Journal: Journal of Sport and Health Science

Article Title: Long-term aerobic exercise enhances circulating exosomal miR-214-3p to promote endothelial progenitor cell-mediated repair of endothelial damage induced by obesity

doi: 10.1016/j.jshs.2025.101094

Figure Lengend Snippet: Circulating exosomal miR-214-3p regulated the function of EPC in both humans and rats with obesity. (A and B) Targeted transcriptome sequencing of miRNAs within circulating exosomes derived from (A) humans and (B) rats unveiled the regulatory impact of aerobic exercise on miRNA expression profiles in these vesicles. The left panel shows a volcano plot of differentially expressed genes, and the right panel shows a heatmap representation of the transcriptome sequencing data, illustrating the expression patterns of differentially expressed genes in circulating exosomes. (C and D) qPCR was used to validate the relative expression levels of miR-214-3p in circulating exosomes derived from (C) humans ( n = 3 for each group; * p < 0.05, Exercise vs . Control) and (D) rats ( n : 6–12 for each group; * p < 0.05, HC vs . NC; # p < 0.05, HE vs . HC). (E) qPCR validation of miR-214-3p expression in miR-214-3p-overexpressing EPC ( n = 3 for each group). ** p < 0.01, miR-214-3p mimics vs . mimics NC; ## p < 0.01, miR-214-3p mimics vs . Control. (F) Summary data for cell vitality level among groups ( n : 10–12 for each group). *** p < 0.001, miR-214-3p mimics vs . mimics NC. (G and H) Summary data (G) and representative images (H) of wound healing in the scratch assay, showcasing the migration level of EPC among groups ( n = 4 for each group). * p < 0.05, miR-214-3p mimics vs . mimics NC. EPC = endothelial progenitor cells; HC = the high-fat diet with sedentary group; HE = the high-fat diet with exercise group; mimics NC = mimics negative control; miR = microRNA; NC = the normal diet with sedentary group; qPCR = quantitative polymerase chain reaction.

Article Snippet: Total RNAs from tissues, cells, and exosomes were extracted using Trizol (R0016; Beyotime Biotech, Shanghai, China), following the manufacturer’s recommendations. mRNA samples underwent reverse transcription using the Evo M-MLV Kit (AG11705; Accurate Biotechnology, Changsha, China), while miRNA samples were reversely transcribed using the All-in-OneTM miRNA qPCR Kit (QP115; iGene Biotechnology, Guangzhou, China), following the provided instructions.

Techniques: Sequencing, Derivative Assay, Expressing, Control, Biomarker Discovery, Wound Healing Assay, Migration, Negative Control, Real-time Polymerase Chain Reaction

Journal: Journal of Sport and Health Science

Article Title: Long-term aerobic exercise enhances circulating exosomal miR-214-3p to promote endothelial progenitor cell-mediated repair of endothelial damage induced by obesity

doi: 10.1016/j.jshs.2025.101094

Figure Lengend Snippet: miR-214-3p plays an essential role in exercise-mediated protection against obesity-induced EPC dysfunction in vivo . (A) Dynamic weight change curves of rats. After randomization at Week 10, rats underwent treadmill exercise until the end of Week 18 ( n : 5–7 for each group). * p < 0.05, KO + HE vs . WT + NC; ### p < 0.001, WT + HC vs . WT + NC; ††† p < 0.001, WT + HE vs . WT + NC; ‡‡‡ p < 0.001, WT + HE vs . WT + HC; §§ p < 0.01, KO + HE vs . WT + HE. (B) Summary data for Lee's index among groups at Week 10 and Week 18 ( n : 5–7 for each group). Left panel: ** p < 0.01, KO + HE vs . WT + NC; ### p < 0.001, WT + HC vs . WT + NC; ††† p < 0.001, WT + HE vs . WT + NC; Right panel: *** p < 0.001, KO + HE vs . WT + HC; ### p < 0.001, WT + HC vs . WT + NC; ‡‡‡ p < 0.001, WT + HE vs . WT + HC. (C) Summary data for acetylcholine (ACh)-induced, endothelium-dependent relaxation in mesenteric arteries among groups ( n : 5–6 for each group). * p < 0.05, WT + HE vs . KO + HE; # p < 0.05, ## p < 0.05, WT + HE vs . WT + HC. (D) Summary data for EC 50 values in mesenteric arteries among groups in response to ACh ( n : 5–6 for each group). * p < 0.05, KO + HE vs . WT + HE; ## p < 0.01, WT + HE vs . WT + HC; ‡‡ p < 0.01, WT + HC vs . WT + NC. (E) Cell proliferation assays demonstrated that exosomes derived from the WT + HC group exhibited a diminished capacity to promote EPC proliferation compared to the WT + NC group in rats. In contrast, exosomes derived from the WT + HE group significantly enhanced EPC proliferation, which was abolished by knocking out miR-214-3p in rats ( n = 10 for each group). *** p < 0.001, KO + HE vs . WT + HE; ### p < 0.001, WT + HE vs . WT + HC; ‡‡ p < 0.01, WT + HC vs . WT + NC. (F) Scratch assay results showed that exosomes derived from the WT + HC group exhibited a diminished capacity to enhance EPC migration rates compared to the WT + NC group in rats. In contrast, exosomes derived from the WT + HE group significantly enhanced EPC migration rates, which was abolished by knocking out miR-214-3p in rats ( n = 6 for each group). *** p < 0.001, KO + HE vs . WT + HE; ### p < 0.001, WT + HE vs . WT + HC; ‡‡ p < 0.01, WT + HC vs . WT + NC. (G) Representative images of wound healing in the scratch assay, showcasing the migratory response of rat EPC. (H) qPCR analyses of pre-miR-214-3p and miR-214-3p in tissues from obese rats with and without exercise training ( n : 4–6 for each group). *** p < 0.001, HE vs . HC. EC 50 = half maximal effective concentration; EPC = endothelial progenitor cells; HC = the high-fat diet with sedentary group; HE = the high-fat diet with exercise group; KO + HE = the knockout + high-fat diet with exercise group; miR = microRNA; pre-miR = precursor microRNA; WT + HC = the wild-type + high-fat diet with sedentary group; WT + HE = the wild-type + high-fat diet with exercise group; WT + NC = the wild-type + normal diet with sedentary group.

Article Snippet: Total RNAs from tissues, cells, and exosomes were extracted using Trizol (R0016; Beyotime Biotech, Shanghai, China), following the manufacturer’s recommendations. mRNA samples underwent reverse transcription using the Evo M-MLV Kit (AG11705; Accurate Biotechnology, Changsha, China), while miRNA samples were reversely transcribed using the All-in-OneTM miRNA qPCR Kit (QP115; iGene Biotechnology, Guangzhou, China), following the provided instructions.

Techniques: In Vivo, Derivative Assay, Wound Healing Assay, Migration, Concentration Assay, Knock-Out