data set and WB gel of "Reg3b in HIIT plasma promotes angiogenesis"

Background: Exercise therapy is the core intervention for managing peripheral arterial disease (PAD). Activating endogenous vascular regenerative mechanisms is crucial to overcoming the limitations of current therapeutic interventions. High-intensity interval training (HIIT) demonstrates superior proangiogenic effects compared to moderate-intensity continuous training (MICT); however, the underlying mechanisms remain poorly understood.Methods: Plasma samples from sedentary, MICT, and HIIT mice were intravenously administered to mice with hindlimb ischemia (HLI) to assess the effects on blood flow, muscle function, and capillary density. Proteomic profiling identified regenerating islet-derived protein 3 beta (Reg3b) as a candidate factor specific to HIIT. To evaluate macrophage M2 polarization, recombinant Reg3b (rReg3b) was applied to macrophages in vitro. Additionally, Reg3b-treated macrophages were intramuscularly transplanted into ischemic limbs. AAV-mediated pancreas-specific Reg3b overexpression (oeReg3b) was performed to investigate its effects on macrophage polarization and vascular recovery. The role of EXTL3/mTORC2/CPT1a-dependent lipid metabolism in regulating Reg3b-induced macrophage polarization was examined using siRNA and specific inhibitors.Results: Plasma from exercised mice, particularly those undergoing HIIT, significantly enhanced perfusion recovery, angiogenesis, and muscle function compared to sedentary controls. Notably, Reg3b levels in HIIT-enriched plasma positively correlated with exercise capacity and facilitated the polarization of CD206⁺ M2 macrophages. Macrophages treated with Reg3b or mice overexpressing oeReg3b displayed enhanced post-ischemic angiogenesis and functional recovery. Mechanistically, Reg3b interacted with EXTL3 and upregulated Rictor (a component of mTORC2), PPARγ, and CPT1a in macrophages. Inhibition of EXTL3, mTORC2, or CPT1a abolished Reg3b-induced M2 polarization and VEGF secretion.Conclusions: Exercise-derived plasma exhibits transferable proangiogenic benefits, with HIIT producing the most potent effects. Pancreatic-derived Reg3b emerges as a pivotal circulating factor induced by HIIT, promoting macrophage M2 polarization and vascular regeneration via mTORC2-mediated fatty acid metabolism.