Uptake of albumin nanoparticle surface modified with glycyrrhizin by primary cultured rat hepatocytes

AIM: To investigate the uptake difference between bovine serum albumin nanoparticle (BSA-NP) and bovine serum albumin nanoparticles with their surface modified byglycyrrhizin (BSA-NP-GL) and to develop a novel hepatocyte targeting BSA-NP-GL based on active targeting technology mediated by specific binding site of GL on rat cellular membrane. METHODS: Calcein loaded bovine serum albumin nanoparticles (Cal-BSA-NP) were prepared by desolvation process. Glycyrrhizin was conjugated to the surface reactive amino groups (SRAG) of Cal-BSA-NP by sodium periodate oxidization, which resulted in calcein-loaded bovine serum albumin nanoparticles with their surface modified by glycyrrhizin (Cal-BSA-NP-GL). The morphology of the two types of prepared nanoparticles (NP) was observed by transmission electron microscopy. The diameter of NP was measured with a laser particle size analyzer. The interaction between Cal-BSA-NP-GL and primary cultured hepatocytes was studied through cellular uptake experiments. The uptake amount of Cal-BSA-NPGL and Cal-BSA-NP by rat hepatocytes was determinedby fluorospectrophotometry. Uptake characteristics were investigated through experiments of competitive inhibition of specific binding site of GL. RESULTS: Both Cal-BSA-NP-GL and Cal-BSA-NP had regular spherical surfaces. The average diameter of CalBSA-NP-GL and Cal-BSA-NP was 77 and 79 nm respectively. The uptake amount of the two NP by hepatocytes reached its maximum at 2 h after incubation. The uptake amount of Cal-BSA-NP-GL by rat hepatocytes was 4.43-fold higher than that of Cal-BSA-NP. There was a significant difference in the uptake of Cal-BSA-NP-GL and Cal-BSA-NP by hepatocytes (P＜0.01). The uptake of Cal-BSA-NP-GL was inhibited when GL was added previously to isolated rat hepatocytes, and the uptake of Cal-BSA-NP was not affected by GL.CONCLUSION: A binding site of GL is present on the surface of rat hepatocytes, BSA-NP-GL may be internalized via this site by hepatocytes and can be used as a drug carrier for active targeting of delivery drugs to hepatocytes.

甘草酸表面修饰万乃洛韦白蛋白纳米粒的制备及其肝靶向性研究

以万乃洛韦为模型药物 ,去溶剂化法制备普通载药纳米粒 ,结合高碘酸盐氧化法制备甘草酸 -万乃洛韦白蛋白纳米粒偶联物。对其表面甘草酸密度、形态、大小及其分布、体外释药特性、载药量、包封率、动物体内肝分布和体外肝细胞的摄取情况进行了研究。修饰纳米粒表面甘草酸密度为 9;平均粒径 d0 .5=2 6 8± 2 3nm;载药量1.35 % ;包封率 6 8.76 % ;体外释药符合双相动力学规律 ;对肝细胞具有选择靶向性。静注 15 min后 ,有 6 9.89%集中在肝脏 ,对照组为 6 4 .82 % ,二者之间存在显著差异 (P<0 .10 )。甘草酸表面修饰白蛋白纳米粒制备成功 ,为肝细胞靶向给药提供了新途径。

肝细胞靶向甘草酸表面修饰白蛋白纳米粒的制备工艺

目的 研究能主动靶向于肝实质细胞的甘草酸表面修饰白蛋白纳米粒的制备工艺。方法 去溶剂化法制备普通纳米粒 ,以高碘酸盐氧化法制备甘草酸 白蛋白纳米粒偶联物。用 2 ,4,6 三硝基苯磺酸显色法与凝胶柱色谱法验证是否偶联成功 ,HPLC法测定修饰纳米粒表面甘草酸密度。结果 修饰纳米粒表面活性氨基数量较对照组减少 1 9 6 % ;偶联于纳米粒表面的甘草酸密度为 9 2 %。纳米粒形态圆整 ,平均粒径为 73nm ,在 2 5℃和 37℃条件下放置 1 0d后 ,性质稳定。结论 甘草酸表面修饰白蛋白纳米粒制备成功 。