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目的:制备载miR-33反义核苷酸(ANM33)的双靶标脂质超声微泡(HA-PANBs)，体外评价其分阶段靶向泡沫细胞并实现细胞内精准释药的可行性。方法:设计以纳米泡NBs为微泡核心，透明质酸(hyaluronic acid，HA)第一阶段靶向受损内皮细胞，适配子PM1第二阶段靶向泡沫细胞，ANM33为治疗因子的双靶标脂质超声微泡。完成脂质泡表征的同时检测其稳定性及体外造影能力；随后建立受损人脐静脉内皮细胞(HUVEC)和巨噬细胞(RAW264.7，MΦ)共培养模型，结合流式细胞术、油红O染色和小动物活体成像仪等评价HA-PANBs体外精准定位泡沫细胞并释放ANM33的能力。结果:制备的双靶标脂质超声微泡HA-PANBs形态规则，稳定性好，粒径为(1 357.53±140.20)nm，表面电位为(－5.61±0.73)mV，HA、PM1及ANM33均有效连接。在受损HUVEC/MΦ共培养模型中，HA-PANBs靶向下层泡沫细胞的效果最佳，且有效摄取量高达(80.65±2.12)%，较NBs组高56.74%；油红O染色显示HA-PANBs组泡沫细胞的胆固醇外排能力明显优于NBs组，差异有统计学意义[(629.80±21.99)a.u.比(1 071.00±55.49)a.u.， P<0.05]。 结论:构建的双靶标脂质超声微泡HA-PANBs能精准靶向泡沫细胞并显著增强其胆固醇外排能力，为实现动脉粥样硬化的早期无创诊疗提供了新策略。

Objective:To prepare dual-targeted lipid ultrasound microbubbles loaded with ANM33 (HA-PANBs) and evaluate its feasibility in targeting foam cells by stages and achieving precise intracellular drug release in vitro. Methods:The dual-targeteded lipid ultrasound microbubbles were designed with nanobubbles (NBs) as the microbubble core, hyaluronic acid (HA) as the first-stage targeting ligand for damaged endothelial cells, aptamer PM1 as the second-stage targeting moiety for foam cells, and ANM33 as the therapeutic factor. Simultaneously with the characterization of the lipid bubbles, the stability and in vitro contrast-enhanced ultrasound imaging capability were detected. Then a co-culture model of damaged human umbilical vein endothelial cells (HUVEC) and macrophages (RAW264.7, MΦ) was established, combined with flow cytometry, oil red O staining and small animal in vivo imaging to evaluate the ability of HA-PANBs in targeting foam cells precisely and releasing ANM33. Results:The HA-PANBs exhibited regular morphology and good structural stability, with a particle size of (1 357.53±140.20)nm and a surface potential of (-5.61±0.73)mV. HA, PM1 and ANM33 were effectively connected. In the damaged HUVEC/MΦ co-culture system, the HA-PANBs group demonstrated the best targeting effect on foam cells, with an effective uptake of (80.65±2.12)%, which was 56.74% higher than that of the NBs group. Oil red O staining revealed that the cholesterol efflux capacity of foam cells in the HA-PANBs group was significantly better than that in the NBs group, the results were statistically different [(629.80±21.99) a.u.vs (1 071.00±55.49)a.u., P<0.05]. Conclusions:The dual-targeted lipid ultrasound microbubbles (HA-PANBs) can precisely target foam cells and significantly enhance their cholesterol efflux, providing a new strategy for the early non-invasive diagnosis and treatment of atherosclerosis.