Table_2_Physical Activity as a Preventive Lifestyle Intervention Acts Through Specific Exosomal miRNA Species—Evidence From Human Short- and Long-Term Pilot Studies.DOCX

Exercise initiates systemic adaptation to promote health and prevent various lifestyle-related chronic diseases. Emerging evidence suggests that circulating exosomes mediate some of the beneficial effects of exercise via the transfer of microRNAs between tissues. Yet to date, a comprehensive profile of the exosomal miRNA (exomiR) content released following short-term (0.5 year in this study) and long-term (25 + years in this study) regular bouts of exercise is still lacking. However, a better understanding of these miRNA species would assist in clarifying the role of regular exercise at the molecular level in the prevention of chronic diseases. In the present pilot studies we analyzed serum exomiR expression in healthy young, sedentary participants (n = 14; age: 23 ± 2 years) at baseline and following a half year-long moderate-intensity regular exercise training. We also analyzed serum exomiR expression in older, healthy trained participants (seniors, n = 11; age: 62 ± 6 years) who engaged in endurance activities for at least 25 years. Following the isolation and enrichment of serum exosomes using Total Exosome Isolation Reagent (TEI) their exomiR levels were determined using the amplification-free Nanostring platform. Hierarchical cluster analysis revealed that the majority of exomiRs overlap for short-term (0.5 year in this study) and long-term (25 + years in this study) regular bouts of exercise. The top 12 significantly altered exomiRs (let-7a-5p; let-7g-5p; miR-130a-3p; miR-142-3p; miR-150-5p; miR-15a-5p; miR-15b-5p; miR-199a-3p; miR-199b-3p; miR-223-3p; miR-23a-3p, and miR-451a-3p) were used for further evaluation. According to KEGG pathway analysis a large portion of the exomiRs target chronic diseases including cancer, neurodegenerative and metabolic diseases, and viral infections. Our results provide evidence that exosomal miRNA modulation is the molecular mechanism through which regular exercise prevents various chronic diseases. The possibility of using such exomiRs to target diseases is of great interest. While further validation is needed, our comprehensive exomiR study presents, for the first time, the disease-preventive molecular pattern of both short and long-term regular exercise.

运动可引发机体系统性适应，从而促进健康并预防多种生活方式相关慢性疾病。越来越多的研究证据表明，循环外泌体（exosomes）可通过组织间微小RNA（microRNAs, miRNAs）的传递，介导运动带来的部分益处。然而截至目前，学界仍缺乏针对短期（本研究中为0.5年）与长期（本研究中为25年以上）规律运动后释放的外泌体微小RNA（exosomal miRNA, exomiR）含量的全面分析图谱。但深入解析这类miRNA分子，将有助于阐明规律运动在慢性疾病预防中的分子层面作用机制。在本项预实验中，我们分析了健康青年久坐受试者（n=14；年龄：23±2岁）在基线状态以及为期半年的中等强度规律运动训练后的血清exomiR表达水平；同时还分析了至少坚持耐力运动达25年以上的健康老年受训受试者（n=11；年龄：62±6岁）的血清exomiR表达水平。本研究采用总外泌体分离试剂（Total Exosome Isolation Reagent, TEI）对血清外泌体进行分离与富集后，通过无扩增纳米串（Nanostring）检测平台测定其exomiR表达水平。层次聚类分析结果显示，短期（0.5年）与长期（25年以上）规律运动后，多数外泌体微小RNA的表达谱存在重叠。本研究选取12个表达变化最显著的exomiR（let-7a-5p、let-7g-5p、miR-130a-3p、miR-142-3p、miR-150-5p、miR-15a-5p、miR-15b-5p、miR-199a-3p、miR-199b-3p、miR-223-3p、miR-23a-3p及miR-451a-3p）开展后续验证分析。经KEGG通路分析发现，大部分上述exomiR的靶基因涉及癌症、神经退行性疾病、代谢性疾病及病毒感染等慢性疾病领域。本研究结果证实，外泌体微小RNA的表达调控是规律运动预防多种慢性疾病的核心分子机制。利用这类exomiR进行疾病靶向干预的可能性也引发了学界广泛关注。尽管仍需开展进一步验证，但本项全面的exomiR研究首次揭示了短期与长期规律运动的疾病预防分子模式。