目的 通过处方优化构建稳定性高、包封率优的灵芝酸A脂质体（ganoderic acid A liposomes，GA-Lips），并评价其细胞摄取效率及体外抗肿瘤效应。方法 采用薄膜分散法制备GA-Lips，通过调控药物、卵磷脂和胆固醇比例进行处方优化，并开展粒径、ζ电位及透射电子显微镜（transmission electron microscope，TEM）进行形貌表征。利用超滤离心法考察GA-Lips体外释放，结合介质稳定性、稀释稳定性和长期稳定性评估GA-Lips制剂稳定性。通过溶血实验评价血液相容性；采用共聚焦显微镜与流式细胞术分析细胞摄取行为；以CCK-8法测定体外抗增殖作用，并采用Annexin V-FITC/PI流式细胞术检测细胞凋亡。结果 优化后的GA-Lips粒径为（74.52±1.00）nm、PDI为0.26±0.01、ζ电位为（-46.75±1.61）mV，形貌均一。制剂呈缓释特性，在多种介质环境与稀释梯度下均保持良好稳定性，无显著溶血。GA-Lips显著提高HepG2肿瘤细胞的摄取水平，共聚焦显微镜及流式细胞术均得到验证。CCK-8结果显示，GA-Lips对HepG2细胞有显著的细胞毒性；流式分析表明，GA-Lips明显提高HepG2细胞凋亡比例。结论 处方优化构建的GA-Lips具有良好稳定性和安全性，可显著提升灵芝酸A的细胞摄取与体外抗肿瘤活性，为其进一步机制研究及纳米递药系统开发提供实验依据。

Objective To develop formulation-optimized ganoderic acid A liposomes (GA-Lips) with improved stability and high encapsulation efficiency, and to evaluate their cellular uptake efficiency and in vitro antitumor activity. Methods GA-Lips were prepared using the thin-film hydration method, and formulation parameters were optimized by varying the ratios of GAA, phospholipids, and cholesterol. Particle size, ζ potential, and morphology were characterized. In vitro drug release and formulation stability were evaluated under different media, dilution, and storage conditions. Hemocompatibility was assessed by hemolysis assays. Cellular uptake efficiency was examined using confocal laser scanning microscopy and flow cytometry. Antiproliferative effects and apoptosis induction in HepG2 cells were analyzed using CCK-8 and Annexin V-FITC/PI assays, respectively. Results The optimized GA-Lips exhibited a uniform spherical morphology with a mean particle size of (74.52 ± 1.00) nm, a PDI of 0.26 ± 0.01, and a ζ potential of (-46.75 ± 1.61) mV. The formulation showed sustained drug release, excellent stability under various test conditions, and negligible hemolytic activity. GA-Lips significantly enhanced cellular uptake efficiency in HepG2 cells compared with free GAA. Moreover, GA-Lips exhibited pronounced cytotoxic effects and markedly increased apoptosis induction. Conclusion The optimized GA-Lips demonstrated favorable stability and hemocompatibility, effectively enhancing the cellular uptake efficiency and in vitro antitumor efficacy of GAA. These results support the potential of GA-Lips as a promising nanocarrier system for ganoderic acid A and provide a foundation for further in vivo and mechanistic studies.

R283.6

中国科协青年托举工程人才项目（YESS20240482）