金纳米粒用于口服内吞转运机制研究的检测方法

doi:10.5246/jcps.2016.10.082

中国药学(英文版) ›› 2016, Vol. 25 ›› Issue (10): 737-746.DOI: 10.5246/jcps.2016.10.082

金纳米粒用于口服内吞转运机制研究的检测方法

刘德春¹, 杨丹¹, 何冰², 代文兵², 张华², 王学清², 袁兰^3*, 张强^2*, 唐星^1*

1. 沈阳药科大学药学院, 辽宁沈阳 110016
2. 北京大学医学部药学院天然药物及仿生药物国家重点实验室, 北京 100191
3. 北京大学医学部医药卫生分析中心, 北京 100191

收稿日期:2016-05-16 修回日期:2016-06-20 出版日期:2016-10-27 发布日期:2016-07-10
通讯作者: Tel./Fax: +86-024-23986343, +86-10-82802791, E-mail: xiaoqi_16@126.com, zqdodo@bjmu.edu.cn
基金资助:
The National Basic Research Program of China (973 program, Grant No. 2015CB932100) and the National Natural Fund Project (Grant No. 91330103).

A practical method to trace intracellular and transcellular transport pathways of gold nanoparticles via oral administration

Dechun Liu¹, Dan Yang¹, Bing He², Wenbing Dai², Hua Zhang², Xueqing Wang², Lan Yuan^3*, Qiang Zhang^2*, Xing Tang^1*

1. Department of Pharmaceutics, College of Pharmacy, Shenyang Pharmaceutical University, Shenyang 110016, China
2. State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University Health Science Center, Beijing 100191, China
3. Medical and Healthy Analytical Center, Peking University Health Science Center, Beijing 100191, China

Received:2016-05-16 Revised:2016-06-20 Online:2016-10-27 Published:2016-07-10
Contact: Tel./Fax: +86-024-23986343, +86-10-82802791, E-mail: xiaoqi_16@126.com, zqdodo@bjmu.edu.cn
Supported by:
The National Basic Research Program of China (973 program, Grant No. 2015CB932100) and the National Natural Fund Project (Grant No. 91330103).

摘要/Abstract

摘要：

口服纳米载体可以有效地提高难溶性药物的生物利用度, 其与肠上皮细胞的相互作用研究也成为一大热点。在该领域的研究中, 物理包载或化学偶联荧光基团的纳米载体常常被选择, 但是物理包载荧光染料可能会发生泄漏, 影响结果分析, 而化学偶联荧光探针可能会改变纳米粒结构的表面特征。因此, 需要寻找一种简便、准确的检测方法以探究口服纳米粒内吞及跨膜转运的过程、胞内分布和机制。本文选择金纳米粒为纳米模型载体, 基于金纳米粒具有光散射的性质, 以633 nm波长的激光为激发光源, 通过激光扫描共焦显微镜(CLSM)检测金纳米粒溶液, 肠道上皮细胞内及大鼠肠道中的金纳米粒。结果通过激光共聚焦显微镜反射光的模式, 无论在细胞水平还是大鼠肠道中, 都能够准确地检测到金纳米粒的信号, 进而分析金纳米粒入胞及跨越肠道上皮细胞的过程和胞内分布, 且不受背景的干扰, 并能借助层扫模式, 构建金纳米粒在细胞内分布的3D模型。因此, 在不进行荧光探针偶联的条件下, 通过反射光分析金纳米粒跨膜转运是准确、简便的方法。

关键词: 金纳米粒, 反射光, 激光扫描共焦显微镜, 口服给药, 内吞和跨膜转运, 体内

Abstract:

Oral nanoparticles play an important role in improving the bioavailability of poorly water-soluble drug. It is necessary to investigate the interaction of nanoparticles with intestinal epithelial cells. In general, nano-carriers labeled with fluorescent probes are always chosen. However, fluorescent dye via physical loading may leak in complex biological environment and lose its function to trace the transport behavior of nanoparticles. Fluorescent probes chemically coupled on the nanoparticles may alter the properties of nanoparticles. Therefore, a facile and exact detection method is required to trace intracellular and transcellular pathways of oral nano-medicines. In our study, gold nanoparticles were selected as nano-carriers owing to their unique characteristics of light scattering. The feasibility of gold nanoparticle detection through reflected light signal was tested in different situations, including gold nanoparticle solution, cell and animal level. As a result, high resolution image of gold nanoparticles could be detected through reflection mode by confocal laser scanning microscope (CLSM) when excited at a wavelength of 633 nm. The reflected light signal of gold nanoparticles could be clearly shown in different intestinal epithelial cells no matter when they were in fixed or in living state, and the intracellular trafficking and distribution of gold nanoparticles were clearly shown in three-dimensional image. Meanwhile, this method was also applied to rat small intestine in vivo. In conclusion, we believed that this technique was a convenient and precise way to explore the transport behavior of gold nanoparticles via oral administration without fluorescent dye.

Key words: Gold nanoparticles, Reflected light signal, CLSM, Oral administration, Intracellular and transcellular pathways, In vivo

中图分类号:

R943

Supporting:

刘德春, 杨丹, 何冰, 代文兵, 张华, 王学清, 袁兰, 张强, 唐星. 金纳米粒用于口服内吞转运机制研究的检测方法[J]. 中国药学(英文版), 2016, 25(10): 737-746.

Dechun Liu, Dan Yang, Bing He, Wenbing Dai, Hua Zhang, Xueqing Wang, Lan Yuan, Qiang Zhang, Xing Tang. A practical method to trace intracellular and transcellular transport pathways of gold nanoparticles via oral administration[J]. Journal of Chinese Pharmaceutical Sciences, 2016, 25(10): 737-746.

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金纳米粒用于口服内吞转运机制研究的检测方法

A practical method to trace intracellular and transcellular transport pathways of gold nanoparticles via oral administration

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