OX26/ApoE共修饰脑靶向载黄连解毒汤介孔硅纳米载体的构建及治疗脑缺血再灌注损伤的PK/PD研究

Ischemic cerebrovascular disease, well known with its high incidence, high death rate, high relapse rate and high mutilation rate, becomes the major pathogenic factor in cerebral apoplexy and causes serious harm to human health. Recent years, compound preparations of Traditional Chinese Medicine (TCM) receive more attentions for their unique effects in preventing and treating central nervous system diseases and lower side effects. The classical prescription Huang-Lian-Jie-Du Decoction is an outstanding representative of them for satisfactory curative effect on treating cerebral ischemia reperfusion injury. However the unneglected limitations of TCM seriously restrict the clinical applications, such as high dosage in single administration, bad patient compliance, low penetration through BBB, and so on. Thus the urgent problem now confronting us is to construct a brain-targeted drug deliver system with high loading capacity and specifically targeted profile loading Huang-Lian-Jie-Du Decoction active fraction. In this study, mesoporous silica nanoparticles (MSNs) with high loading capacity and good drug release profile is coupled with ligands OX26 and ApoE to prepare modified nanoparticles OX26-ApoE-MSNs, which are used for loading Huang-Lian-Jie-Du Decoction active fraction. The OX26-ApoE-MSNs penetrate through BBB by OX26 and ApoE receptor mediated endocytosis and increase the biological utilization rate by accumulating nanoparticles in brain. BBB model is established by brain endothelial cell lines bEnd.3, the penetration mechanism through BBB are investigated by Ussing Chamber experiment, and the pharmacokinetics-pharmacodynamics correlation analysis is applied to evaluate the protecting effect on cerebral ischemia reperfusion injury. By building a brain-targeted drug deliver system with high loading capacity and specifically targeted profile, the research will provide new methods and ideas in the modernization of TCM.

缺血性脑血管疾病是中老年人的一种常见病、多发病，是脑卒中发病的主要因素，严重危害人类健康。中药复方制剂在预防和治疗中枢神经系统疾病方面有独特疗效。研究表明，黄连解毒汤预防及治疗脑缺血再灌注损伤时疗效确切。但传统中药汤剂在临床治疗脑血管疾病时具有单次给药剂量大，耐受性差，BBB透过能力低等局限。如何实现黄连解毒汤有效部位的高效包载及靶向治疗是急需解决的问题。本研究拟合成具有高载药量和良好释药特性的介孔硅纳米粒，通过与OX26/ApoE偶联，得到黄连解毒汤有效部位脑靶向介孔硅纳米载体，实现脑部定位治疗，提高药物的脑内生物利用度；采用bEnd.3细胞建立BBB模型，运用Ussing Chamber技术阐明该纳米载体透过BBB的机制；运用PK/PD技术研究该载药体系对脑缺血再灌注损伤的保护作用。本研究将建立一种高载药量、高靶向性的脑部给药系统，为经典复方中药的现代靶向制剂的研究提供新方法和思路。

缺血性脑血管疾病是中老年人的一种常见病、多发病，是脑卒中发病的主要因素，严重危害人类健康。以黄连解毒汤为代表的中药复方制剂在预防和治疗中枢神经系统疾病方面有独特疗效。本研究以介孔硅为载体，成功包载黄连解毒汤，通过与OX26/ApoE偶联制备了脑靶向纳米载体；体外细胞实验表明，OX26/ApoE修饰的介孔硅纳米粒被小鼠脑微血管内皮细胞bEnd.3的摄入量明显增加，表明介孔硅纳米粒经过配体修饰后，能够增加脑微血管内皮细胞对其的摄取，脑靶向性明显增强；体内组织分布实验表明，介孔硅纳米粒能够延长在体内的滞留时间，经OX26/ApoE修饰后能明显提高在脑内的分布；实验中建立了大鼠脑缺血再灌注模型，以神经功能学评分、脑梗死体积百分率、脑组织病理形态学变化作为指标，对药效学结果进行评价，实验表明OX26/ApoE修饰的黄连解毒汤介孔硅纳米粒能够有效改善脑缺血再灌注损伤模型大鼠神经功能缺陷，脑梗死面积减小，大鼠脑组织海马区神经元结构明显改善，能显著提高治疗局造性脑缺血再灌注损伤的效果。本研究建立了一种高载药量、高靶向性的脑部给药系统，通过本课题的实施，为经典复方中药的现代靶向控释制剂的研究提供新方法和思路。本课题共发表SCI论文2篇，中文核心论文2篇，培养博士后1名，博士研究生1名，硕士研究生5名。