Exercise and Probiotic-Induced and Exosomal Modulations:A 12-Week Intervention Study in Healthy Individual

Background: This study investigated the combined effects of a 12-week training program and probiotic supplementation on epigenetic modifications and exosomal Hsp60 expression. Physical activity (PA) is known to influence gene regulation through mechanisms such as DNA methylation and non-coding RNA modulation, as well as through intercellular communication via exosomes. Nutritional interventions, particularly probiotics like Lactobacillus fermentum, may also modulate these pathways, potentially enhancing Hsp60 expression. The study aimed to explore these interactions in both sedentary individuals and trained athletes. Methods: Twelve adults were divided into two groups: six physically active individuals and six sedentary controls. Blood samples were collected before and after an acute exercise session at baseline (T0) and after twelve weeks of daily probiotic supplementation. Epigenetic analysis was performed using MeSAP-PCR and MSRE-PCR to assess global and gene-specific DNA methylation. Plasma exosomes were isolated and analyzed for Hsp60 expression via Western blot, and cytokine levels were quantified using MagPix® technology. Results: Significant differences in epigenetic profiles emerged between active and sedentary participants. Regular physical activity was associated with favorable global hypermethylation and selective promoter hypomethylation in genes involved in metabolism, mitochondrial function, and anti-inflammatory responses. Sedentary individuals showed the opposite pattern. Additionally, exercise and probiotics modulated Hsp60 levels in plasma exosomes, suggesting a synergistic role in enhancing intercellular communication, possibly via muscle-derived exosomes. Conclusion: Exercise and probiotic supplementation act as synergistic, non-pharmacological modulators of epigenetic and exosomal pathways. These findings support their potential in personalized health strategies aimed at improving metabolic function and preventing chronic disease.