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中国药学(英文版) ›› 2016, Vol. 25 ›› Issue (9): 651-659.DOI: 10.5246/jcps.2016.09.072

• 【研究论文】 • 上一篇    下一篇

细胞内蛋白质在金纳米粒表面的吸附影响金纳米粒的细胞摄取和胞内分布

秦蒙蒙, 李轶凡, 何冰, 卫备, 代文兵, 张华, 王学清, 张强*   

  1. 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; 药学院 药剂学系, 北京 100191
  • 收稿日期:2016-04-07 修回日期:2016-05-03 出版日期:2016-09-28 发布日期:2016-05-15
  • 通讯作者: Tel.: +86-010-82802791, E-mail: zqdodo@bjmu.edu.cn
  • 基金资助:

    The National Basic Research Program of China (973 program, Grant No. 2015CB932100) & National Natural Science Foundation of China (Grant No. 81130059).

The adsorption of cellular proteins affects the uptake and cellular distribution of gold nanoparticles

Mengmeng Qin, Yifan Li, Bing He, Bei Wei, Wenbing Dai, Hua Zhang, Xueqing Wang, Qiang Zhang*   

  1. State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University Health Science Center, Beijing 100191, China
  • Received:2016-04-07 Revised:2016-05-03 Online:2016-09-28 Published:2016-05-15
  • Contact: Tel.: +86-010-82802791, E-mail: zqdodo@bjmu.edu.cn
  • Supported by:

    The National Basic Research Program of China (973 program, Grant No. 2015CB932100) & National Natural Science Foundation of China (Grant No. 81130059).

摘要:

纳米技术被广泛应用于医药领域, 其能够显著提高药物的生物利用度及靶向递送能力。然而, 纳米药物在体内环境中会吸附生物分子而影响其效能的发挥。已有的研究证明, 血浆中的蛋白质吸附在纳米粒表面后会改变纳米粒的表面性质。纳米粒在细胞转运过程中, 同样可能会吸附细胞中的某些蛋白。目前, 研究者们并不知道这些蛋白是否会影响纳米粒在下层细胞中的摄取。为了简化研究, 本研究将白蛋白包被的金纳米粒与Caco-2细胞裂解液共同孵育模拟纳米粒经上皮细胞转运过程; 利用TEM观察纳米粒与细胞裂解液孵育前后的表面形态; 使用SRB法评价孵育前后的纳米粒的细胞毒性; 通过ICP-MSCLSM实验测定金纳米粒在细胞中的摄取量和胞内分布情况。实验结果表明, 细胞蛋白的吸附能够增加金纳米粒在下层细胞的粘附和细胞摄取。裂解液孵育的金纳米粒和对照组纳米粒都主要分布在细胞的溶酶体中, 不同的是裂解液孵育的金纳米粒在线粒体有少量分布, 而对照组金纳米粒在线粒体中几乎没有分布。

关键词: 金纳米粒, 蛋白质, 细胞裂解液, 吸附, 促进, 细胞摄取

Abstract:

Nanotechnology has been widely used in the field of medicine, and it can significantly improve the bioavailability and the target efficiency of medicines. However, after administration, nanomedicines can adsorb biomolecules that can influence their effects. It was reported that the adsorption of plasma proteins can change the surface properties of nanoparticles. When nanoparticles pass through cells, they may carry some cellular proteins out of cells. Currently, it is unclear whether the adsorbed proteins affect the uptake of nanoparticles in the next cell layer. To simplify this complex biological process, BSA-capped gold nanoparticles were prepared and incubated with Caco-2 cell lysate to simulate conditions of transcytosis through epithelial cells. The surface morphology of nanoparticles was examined by TEM. SRB was used to evaluate the cytotoxicity of the nanoparticles. The uptake and cellular distribution of the nanoparticles were detected by ICP-MS and CLSM. The results suggested that the adsorption of cell proteins could enhance the adhesion and uptake of gold nanoparticles. The gold nanoparticles were mainly located in lysosomes, and there were some Lysate-capped AuNPs in the mitochondria whereas no BSA-capped AuNPs appeared there.

Key words: Gold nanoparticles, Proteins, Cell lysate, Adsorption, Enhance, Cellular uptake

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