目的 响应面设计制备唾液酸（sialic acid，SA）修饰的绿原酸（chlorogenic acid，CA）脂质体（CA-SAL），考察其体外细胞毒性和摄取。方法 采用改良逆相乙醇注入法制备CA-SAL，以葡聚糖凝胶G-50柱离心法分离脂质体和游离药物，并通过HPLC法测定药物质量浓度，计算包封率。以包封率和载药量为考察指标，通过Box-Behnken响应面设计实验优化CA-SAL的处方和工艺，MTT法评价其对人肺癌A549细胞的细胞毒性，倒置荧光显微镜观察A549细胞对CA-SAL的摄取情况。结果 优化后的CA-SAL制备条件：氢化大豆磷脂与绿原酸的质量比为15∶1，水化温度60℃，超声功率400 W。CA-SAL平均粒径为（90.13±0.51）nm，多分散指数（PDI）为0.16±0.01，Zeta电位为（-25.3±0.5）mV；包封率为57.8%，RSD为0.1%。MTT实验结果显示，CA-SAL对A549细胞的增殖抑制作用显著强于绿原酸脂质体（CA-CL），细胞摄取实验表明A549细胞对唾液酸修饰脂质体的摄取更高。结论 通过响应面优化制备的CA-SAL粒径小、性质稳定。经唾液酸修饰后的脂质体可以增强人肺癌A549细胞的细胞摄取及体外细胞毒性。

Objective Sialic acid (SA)-modified chlorogenic acid (CA) liposomes (CA-SAL) was prepared by response surface design to investigate its in vitro cytotoxicity and uptake. Methods CA-SAL was prepared by a modified reverse-phase ethanol injection method. Sephadex G50 column was used to separate the CA-loaded liposomes and the free CA. The drug concentration was determined by HPLC method and the encapsulation efficiency was calculated. With encapsulation efficiency and drug loading as indicators, Box-Behnken response surface design experiments were used to optimize the prescription process of CA-SAL. The MTT method was used to evaluate the cytotoxicity of CA-SAL on human lung cancer cells A549. Inverted fluorescence microscope was used to investigate the uptake of CA-SAL by A549 cells. Results The optimized preparation conditions:hydrogenated soybean lecithin-CA ratio at 15:1, hydration temperature 60℃, ultrasonic power 400 W. The average particle size of CA-SAL was (90.13 ±0.51) nm, the polydispersity index (PDI) was 0.16 ±0.01, the zeta potential was (-25.3 ±0.5) mV, the encapsulation efficiency was 57.8%, RSD was 0.1%. MTT results showed that the inhibitory effect of CA-SAL on A549 cells was significantly greater than CA-CL. Greater cellular uptake of CA-SAL was observed compared with CA-CL. Conclusion CA-SAL prepared by response surface optimization has a uniform particle size and good stability. SA-modified CA-loaded liposomes could enhance cellular uptake and cytotoxicity of human lung cancer cell A549 in vitro.

R283.6

国家自然科学基金资助项目（81903808）；浙江省自然科学基金资助项目（LY21H280004）