藤黄酸-重组高密度脂蛋白纳米粒的制备及评价

Preparation and evaluation of gambogic acid-loaded reconstituted high density lipoprotein nanoparticles

采用胆酸钠法制备藤黄酸(GA)重组高密度脂蛋白纳米粒(GA-rHDL-NPs),并测定其形态、粒径、Zeta电位、包封率和载药量等理化性质,同时以透析法研究制剂的体外释药特性,细胞实验考察肿瘤细胞对其的摄取能力,溶血性试验和家兔耳缘静脉刺激性试验评价其静脉注射的安全性。结果显示,GA-rHDL-NPs外观呈类球形,平均粒径为(113.53±2.50)nm,Zeta电位为-(29.48±0.05)mV,包封率和载药量分别为(95.30±0.37)%和(8.51±0.95)%,其水分散液4℃放置一个月稳定;GA-rHDL-NPs体外24 h和72 h的累积释放量分别为24.3%和73.6%,其肝肿瘤细胞(HepG2)摄取能力明显强于正常肝细胞(L02),且无明显溶血作用和耳缘静脉刺激性反应。研究结果表明,GA-rHDL-NPs的药物包封率高,性质稳定,药物释放缓慢,分散性和安全性良好,可供静脉注射使用,并具有良好的肿瘤细胞摄取能力。

Gambogic acid（GA）-loaded reconstituted high-density lipoprotein nanoparticles（GA-rHDL-NPs） were prepared by sodium cholate method.The physicochemical properties such as morphology,particle size,Zeta potential,entrapment efficiency and drug-loading capability were evaluated.Dialysis method was employed to investigate in vitro release of GA-rHDL-NPs.In addition,the cellular uptake experiments were performed to evaluate the tumor cellular targeting ability of GA-rHDL-NPs,and safety evaluation following the intravenous injection of GA-rHDL-NPs was carried out using hemolysis testing and irritation testing upon rabbit ear vein.It was found that the well-formulated GA-rHDL-NPs displayed regular spherical shape with particle size of（113.53±2.50） nm,Zeta potential of-（29.48±0.05） mV,entrapment efficiency of（95.30±0.37）% and drug-loading of（8.51±0.95）%.The GA-rHDL-NPs solution was found to be stable after one-month storage at 4 °C.In vitro 24-h and 72-h accumulative releases of GA from GA-rHDL-NPs were approximately 24.3% and 73.6%,respectively.It revealed that there was much more uptake of GA-rHDL-NPs by HepG2 cells than by L02 cells.GA-rHDL-NPs also exhibited less local venous irritation and hemolysis than did the unencapsulated drug group.Therefore,the GA-rHDL-NPs possessed accepted entrapment efficiency,stability,sustained-release,dispersion and safety properties,and tumor cell uptake,which might be suitable for iv injection and cancer therapy.