APP/PS1小鼠中PEG-PLA纳米粒的表面蛋白冠组成及其脑内递送的实验研究

摘要 目的：研究阿尔茨海默病（Alzhemer''s disease，AD）模型鼠中聚乙二醇聚乳酸（poly(ethylene glycol)-poly(l-lactide)，PEG-PLA）纳米粒表面蛋白冠组成及其对脑内递送特性的影响。方法：制备PEG-PLA纳米粒，测定纳米粒的zeta电位及粒径，采用透射电子显微镜观察纳米粒形态。通过双光子显微镜观察APP/PS1小鼠与野生型（Wild Type，WT）小鼠脑内PEG-PLA纳米粒分布特性。采用液相色谱-质谱联用（LC-MS）技术对PEG-PLA纳米粒分别与APP/PS1小鼠和WT小鼠血浆孵育形成的两种不同蛋白冠进行蛋白组学分析。结果：制备的PEG-PLA纳米粒粒径均一，分散性较好。静脉注射PEG-PLA后，APP/PS1小鼠脑内纳米粒量明显高于WT小鼠。蛋白质组学结果显示，APP/PS1小鼠血浆孵育组PEG-PLA纳米粒表面蛋白冠中凝聚素（Clusterin）明显高于WT小鼠血浆孵育组，该蛋白与纳米粒逃避机体清除有关。此外，纳米粒蛋白冠中血管性血友病因子（Von Willebrand factor）、玻连蛋白（Vitronectin）、肌球蛋白重链-9（Myosin-9）等参与细胞粘附作用相关蛋白在APP/PS1小鼠血浆孵育组也明显多于WT小鼠血浆孵育组。结论：PEG-PLA纳米粒在AD模型小鼠中表现出的高入脑量，可能与AD疾病影响纳米粒蛋白冠组成有关。

The Study of Protein Corona Composition and Brain Delivery of PEG-PLA Nanoparticles in APP/PS1 Mice

ABSTRACT Objective: The aim of this study is to investigate the protein corona composition and the brain delivery of poly(ethylene glycol)-poly(l-lactide) (PEG-PLA) nanoparticles in Alzhemer''s disease (AD) model mice. Methods: PEG-PLA nanoparticles were prepared. The particle size and zeta potential of the nanoparticles were measured, and their morphology were observed under transmission electron microscope. The distribution of PEG-PLA nanoparticles in the brain of APP/PS1 mice and wild type (WT) mice were observed under two-photon microscopy. Composition of the two different protein coronas on nanoparticles incubated in the APP/PS1 mice plasma and WT mice plasma were analyzed by LC-MS. Results: The prepared PEG-PLA nanoparticles exhibited uniform size and good dispersion. After intravenously administered with PEG-PLA nanoparticles, APP/PS1 mice showed higher content of nanoparticles in brain than that of WT mice. Proteomic analysis showed that clusterin in the protein corona formed on PEG-PLA nanoparticles in APP/PS1 mice plasma incubation group were much higher than that of WT mice plasma incubation group. Clusterin is related to the evasion of clearance of nanoparticles in vivo. In addition, some corona components related to cell adhesion such as von Willebrand factor, vitronectin and myosin-9 were higher in APP/PS1 mice plasma incubation group compared to WT mice plasma incubation group. Conclusion: AD model mice showed high content of PEG-PLA nanoparticles in the brain. This may be related to the protein corona composition influenced by AD state.