Fresh blood was collected from the hearts of mice in each group. The serum was collected via centrifugation at 10,000 × g at 4°C for 15 min. The secretion of serum inflammatory cytokines (IL-1β, IL-6, TNF-α, and M-CSF) was measured using ELISA MAX Deluxe (BioLegend, San Diego, CA, United States) and Quantikine ELISA Kit (R&D systems, Minneapolis, MN, United States), following the manufacturer’s instructions. The absorbance at 450 nm was measured using a microplate reader (SH-1000 Lab; Corona Electric Co., Hitachinaka, Japan). All tests were performed in two wells per individual. The serum concentrations of IL-1β, IL-6, TNF-α, and M-CSF were quantified, and the concentrations were set to 0 if the results were below the detection limit.

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