目的 利用脂质体可以共载水溶性物质和油溶性物质的特点，制备共载HBx-siRNA与五味子油阳离子脂质体纳米粒，并进行制备工艺优化以及体外抑制乙型肝炎的药效学研究。方法 采用单因素考察和星点设计-效应面法，优化共载HBx-siRNA与五味子油阳离子脂质体纳米粒的制备工艺，采用电位粒径仪测定其粒径和电位，透射电子显微镜（TEM）观察其形态。通过MTT比色法、细胞划痕实验、细胞摄取实验、ELISA法检测乙肝核心抗原（HBeAg）和乙肝表面抗原（HBsAg）表达水平等实验进行体外药效学研究。结果 确定最佳工艺为（2，3-二油氧基-丙基）三甲基氯乙铵（DOTAP）与胆固醇的质量比为3∶1，五味子油与脂质体的质量比为1∶5，旋蒸温度30℃，超声时间70 min，包封率可达84.08%，测得粒径为（134.000±0.035）nm，Zeta电位为（44.000±0.027）mV，形状呈不规则的圆形。体外药效学实验表明，共载HBx-siRNA与五味子油阳离子脂质体纳米粒体外抑制乙型肝炎的效果显著。结论 成功制备了新型脂质体纳米给药系统。体外药效学实验表明五味子油具有较好的协同HBx-siRNA抑制乙型肝炎的作用。

Objective To make use of the characteristic that liposome can co-carry water-soluble substance and oil-soluble substance, the co-carried HBx-siRNA and Wuweizi (Schisandra Chinensis Fructus, SCF) oil cationic liposome nanoparticles were prepared, and the preparation process is optimized as well as the in vitro pharmacodynamic study on the inhibition of hepatitis B. Methods The single factor test and central composite design-response surface methodology were used to optimize the preparation process of cationic liposome nanoparticles co-loaded with HBx-siRNA and SCF oil. The particle size and potential were measured by a potential particle size analyzer, and the morphology was observed by a transmission electron microscope (TCM). In vitro pharmacodynamic studies were conducted through experiments such as MTT colorimetric method, cell scratch test, cell uptake test, and ELISA method for detecting the expression levels of hepatitis b core antigen (HBeAg) and hepatitis b surface antigen (HBsAg). Results The optimal conditions were determined as follows: The mass ratio of (2,3-dioleoyloxy-propyl)-trimethylam monium (DOTAP) to cholesterol was 3∶1, the mass ratio of SCF oil to liposome was 1∶5, steaming temperature was 30 ℃, and ultrasonic time was 70 min. The encapsulation efficiency reached 84.08%, and the measured particle size was (134.000 ±0.035) nm, Zeta potential was (44.000 ±0.027) mV, and the shape was irregular and round. In vitro pharmacodynamics experiments showed that HBX-siRNA co-loaded with SCF oil group had the best efficacy. The co-carrier of HBx-siRNA and SCF oil cationic liposome nanoparticles had significant inhibitory effect on hepatitis B in vitro. Conclusion A new liposome nanometer drug delivery system was successfully prepared. In vitro pharmacodynamic experiments showed that SCF oil had a good synergistic effect with HBx-siRNA for the inhibition of hepatitis B.

R283.6

国家自然科学基金资助项目（81603418）