Sonodynamic Therapy Attenuates TLR4/NF-κB Signaling in Epicardial Adipose Tissue of Atrial Fibrillation: Insights from Animal Models, scRNA-seq, and Mendelian Randomization,

Atrial fibrillation (AF) is the most common arrhythmia leading to cardioembolic stroke, with inflammation of epicardial adipose tissue (EAT) contributing to its pathogenesis. However, current therapies cannot specifically target EAT inflammation to alleviate AF. This study investigated the effects of sonodynamic therapy (SDT) on reducing macrophage-mediated inflammation in the EAT of rabbit AF models and explored its potential clinical application in AF patients. Rabbits were assigned to sham, paced (600 bpm for 4 w), and SDT groups. Macrophage infiltration and collagen deposition were evaluated using RAM11 immunohistochemistry and Masson staining, respectively. Transcriptomic analysis and network pharmacology identified TLR4/NF-κB pathway genes (including CXCL8 and CCL2) as SDT targets for AF treatment, which were validated by Western blot and RT-qPCR. Single-cell sequencing of human EAT and right atrial appendage confirmed the expression of these SDT-targeted genes in macrophages from AF patients. Mendelian randomization demonstrated that elevated plasma CXCL8, but not CCL2, was associated with an increased risk of atrial fibrillation. SDT reduced macrophage infiltration, fibrosis, and TLR4/NF-κB-mediated inflammation, supporting its therapeutic potential in AF via modulation of EAT inflammation.