甘露糖修饰姜黄素/人参皂苷Rb1脂质体的处方优选及体外脑靶向性评价

Formulation Optimization and in vitro Brain Targeting Evaluation of Mannose-modified Curcumin/ginsenoside Rb1 Liposomes

目的:优化甘露糖(MAN)修饰姜黄素(Cur)/人参皂苷Rb1(GS-Rb1)共载脂质体(Lip)的处方,考察其体外靶向能力。方法:通过薄膜分散法-硫酸铵梯度法制备MAN-Cur/GS-Rb1-Lips,以包封率为考察指标,采用星点设计响应面法确定MAN-Cur/GS-Rb1-Lips最佳处方。采用荧光显微镜法考察了MAN-Cur/GS-Rb1-Lips在小鼠脑微血管内皮细胞(bEND.3)和小鼠来源神经瘤母细胞(N2a)主动靶向性。结果:通过Box-Behnken响应面法,最终确定脂质体的最优处方为DSPE-PEG_(2000)-MAN 2 mg、卵磷脂88 mg、胆固醇10 mg、姜黄素1 mg、人参皂苷Rb11 mg、超声间隔7 s。经验证GS-Rb1和Cur的包封率分别为(89.97±2.74)%和(90.98±1.32)%。体外细胞摄取实验结果表明,bEND.3和N2a细胞对MAN-Cur/GS-Rb1-Lips的摄取效率相比于Cur/GS-Rb1-Lips有明显提高。结论:成功优化用于制备MAN-Cur/GS-Rb1-Lips的处方,经MAN修饰的Cur/GS-Rb1-Lips,其脑靶向能力显著提高,被认为是一种很有潜力的脑靶向纳米给药系统。

Objective:To optimize the formulation of mannose(MAN)modified curcumin(Cur)/ginsenoside Rb1(GS-Rb1)co-loaded liposomes and investigate its targeting ability in vitro.Methods:Man-Cur/GS-Rb1 liposomes were prepared by a thin film dispersion ammonium sulfate gradient method.Taking the entrapment efficiency as the investigation index,the best formulation of MAN-Cur/GS-Rb1 liposomes was determined by central composite design response surface method.Active targeting of MAN-Cur/GS-Rb1-Lips in mouse brain microvascular endothelial cells(bEND.3)and mouse neuroblastoma cells(N2 a)was investigated by fluorescence microscopy.Results:Through Box-Behnken response surface method,the optimal formulation of liposomes was determined as DSPE-PEG 2000-MAN 2 mg,lecithin 88 mg,cholesterol 10 mg,curcumin 1 mg,ginsenoside Rb11 mg and ultrasonic interval 7 s.The entrapment efficiencies of ginsenoside Rb1 and curcumin were(89.97±2.74)%and(90.98±1.32)%,respectively.The results of cell uptake experiment in vitro showed that the uptake efficiency of MAN Cur/GS-Rb1 liposomes by bEND.3 and N2 a cells was significantly higher than that of Cur/GS-Rb1 liposomes.Conclusion:The formulation for the preparation of Man-Cur/GS-Rb1 liposomes is successfully optimized.MAN-modified Cur/GS-Rb1 liposomes have significantly improved brain targeting ability,and can be considered to be a potential brain targeted nano drug delivery system.