乳铁蛋白修饰纳米脂质载体的制备及其脑靶向性评价

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目的选用乳铁蛋白(Lf)为脑靶向配体,构建受体介导的脑靶向姜黄素纳米脂质载体(Lf-Cur-NLCs),对其理化性质及体内脑靶向效率进行评价。方法采用熔融-乳化法制备姜黄素纳米脂质载体(Cur-NLCs),通过静电作用在姜黄素纳米脂质载体表面吸附乳铁蛋白,得到不同乳铁蛋白含量修饰的Lf-Cur-NLCs。考察其形态、粒径、Zeta电位、血浆稳定性及在含1%聚山梨酯80的生理盐水中的释放行为;选取乳铁蛋白质量浓度分别为0.5、1.5、2.0 mg·mL-1时制备的载荧光显像剂NIRD-15的乳铁蛋白修饰纳米脂质载体(分别标记为Lf1-NLC、Lf3-NLC和Lf4-NLC)进行小鼠尾静脉注射,采用活体成像系统观察小鼠活体及离体器官中药物荧光强度,评价Lf-NLCs的脑靶向性。结果姜黄素纳米脂质载体和乳铁蛋白姜黄素纳米脂质载体体系均呈类球形。姜黄素纳米脂质载体平均粒径(187.5±4.7)nm,Zeta电位(-23.7±2.9)mV。乳铁蛋白姜黄素纳米脂质载体体系平均粒径范围167.8~299.9 nm,Zeta电位的分布范围为-26.87~-13.03 mV。乳铁蛋白与姜黄素纳米脂质载体的静电吸附作用存在一个吸附与解吸附的过程,当乳铁蛋白浓度为2.0 mg·mL-1,温孵时间为6 h时,乳铁蛋白在姜黄素纳米脂质载体表面的吸附趋于饱和。乳铁蛋白姜黄素纳米脂质载体在血浆中具有较好的稳定性,体外释放具有明显的缓释特征。与NLCs相比,尾静脉注射5 min后,姜黄素纳米脂质载体在脑内有较强的荧光,说明姜黄素纳米脂质载体能主动靶向脑组织,同时研究发现Lf3-NLC的脑靶向效果最好。结论本实验利用静电吸附作用成功构建了具有脑靶向功能的姜黄素纳米脂质载体,避免了靶向载体设计中的化学合成过程,工艺简单,具有较好的发展前景,但载体脑靶向能力与乳铁蛋白的用量有关。 OBJECTIVE: To evaluate the physical and chemical properties of brain-targeted curcumin-loaded N-liposomes (Lf-Cur-NLCs) and their brain targeting efficiency by using Lf as brain targeting ligand. . Methods Cur-NLCs were prepared by melt-emulsification method. Lf-Cur-NLCs with different contents of lactoferrin were obtained by electrostatic adsorption of lactoferrin on curcumin nanoparticles. The morphology, particle size, Zeta potential, plasma stability and release behavior in 1% polysorbate 80 normal saline were investigated. When the concentration of lactoferrin was 0.5,1.5,2.0 mg · mL-1 (Lf1-NLC, Lf3-NLC and Lf4-NLC) containing the fluorescent imaging agent NIRD-15 were injected into the tail vein of mice, and the in vivo imaging system was used to observe the mice in vivo and in vivo Fluorescence intensity of isolated drug in vivo to evaluate brain targeting of Lf-NLCs. Results Both curcumin nanoliposomes and lactoferricin curcumin nanosliposomes were spheroidal. The average diameter of curcumin nanofiber carrier (187.5 ± 4.7) nm, Zeta potential (-23.7 ± 2.9) mV. Lactoferrin curcumin nanosized lipid carrier system average particle size range of 167.8 ~ 299.9 nm, Zeta potential distribution range of -26.87 ~ -13.03 mV. There was a process of adsorption and desorption of lactoferrin and curcumin nanosliposomes. When lactoferrin concentration was 2.0 mg · mL-1 and incubation time was 6 h, The adsorption on the surface of the nanosized lipid carrier tends to be saturated. Lactoferrin curcumin nanoliposomes have good stability in plasma and release in vitro has obvious sustained-release characteristics. Compared with NLCs, 5 min after tail vein injection, curcumin nanoliposomes had a strong fluorescence in the brain, indicating that curcumin nanoliposomes can actively target brain tissue. At the same time, it was found that brain targets of Lf3-NLC To the best effect. Conclusion In this experiment, curcumin nanoliposomes with brain-targeting function were successfully constructed by electrostatic adsorption, avoiding the chemical synthesis process in targeting vector design. The process was simple and had a good prospect. However, Capacity and lactoferrin dosage related.
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