去唾液酸糖蛋白受体介导的肝靶向纳米脂质超声造影剂的制备及体外实验

目的 以肝细胞膜特异性受体去唾液酸糖蛋白受体为靶向受体,利用共价结合的原理,制备出肝靶向性液态氟碳纳米超声造影剂,观察该造影剂的一般特性、与人肝细胞L02的靶向结合及体外聚集显像效果.方法 利用还原胺法制备去唾液酸糖蛋白特异性配体半乳糖化多聚赖氨酸(Gal-PLL);利用旋转蒸发及声振法制备液态氟碳纳米脂质超声造影剂;培养人肝细胞L02,于不同时间点观察与L02细胞的靶向结合;Philips iU 22超声诊断仪L12-5探头对比观察靶向液态氟碳纳米脂质超声造影剂的体外显像效果.结果 靶向液态氟碳纳米脂质超声造影剂粒径极小,分布均匀,形态呈圆球形,且能与L02有效结合;体外乳胶囊显示:脱气水侧呈现无回声,靶向液态氟碳纳米脂质超声造影剂侧呈现高回声.结论 以去唾液酸糖蛋白受体为靶向受体,自制的携半乳糖化多聚赖氨酸的靶向液态氟碳纳米脂质超声造影剂能有效与人肝细胞LO2靶向结合,该靶向超声造影剂能在体外聚集显影.该靶向超声造影剂粒径极小,是细胞水平上肝靶向性超声分子显像的一种理想的超声分子探针.

Abstract：

Objective To prepare the liver targeting nano-liquid perfluorocarbon ultrasound contrast agent and observe its general characteristics;to observe the targeting combined effects of the human liver cells L02 and the targeted ultrasound contrast agent ;to evaluate the gathering imaging effects of the targeted ultrasound contrast agent. Methods Amine method was used to prepared asialoglycoprotein Gal specific ligand polylysine (Gal-PLL), rotary evaporator and sonicated liquid fluorocarbon were used to prepare nano lipid ultrasound contrast agent. Human liver cell L02 were cultured, the combined effects were observed according to the reacting time of the cells and the targeted nano-lipid ultrasound contrast agent. The nanolipid ultrasound contrast agent and the degassed_ water_ were loaded into cysts and their ultrasound imaging effects were observed by ultrasound diagnostic apparatus Philips iU22. Results The particle size of the liquid fluorocarbon nano-targeted lipid ultrasound contrast agent was extremely small, uniform, cylindrical and spherical. The cysts in vitro showed that the side of the targeted liquid perfluorocarbon nano-lipid ultrasound contrast agent showed high echo. Conclusions The targeted liquid perfluorocarbon nano-lipid ultrasound contrast agent can be effectively targeted to the human liver cells L02 due to carrying home-made Gal-PLL. The targeted ultrasound contrast agents can be imaging by ultrasound and be confirmed in vitro.The size of the contrast agent was small, therefore, it can be considered an ideal ultrasonic molecular probe and achieve the ultrasound molecular imaging in cell level.

Preparation of asialoglycoprotein receptor-mediated hepatic targeting nano- lipid ultrasound contrast agent and its ultrasound imaging in vitro

Objective To prepare the liver targeting nano-liquid perfluorocarbon ultrasound contrast agent and observe its general characteristics;to observe the targeting combined effects of the human liver cells L02 and the targeted ultrasound contrast agent ;to evaluate the gathering imaging effects of the targeted ultrasound contrast agent. Methods Amine method was used to prepared asialoglycoprotein Gal specific ligand polylysine (Gal-PLL), rotary evaporator and sonicated liquid fluorocarbon were used to prepare nano lipid ultrasound contrast agent. Human liver cell L02 were cultured, the combined effects were observed according to the reacting time of the cells and the targeted nano-lipid ultrasound contrast agent. The nanolipid ultrasound contrast agent and the degassed_ water_ were loaded into cysts and their ultrasound imaging effects were observed by ultrasound diagnostic apparatus Philips iU22. Results The particle size of the liquid fluorocarbon nano-targeted lipid ultrasound contrast agent was extremely small, uniform, cylindrical and spherical. The cysts in vitro showed that the side of the targeted liquid perfluorocarbon nano-lipid ultrasound contrast agent showed high echo. Conclusions The targeted liquid perfluorocarbon nano-lipid ultrasound contrast agent can be effectively targeted to the human liver cells L02 due to carrying home-made Gal-PLL. The targeted ultrasound contrast agents can be imaging by ultrasound and be confirmed in vitro.The size of the contrast agent was small, therefore, it can be considered an ideal ultrasonic molecular probe and achieve the ultrasound molecular imaging in cell level.