Circulating microRNA levels after exercise-induced muscle damage and the repeated bout effect.
Author(s): Chalchat E, Martin V, Charlot K, Bourrilhon C, Baugé S, Bourdon S, Gruel A, Lepetit B, Banzet S, Garcia-Vicencio S, Siracusa J
Publication: Am J Physiol Regul Integr Comp Physiol, 2022, Vol. , Page
PubMed ID: 36374177 PubMed Review Paper? No
Purpose of Paper
The purpose of this paper was to compare muscular maximal voluntary contraction (MVC) torque; serum creatinine kinase (CK) activity and levels of myoglobin; and plasma levels of muscle-specific microRNAs (miRNA, miR) immediately (0 h), 2, 6, 24 and 48 h after downhill walking; data was collected after a second bout of exercise two weeks after the first. . The ability of CK activity and levels of myoglobin and miRNAs to predict a decrease in MVC torque >24 h post-exercise was also investigated.
Conclusion of Paper
Levels of maximal voluntary contraction (MVC) torque deficit, serum creatinine kinase (CK), and myoglobin, and muscle-specific miRNAs in plasma were comparable when quantified prior to each bout of exercise. MVC torque was significantly lower than baseline (quantified immediately before exercise) 24 h and 48 h after the first bout of exercise, but did not differ from baseline after the second bout of exercise. While increased muscle soreness occurred 24 and 48 h after both bouts of exercise, soreness was significantly lower following the second bout of exercise compared to the first. Levels of miR-1-3p, miR-133a-3p, miR133b, and miR-206 significantly increased after exercise, but significance was dependent on bout (first or second), the miRNA targeted, and timepoint (2, 6, 24, or 48 h after exercise). Creatinine kinase levels were higher than pre-exercise levels 2, 6, 24 and 48 h post-exercise (both bouts) and myoglobin levels were higher than pre-exercise levels immediately following, 2 h, and 6 h post-exercise (both bouts). Levels of CK activity were significantly correlated with miR-1-3p and miR-133a-3p levels, and levels of myoglobin were correlated with miR-1-3p, miR-133a-3p and miR-206 levels. The best markers to predict a decrease in MVC torque >24 h post-exercise were a combination of myoglobin, CK activity, miR-1-3p, miR-133b and miR-206 levels when markers were quantified immediately following exercise; while a combination of myoglobin and CK activity were the best predictors of a decrease in MVC torque when quantification occurred 2 h post-exercise; and myoglobin and miR-1-3p levels were the best predictors when samples were collected 6 h post-exercise.
Studies
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Study Purpose
The purpose of this study was to compare maximal voluntary contraction (MVC) torque; and serum levels of CK activity, and myoglobin, and plasma levels of muscle-specific miRNAs 0, 2, 6, 24 and 48 h following two bouts of downhill walking (bouts occurred two weeks apart). The ability of CK activity and levels of myoglobin and miRNAs to predict a decrease in MVC torque >24 h post-exercise was also investigated. Blood was collected from nineteen men (aged 31.3 ± 5.6 years, body mass index 24.0 ± 2.3 kg/m2) who regularly exercised (3-8 h/week) and had no history of muscular, joint, or bone disorders. All participants had not participated in strenuous mountain trekking, downhill walking or resistance training for at least six months prior to the study. Blood was collected before and 0, 2, 6, 24 and 48 h after downhill walking on a treadmill for 45 min wearing a weighted vest (10% of body mass) and again two weeks later before and 0, 2, 6, 24 and 48 h after downhill walking on a treadmill for 45 min wearing a backpack (20% of body mass). Blood was collected into EDTA and serum separator tubes (SST). After clotting, both tubes were centrifuged at 2,000 g at 4°C for 10 min and serum and plasma were aliquoted and stored at -80°C. Serum CK activity and myoglobin levels were analyzed on an autoanalyzer. RNA was extracted from plasma using the mirVana PARIS Kit, stored at -80°C, and reverse transcribed with the miRCURY LNA RT Kit. Levels of muscle-specific miRNAs (miR-1-3p, miR-133a-3p, miR-133b, miR-206, miR-208a-3p, miR-208b-3p, miR-378a-3p, and miR-499a-5p) were quantified by real-time PCR. MVC torque was measured pre-exercise, 24 h and 48 h post exercise using an isokinetic dynamometer. Muscle soreness was rated on a 100 mm line scale while performing a squat.
