硅质体的形状对其跨Caco-2细胞单层转运的影响  

The effect of the shape of nanocarriers on their transmembrane across Caco-2 cell monolayer

作　　者：张润宇 宋阳 陈晴 邓海亮 张华[1] 代文兵[1] 何冰[1] 陈英[2] 张强[1] 王学清[1] ZHANG Run-yu;SONG Yang;CHEN Qing;DENG Hai-liang;ZHANG Hua;DAI Wen-bing;HE Bing;CHEN Ying;ZHANG Qiang;WANG Xue-qing(School of Pharmaceutical Sciences,Peking University,Beijing 100191,China;Guangdong Institute for Drug Control,Guangzhou 510700,China)

机构地区：[1]北京大学药学院,北京100191 [2]广东省药品检验所,广东广州510700

出　　处：《药学学报》2020年第8期1932-1940,共9页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金面上项目(81872809,31671017)。

摘　　要：本文旨在构建球形和盘状两种不同形状的硅质体,以研究纳米载体的形状对其细胞摄取及跨膜能力的影响。经卤代、亲核加成及酰化反应合成硅质体复合脂质(cerasome-forming lipid,CFL),并利用质谱和核磁共振氢谱进行鉴定。将CFL与短链脂材1,2-dihexanoyl-sn-glycero-3-phosphocholine(DHPC)通过薄膜水化法制备有机-无机杂化的纳米盘(nanodisc,简称纳米盘),并使用CFL制备球形硅质体(nanosphere,简称硅质体)。用激光粒度仪测定硅质体和纳米盘的粒径及电位,透射电镜观察其形状。以人源结肠癌Caco-2细胞为模型,采用激光共聚焦显微镜定性考察形状对纳米载体在细胞上摄取和转运的影响,采用高效液相色谱法定量考察形状对其跨膜量的影响。结果表明:制得的硅质体及纳米盘粒径相近,均约为110 nm,电位约为-25 mV,在电镜下形状规则均一。在20 min内,纳米盘的入胞速率显著快于硅质体,进一步在Caco-2细胞单层模型上的研究结果表明,摄取更快的纳米盘在细胞单层中累积量更少,即更快地被转运出胞,在2 h内纳米盘的跨膜量均高于硅质体。该结果提示通过合理设计纳米载体的形状可以调控纳米-细胞的相互作用,促进纳米载体的跨膜转运,增加药物的吸收。Cerasomes with different shapes were constructed to investigate the effect of the nanocarriers’shape on the cellular uptake and transmembrane capacity.Cerasome-forming lipid(CFL)was synthesized via halogenation,nucleophilic addition and acylation reaction and detected by mass spectrometry and nuclear magnetic resonance spectroscopy.CFL and short chain 1,2-dihexanoyl-sn-glycero-3-phosphocholine(DHPC)were employed to prepare organic-inorganic hybrid bicelles in discal shapes(nanodisc)by the thin-film hydration method,and CFL was also used to prepare spherical cerasomes(nanosphere).The particle size and zeta potential of nanocarriers were measured by dynamic light scattering analysis,and the morphology was observed by transmission electron microscopy.With human colon cancer cell line Caco-2 as the model,the effect of the shape of nanocarriers on cellular uptake and transmembrane capacity was investigated qualitatively by confocal laser scanning microscope(CLSM),and the transmembrane capacity was analyzed quantitatively by high performance liquid chromatography(HPLC).The results showed that nanosphere and nanodisc had similar particle diameters around 110 nm and similar zeta potential around-25 mV,with regular morphology under transmission electron microscope.The cellular uptake rate of nanodisc was significantly higher than that of nanosphere in 20 minutes.Further research on Caco-2 cell monolayer demonstrated that nanodisc with faster uptake had less accumulation in the monolayer,which means it had a higher transmembrane rate on Caco-2 cell monolayer and the transmembrane capacity of the nanodisc was better than that of nanosphere within 2 h.These results suggest that rational design of the shape of nanocarriers is expected to regulate nano-bio interactions,promote the transmembrane transport of nanocarriers,and improve the drug absorption.

关 键 词：硅质体 纳米盘 形状 CACO-2细胞单层 跨膜转运 

分 类 号：R943[医药卫生—药剂学]