0X26偶联载内吗啡肽超支化聚甘油－聚乳酸－聚乙醇酸纳米粒透血脑屏障机制实验研究  

Preparation of brain active-targeting endomorphin loaded nanoparticles and study on its effects of passing across blood brain barrier

作　　者：金旭[1] 熊蔚[1] 曾敏[1] 吕丰[2] 包汉梅[2] 刘天军[2] 

机构地区：[1]首都医科大学附属北京天坛医院麻醉科,北京100050 [2]中国医学科学院北京协和医学院生物医学工程研究所,天津300192

出　　处：《国际生物医学工程杂志》2014年第3期135-138,I0003,共5页International Journal of Biomedical Engineering

基　　金：国家自然科学基金资助项目（30700780）;北京市自然科学基金资助项目（7102052）;北京市科技新星计划项目（2008A083）;北京市卫生系统高层次卫生技术人才项目（学科骨干）（2013-3-047）;天津市生物医学材料重点实验室开放课题项目（2013-1-008）

摘　　要：目的 制备新型包载内吗啡肽（EM）的偶联脑主动靶向抗体OX26的超支化聚甘油-聚乳酸-聚乙醇酸（HBPG-PLGA）纳米粒,通过脑毛细血管内皮细胞（BMEC）研究其透血脑屏障（BBB）机制.方法 利用新型材料HBPG-PLGA通过复合乳液法制备载EM的纳米粒,在其表面偶联转铁蛋白受体单克隆抗体OX26,通过透射电镜等进行表征;采用流式细胞术和激光共聚焦显微镜研究不同配方纳米粒透过BBB的机制.结果 制备的载EM HBPG-PLGA纳米粒稳定,电镜下呈核壳结构,平均粒径（170±20） nm,Zeta电位约-27 mV,与OX26抗体的偶联效果较好,药物可以缓慢释放72 h以上.流式细胞术结果表明载EMHBPG-PLGA纳米粒具有良好的透BBB能力,其机制与细胞内吞作用有关;激光共聚焦显微镜结果显示纳米粒被内吞后,EM与纳米材料在细胞内分离,EM进入BBB,产生药理作用.结论 制备的偶联OX26载EM纳米粒稳定,能达到较好的透BBB作用,其机制与BMEC内吞作用有关,内吞后药物分离,进入脑内产生药理作用.Objective To prepare a novel brain active-targeting endomorphin （EM） loaded hyperbranched polyglycerols-poly （lactic-co-glycolic acid） （HBPG-PLGA） nanoparticles （NPs） and study its mechanism of passing across blood brain barrier （BBB） in brain microvascular endothelial cells （BMEC）.Methods The OX26 （transferring receptor monoclonal antibody） conjugated EM loaded HBPG-PLGA NPs was constructed according to water-in-oil-in-water emulation solvent evaporation technique as a novel biodegradable brain active-targeting drug delivery system.The properties of the NPs were evaluated by transmission electron microscope （TEM） in vitro.Through flow cytometry and laser scanning confocal microscope,the mechanism of passing across BBB was evaluated.Results The preparation methodology of NPs was optimized and established.The mean diameter was （170±20） nm and Zeta potential was about-27 mV.Core-shell construction was showed on TEM.Cellular uptake study showed that the uptake of NPs was via a caveolae-mediated endocytic pathway,then endomorphin and carrier were divided into two parts in BMEC.Conclusions The OX26 conjugated EM loaded NPs were stable,and demonstrate remarkable effects on crossing BBB.Cellular uptake by BMEC is a very important mechanism of the NPs＇ brain activating-targeting effect.

关 键 词：纳米粒 主动靶向 疼痛治疗 血脑屏障 内吞 

分 类 号：R318[医药卫生—生物医学工程] TB383[医药卫生—基础医学]