Exercise therapy exerts pleiotropic benefits in disease prevention and treatment, yet the underlying molecular mechanisms remain unclear. Exerkines, signaling molecules released by tissues like skeletal muscle, cardiac muscle, adipose, and liver during exercise, mediate inter-organ crosstalk and tissue adaptation. These exerkines, including peptides, proteins, metabolites, lipids, and nucleic acids, play crucial roles in health and disease. Understanding their molecular mechanisms is essential for developing targeted exercise programs and therapeutic strategies. Exerkines are secreted in response to acute or chronic exercise, with some being mobilized during a single session and others requiring repeated exercise. Their secretion is regulated by various molecular triggers, including calcium-dependent pathways, hypoxia, and shear stress on endothelial cells. Exerkines can be secreted via conventional pathways or through extracellular vesicles (EVs), which facilitate long-distance signaling and tissue crosstalk. Exerkines exhibit diverse biological functions, including metabolic regulation, immune modulation, and tissue remodeling. For example, lactate, a metabolite secreted during exercise, interacts with other biomolecules to influence feeding behavior and obesity. Succinate, another exerkine, is mobilized in response to exercise and contributes to tissue adaptation. Non-coding RNAs, such as miRNAs and lncRNAs, also play significant roles in exerkine signaling, influencing gene expression and cellular processes. Exerkine signaling is mediated through receptor-dependent and receptor-independent mechanisms. Receptor-dependent pathways involve interactions with specific receptors, leading to intracellular signaling cascades that regulate gene expression and cellular adaptation. Receptor-independent mechanisms include direct delivery of exerkines to target cells via EVs or passive diffusion across cell membranes. The clinical implications of exerkine signaling are significant, with potential applications in disease prevention, targeted exercise therapy, and the development of exercise mimetics. Understanding exerkine kinetics and dynamics is crucial for translating molecular insights into therapeutic strategies. Exerkine research provides a foundation for improving public health through targeted exercise programs and novel pharmaceutical approaches.Exercise therapy exerts pleiotropic benefits in disease prevention and treatment, yet the underlying molecular mechanisms remain unclear. Exerkines, signaling molecules released by tissues like skeletal muscle, cardiac muscle, adipose, and liver during exercise, mediate inter-organ crosstalk and tissue adaptation. These exerkines, including peptides, proteins, metabolites, lipids, and nucleic acids, play crucial roles in health and disease. Understanding their molecular mechanisms is essential for developing targeted exercise programs and therapeutic strategies. Exerkines are secreted in response to acute or chronic exercise, with some being mobilized during a single session and others requiring repeated exercise. Their secretion is regulated by various molecular triggers, including calcium-dependent pathways, hypoxia, and shear stress on endothelial cells. Exerkines can be secreted via conventional pathways or through extracellular vesicles (EVs), which facilitate long-distance signaling and tissue crosstalk. Exerkines exhibit diverse biological functions, including metabolic regulation, immune modulation, and tissue remodeling. For example, lactate, a metabolite secreted during exercise, interacts with other biomolecules to influence feeding behavior and obesity. Succinate, another exerkine, is mobilized in response to exercise and contributes to tissue adaptation. Non-coding RNAs, such as miRNAs and lncRNAs, also play significant roles in exerkine signaling, influencing gene expression and cellular processes. Exerkine signaling is mediated through receptor-dependent and receptor-independent mechanisms. Receptor-dependent pathways involve interactions with specific receptors, leading to intracellular signaling cascades that regulate gene expression and cellular adaptation. Receptor-independent mechanisms include direct delivery of exerkines to target cells via EVs or passive diffusion across cell membranes. The clinical implications of exerkine signaling are significant, with potential applications in disease prevention, targeted exercise therapy, and the development of exercise mimetics. Understanding exerkine kinetics and dynamics is crucial for translating molecular insights into therapeutic strategies. Exerkine research provides a foundation for improving public health through targeted exercise programs and novel pharmaceutical approaches.