目的 优化甘草次酸（GA）和叶酸（FA）共修饰双靶向pH敏感黄芩苷/姜黄素共载脂质体（GA/FA-pH-Lip@Bai/Cur）的处方工艺，并对其进行评价。方法 采用薄膜分散超声法制备GA/FA-pH-Lip@Bai/Cur，以包封率、不同pH释放介质中溶出曲线相似因子（f₂）、粒径和多分散指数（PDI）为评价指标，使用单因素考察和Box-Behnken设计-响应面法筛选出最佳处方工艺；对GA/FA-pH-Lip@Bai/Cur的外观形态、粒径、不同pH浓度的体外释放度等进行评价；通过溶血性实验评估GA/FA-pH-Lip@Bai/Cur的生物相容性；采用细胞CCK-8法考察黄芩苷、姜黄素、黄芩苷/姜黄素（质量比为5∶1）混合溶液、GA/FA-pH-Lip@Bai/Cur对肝癌细胞HepG2的体外增殖抑制作用；通过小鼠组织药物分布实验，考察GA/FA-pH-Lip@Bai/Cur体内的肝靶向性；探究GA/FA-pH-Lip@Bai/Cur （120、60、30 mg·kg^-1）对H22荷瘤小鼠的抑瘤率、HE染色后肿瘤组织病理学、血清肿瘤坏死因子-α（TNF-α）和白细胞介素-6（IL-6）水平的影响。结果 GA/FA-pH-Lip@Bai/Cur的最佳制备工艺为：黄芩苷与姜黄素质量比为5∶1，琥珀酸胆固醇单酯（CHEMS）与磷脂质量比为2∶10，DSPE-PEG 2000与磷脂质量比为4∶100，DSPE-PEG-GA与磷脂质量比为2∶100，水化体积为10 mL，磷脂与药物质量比7.95∶1，磷脂与胆固醇质量比6.53∶1，超声破碎时间为69 s。经响应面优化制得的GA/FA-pH-Lip@Bai/Cur黄芩苷包封率为90.90%，姜黄素包封率为90.97%，平均粒径82.5 nm，释放行为具有缓释特性。黄芩苷对HepG2细胞的半数抑制浓度（IC₅₀）大约是姜黄素的5倍，GA/FA-pH-Lip@Bai/Cur的IC₅₀较混合溶液低。体内药效评价结果表明，GA/FA-pH-Lip@Bai/Cur抑瘤率与5-氟脲嘧啶（5-FU）相当；给药组的肿瘤坏死细胞数量变少，肿瘤细胞的排列疏松，细胞呈现不同程度变小；血清中TNF-α和IL-6水平均较模型组显著降低（P＜0.01）。结论 制备的GA/FA-pH-Lip@Bai/Cur安全性好，肝靶向性高，具有良好的抑制肝肿瘤作用。

Objective To optimize the formulation process of dual-targeted pH-sensitive baicalin/curcumin co-loaded liposomes (GA/ FA-pH-Lip@Bai/Cur) and evaluate their properties. Methods GA/FA-pH-Lip@Bai/Cur was prepared using a thin film dispersion ultrasonication method. The evaluation criteria included encapsulation efficiency, similarity factor (f₂) of release curves in different pH media, particle size, and polydispersity index (PDI). The optimal formulation process was determined through single-factor testing and Box-Behnken design-response surface methodology to evaluate the liposomes' morphology, particle size, and in vitro release profiles at different pH levels. The biocompatibility of GA/FA-pH-Lip@Bai/Cur was assessed through a hemolysis test. The in vitro inhibitory effect of baicalin, curcumin, baicalin/curcumin (mass ratio 5∶ 1) mixed solution, GA/FA-pH-Lip@Bai/Cur on liver cancer cells (HepG2) was examined using the CCK-8 assay. The liver targeting of GA/FA-pH-Lip@Bai/Cur in vivo was investigated through a mouse tissue distribution experiment. Subsequently, pharmacological experiments were conducted to study the antitumor efficacy of GA/FA-pH-Lip@Bai/Cur on H22 tumor-bearing mice and its impact on histopathological changes of tumor tissue after HE staining, serum TNF-α and IL-6 levels. Results The optimal preparation method for GA/FA-pH-Lip@Bai/Cur was as follows: The ratio of baicalin to curcumin was 5∶1, the ratio of cholesteryl succinate (CHEMS) to phospholipids was 2∶10, the ratio of DSPE-PEG 2000 to phospholipids was 4∶ 100, the ratio of DSPE-PEG-GA to phospholipids was 2∶ 100, the hydration volume was 10 mL, the ratio of phospholipids to drugs was 7.95∶ 1, and the ratio of phospholipids to cholesterol was 6.53∶ 1, and the ultrasonic crushing time was 69 s. The optimized GA/FA-pH-Lip@Bai/Cur exhibited an encapsulation efficiency of 90.90% for baicalin and 90.97% for curcumin, with an average particle size of 82.5 nm and sustained-release behavior. The IC₅₀ of baicalin for HepG2 cells was approximately five times that of curcumin, and the IC₅₀ of GA/FA-pH-Lip@Bai/Cur was lower than that of the mixed solution. The in vivo pharmacodynamic evaluation results show that the tumor inhibition rate of GA/FA-pH-Lip@Bai/Cur was comparable to that of 5-fluorouracil (5-FU). The number of necrotic cells in the treated group decreased, the arrangement of tumor cells becomes loose, and the cells showed varying degrees of shrinkage, and the levels of TNF-α and IL-6 in the serum were significantly lower than those in the model group (P <0.01). Conclusion The GA/FA-pH-Lip@Bai/Cur prepared had good safety and high liver targeting, and has good inhibitory effects on liver tumors.

R943

2021年江苏省高校自然科学基金项目（21KJB360021）;2024年江苏高校青蓝工程优秀教学团队资助项目;2021年国家中医药管理局领军人才基金项目;泰州市科技支撑项目（TS202232、TS202325）;2023年江苏省大学生创新创业项目（202313981004Y、202313981011Y、202313981002Y、202313981003Y）