Summary of Findings:
Levels of maximal voluntary contraction (MVC) torque, serum creatinine kinase (CK) and myoglobin, and muscle specific miRNAs in plasma were comparable when quantified before each bout of exercise. MVC torque was significantly lower 24 h and 48 h after the first bout of exercise compared to pre-exercise levels (baseline)(P<0.001), but did not differ from baseline after the second exercise bout; thus,MVC torque was lower 24 and 48 h after the first bout of exercise when compared to the second bout (P<0.01, both). While increased muscle soreness occurred 24 and 48 h after both bouts of exercise (P<0.001, all), soreness was significantly lower following the second bout of exercise compared to the first for both timepoints (P<0.001, both). Levels of miR-1-3p, miR-133a-3p, miR133b, and miR-206 significantly increased after exercise, while levels of miR-208a-3p, miR-208b-3p, miR-378-3p, and miR-499a-5p were unaffected. Levels of miR-1-3p, miR-133a-3p, miR133b, and miR-206 were significantly elevated 2 h and 6 h after the first bout of exercise (P<0.05,all), miR-206 was elevated immediately following the second bout of exercise (P<0.05), miR-1-3p and miR-133b were elevated 2 h after the second bout of exercise (P<.05, both), and miR-133b and miR-133a-3p were elevated 6 h following the second bout of exercise (P<0.05 and P<0.01, respectively). Importantly, significant differences in levels of miR-1-3p and miR-133a-3p were observed between exercise bouts when specimens were collected 6 h post-exercise (P<0.05, both). Creatinine kinase activity levels were higher 2, 6, 24 and 48 h post-exercise than pre-exercise levels for both bout one (P<0.05, P<0.001, P<0.001, and P<0.001, respectively) and two (P<0.01, P<0.001, P<0.001, P<0.05, respectively), with the highest levels observed 24 h post-exercise (both bouts). Myoglobin levels were higher immediately following (P<0.01 both bouts), 2 h (P<0.001 both bouts), and 6 h (P<0.001 both bouts) post-exercise compared to pre-exercise levels. CK activity was significantly correlated with miR-1-3p level (ρ=0.54, P<0.001); and miR-133a-3p (ρ=0.58, P<0.001) and myoglobin levels were significantly correlated with levels of miR-1-3p (ρ=0.50, P=0.005), miR-133a-3p (ρ=0.36, P=0.048), and miR206 (ρ=0.48, P=0.007). A decline in MVC torque was negatively correlated with myoglobin levels 2 h post-exercise (ρ=-0.5159, P=0.0035), and with CK activity, myoglobin levels, and miR-1-3p and miR-206 levels 6 h post-exercise (ρ=-0.4295, P=0.0071; ρ=-0.5315, P=0.0025; ρ=-0.3831, P=0.0176; and ρ=-0.3767, P=0.0197, respectively). The best markers to predict a decline in MVC torque >24 h post-exercise were a combination of myoglobin level, CK activity, and miR-1-3p, miR-133b and miR-206 levels quantified immediately following exercise; a combination of myoglobin level and CK activity were the best predictors when quantification occurred 2 h post-exercise; and myoglobin and miR-1-3p levels were the best predictors when quantified 6 h post-exercise.
Biospecimens
Preservative Types
- Frozen
Diagnoses:
- Not specified
Platform:
Analyte Technology Platform Protein Clinical chemistry/auto analyzer RNA Real-time qRT-PCR Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Acquisition Time of biospecimen collection Pre-exercise
Immediately post-exercise
6 h post-exercise
24 h post-exercise
48 h post-exercise
2 h post-exercise