Emerging evidence indicates that macrophage functional polarization is critically involved in the development of atherosclerosis (AS). Here, we examined the role of 5-aminolaevulinic acid (ALA)-mediated non-lethal sonodynamic therapy (NL-SDT) in macrophage-subset polarization and atherosclerotic lesion stability and explored the potential underlying mechanisms. Using Western diet-fed apolipoprotein E (apoE)^−/− and green fluorescent protein (GFP)-positive bone marrow (BM) chimeric mouse models, we demonstrated that NL-SDT promoted phenotypic switching of both BM-derived and resident macrophages from M1 to M2 and significantly inhibited AS progression. Further mechanistic studies indicated that NL-SDT enhanced macrophage differentiation toward the M2 phenotype by activating the reactive oxygen species (ROS)–5′ AMP-activated protein kinase (AMPK)–mammalian target of rapamycin complex 1 (mTORC1)–autophagy signaling pathway in murine BM-derived M1 macrophages (BMDM1s). Moreover, NL-SDT drastically reduced lipid droplets, mainly by promoting apoAI-mediated cholesterol efflux in vitro. Specifically, administration of pharmacological inhibitors to the animal model showed a reciprocal effect on NL-SDT-induced macrophage polarization. These findings indicate that NL-SDT engages a virtuous cycle that enhances M1-to-M2 polarization, cholesterol efflux, and anti-inflammatory reactions in advanced plaque in vivo and in BMDM1s in vitro by activating the ROS–AMPK–mTORC1–autophagy pathway. This discovery might help elucidate the mechanism underlying NL-SDT as a potential treatment to prevent atherothrombotic events.

新兴证据表明，巨噬细胞功能极化与动脉粥样硬化（AS）的发展至关重要。在这里，我们检查了5-氨基松香酸（ALA）介导的非致死声波动力疗法（NL-SDT）在巨噬细胞亚群极化和动脉粥样硬化病变稳定性中的作用，并探讨了潜在的潜在机制。使用西方饮食喂养的载脂蛋白E（apoE）^-/-和绿色荧光蛋白（GFP）阳性骨髓（BM）嵌合小鼠模型，我们证明了NL-SDT促进了BM来源的和驻留的巨噬细胞从M1到M2的表型转换，并显着抑制了AS的进展。进一步的机理研究表明，NL-SDT通过激活活性氧（ROS）-5'AMP激活的蛋白激酶（AMPK）-雷帕霉素复合物1（mTORC1）-自噬信号转导途径，增强了巨噬细胞向M2表型的分化。鼠BM衍生的M1巨噬细胞（BMDM1s）。另外，NL-SDT急剧降低脂滴，主要通过促进的apoAⅠ介导的胆固醇流出体外。具体而言，将药理抑制剂施用于动物模型显示出对NL-SDT诱导的巨噬细胞极化的相互影响。这些结果表明，NL-SDT接合良性循环增强M1至M2极化，胆固醇流出，并且在高级斑块抗炎反应在体内和在BMDM1s体外通过激活ROS-AMPK-的mTORC1的自噬途径。这一发现可能有助于阐明NL-SDT的潜在机制，作为预防动脉粥样硬化血栓形成事件的潜在治疗方法。