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. 2025 Jul 17:16:1583870.
doi: 10.3389/fphys.2025.1583870. eCollection 2025.

Maximal and sub-maximal exercise tests alter PBMC microRNA expression: insights into sport- and sex-specific variations

Guy Shalmon  1   2 , Guy Shapira  2   3 , Rawan Ibrahim  4 , Ifat Israel-Elgali  2 , Meitar Grad  2 , Rani Shlayem  2 , Ilan Youngster  2   4 , Mickey Scheinowitz  1   5 , Noam Shomron  2   3
Affiliations

Maximal and sub-maximal exercise tests alter PBMC microRNA expression: insights into sport- and sex-specific variations

Guy Shalmon et al. Front Physiol. .

Abstract

Introduction: Studying microRNAs in the immune cells of athletes offers a novel perspective on the molecular regulation of immune function and recovery, potentially uncovering strategies to enhance performance and resilience to physical stress. However, PBMC microRNA expression in endurance athletes, such as runners and cyclists, remains underexplored, especially with regard to sex differences.

Aims: (i) Assess sport- and sex-specific differences in PBMC microRNA expression induced by acute maximal and sub-maximal aerobic exercise in runners and cyclists and (ii) examine correlations between PBMC microRNAs and exercise performance. Methods: A total of 58 healthy athletes were included: 22 runners (9 females), 18 cyclists (9 females), and 18 active controls (9 females). Participants underwent VO2max and time-to-exhaustion tests, with blood samples collected pre- and post-exercise to analyze PBMC microRNA levels.

Results: Runners exhibited a higher microRNA response than cyclists or controls, with significant sex-based differences. After VO2max test, 279 microRNAs (255 upregulated) were altered in runners, compared to only seven microRNAs (none upregulated) in cyclists. Exercise intensity and duration had sport-specific effects on microRNA expression. Time-to-exhaustion in runners and weekly training volume in both groups were significantly associated with changes in PBMC microRNA profiles.

Conclusion: This study reveals that PBMC microRNA expression in response to maximal and sub-maximal exercise tests are sport- and sex-specific, providing new insights into the molecular adaptations of endurance athletes and their relationship to athletic performance.

Keywords: PBMC microRNA expression; cyclists; endurance athletes; females; peripheral blood mononuclear cells; runners.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

FIGURE 1
FIGURE 1
Volcano plot visualizing the differential expression of PBMC microRNAs (miRs) in response to Maximal (&/or Sub-Maximal exercise test(s) in runners compared to cyclists. The X-axis represents the adjusted log2Foldchange in microRNA expression between the two groups (runners vs. cyclists). A positive value of Log2FoldChange indicates an upregulation in microRNA expression (represented in red on the graph), while a negative value indicates a downregulation (represented in blue on the graph). The Y-axis represents the significance (−log10 [FDR]). The graph displays only microRNAs with statistical significance (FDR < 0.05).
FIGURE 2
FIGURE 2
Top 10 PBMC microRNAs (miRs) with the highest upregulation in expression induced by maximal and submaximal exercise in runners compared to cyclists. Log2FoldChange measures the change in microRNA expression, where the fold change between the expression levels of a microRNA in two groups (runners vs cyclists) is calculated. The ‘Log2’ transformation is applied to convert the change into logarithmic base two units, making the data more suitable for comparison. Figure generated with graphical assistance from AI software (ChatGPT, version 4); no AI involvement in data analysis or interpretation.
FIGURE 3
FIGURE 3
Volcano plot visualizing the association between time-to-exhaustion (min) and PBMC microRNA (miR) expression in runners. The results show differential expression of 195 PBMC microRNAs (FDR < 0.05; 75 upregulated) in relation to time-to-exhaustion. The X-axis represents the fold change in microRNA expression (Log2FoldChange), and the Y-axis represents the statistical significance (−log10 [FDR]). (A) The top 10 PBMC microRNAs with the highest upregulation in expression. (B) The top 10 PBMC microRNAs with the greatest downregulation in expression.
FIGURE 4
FIGURE 4
Volcano plot visualizing the association between weekly training volume (km) and PBMC microRNA (miR) expression in endurance athletes (runners and cyclists). The results show differential expression of 300 PBMC microRNAs (FDR < 0.05; 49 upregulated) in relation to weekly training volume. The X-axis represents the fold change in microRNA expression (Log2FoldChange), and the Y-axis represents the statistical significance (−log10 [FDR]). Results are valid for training distances of 50 km and above. (A) The top 10 PBMC microRNAs with the highest upregulation in expression. (B) The top 10 PBMC microRNAs with the greatest downregulation in expression.
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