centrifugation at 120,000 × g | NTA, WB, EM, immuno‐capture | 12 week of home‐based progressive elastic band‐based resistance training 3 days a week | Prior to and following a 12‐week exercise program | Plasma | Healthy | 30 (age 74.9 ± 5.7) | not specified | TSG101 and the miRNAs miR‐23a, miR‐27a, miRNA‐199a, miR‐146a and miR‐92a increased after exercise | Exosome‐like vesicles | Xhuti et al. (2023) |
| SEC | NTA, EM | Stationary bicycle exercise with an increase in workload of 15 watts per minute until volitional exhaustion | At baseline immediately prior to exercise, and 15 min and 24 h after exercise. | Plasma | Healthy or myalgic encephalomyelitis/ chronic fatigue syndrome (ME/ CFS) | 18 (age 44.4 ± 7.9) patients and 17 sedentary (age 44.2 ± 13.4) | Female |
MYL9, PDIA6, PPIB, HSPA5, F8, HSP90B1, VDAC3, FN1, CANX, CLTC, F13A1 are less abundant and ANXA2, B2M, ORM1 are more abundant in the ME/CFS group 15 min after exercise. EMILIN is more abundant in ME/CFS patients 24 h after exercise. 63 proteins (all upregulated, including YWHAE, YWHAQ, YWHAB, YWHAZ, YWHAG, YWHAH, ITGB1, ITGB3 and ITGA2B) in the ME/CFS group and 187 proteins (178 upregulated including CCT2, CCT5, CCT6A, CCT8, TUBB, TUBB4B, TUBA4A, TUBA8, MYL6, MYL9, MYL12A, MYLK, TPM4, TMOD3) and 9 ‐AGT, C4B, CLU, COLEC11, FCN2, MBL2, ORM2, PON1 and VSIG6‐ downregulated) in the control group were found differentially expressed after 15 min exercise |
EVs | Giloteaux et al. (2024) |
| Sequential centrifugation at max. speed 100,000 × g | Particle analysis with Spectradyne nCS1 and WB | A single 20‐min boat of moderate‐to‐vigorous intensity cycling | Before and immediately after exercise | Plasma | Healthy | 14 young (age 27.1 ± 4.0) and 14 mature (age 60.9 ± 6.1), 14 unfit (age 44.4 ± 18.7), 14 fit (age 43.7 ± 17.8) | Male | ADAMTS13, ANXA1, ANXA6, BASP1, CTSG, F8, HIST1H4A, ITGB2, LCP1, LTF, LYZ, MPO, and S100A8 are increased after exercise in the total cohort. ANXA1, LCP1, LYZ, MPO, and S100A8 are upregulated after exercise in the young subset. ANXA1, BASP1, HIST1H4A, LTF, LYZ, MPO, S100A8, and S100A9 are upregulated after exercise in the mature subset. ANXA1, LYZ, S100A8, and S100A9 were upregulated in the unfit subset after exercise. ANXA1, ANXA6, EEF1A1, HIST1H4A, ITGB2, LCP1, LTF, LYZ, MPO, and S100A8 are upregulated in the fit subset | small EVs | Chong et al. (2024) |
Abbreviations: BCA, bicinchoninic acid; EM, electron microscopy; FC, flow cytometry; NTA, nanoparticle tracking analysis; SEC, size‐exclusion chromatography; WB, Western blot.