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立方液晶纳米粒在MDCK-MDR1细胞模型中的摄取及摄取机制研究
谭宁, 杜守颖, 薛宇涛, 谭丽, 陆洋, 李鹏跃
北京中医药大学中药学院, 中药药剂学研究室, 北京 100102
摘要:
[目的]考察MDCK-MDR1细胞对立方液晶纳米粒的摄取及摄取机制。[方法]以钙黄绿素为标准荧光物质制备液晶纳米粒。采用流式细胞仪检测不同时间点MDCK-MDR1细胞内荧光强度,比较细胞对钙黄绿素、钙黄绿素液晶纳米粒摄取的差异;采用不同抑制剂(非律平、细胞松弛素D、氯丙嗪、2-D-去氧葡萄糖)与钙黄绿素液晶纳米粒共同孵育后,流式细胞仪测定胞内荧光强度,判断MDCK-MDR1细胞摄取液晶纳米粒的通路。[结果]摄取2 h内,立方液晶纳米粒不仅可以增加MDCK-MDR1细胞对钙黄绿素的摄取也可改变细胞的摄取行为。经氯丙嗪、2-D-去氧葡萄糖孵育的细胞胞内荧光含量低,差异有统计学意义(P<0.05)。[结论]立方液晶纳米粒可增加MDCK-MDR1对钙黄绿素的摄取,摄取途径为能量依赖网格蛋白介导的主动内吞。
关键词:  MDCK-MDR1  立方液晶纳米粒  摄取  机制  通路抑制剂
DOI:10.11656/j.issn.1672-1519.2018.01.15
分类号:R94
基金项目:北京市自然科学基金项目(7164274);2017年北京中医药大学研究生自主课题项目(2017-JYB-XS-072)。
Study on the cellular uptake and the up-take mechanism of cubic liquid crystalline nanoparticle on MDCK-MDR1 cells model
TAN Ning, DU Shouying, XUE Yutao, TAN Li, LU Yang, LI Pengyue
School of Chinese Material Medica, Beijing University of Chinese Medicine, Beijing 100102, China
Abstract:
[Objective] To study cellular uptake and pathway of cubic liquid crystalline nanoparticles (LCNPs) on MDCK-MDR1 cells.[Methods] Make LCNPs with calein (Cal-LCNPs), the standard fluorescent substance. Fluorescence intensity in MDCK-MDR1 cells was measured by flow cytometry at different time points. Different inhibitors (Filipin, Phytoalexins D, 2-Deoxy-D-glucose, chlorpromazine) were incubated with Cal-LCNPs. Flow cytometry was used to determine the intracellular fluorescence intensity then determine the pathways.[Results] Cubic liquid crystal nanoparticles can not only increase the uptake of Cal in MDCK-MDR1 cells in 2h, but also alter the uptake behavior of cells. MDCK-MDR1 uptakes Cal was consistent with the zero-order equation, and Cal-LCNPs' was in accordance with the first-order equation. The intracellular fluorescence of cells incubated with chlorpromazine and 2-D-deoxyglucose was low conpared with control (P<0.05).[Conclusion] Cubic liquid crystal nanoparticles can increase the uptake of calcein by MDCK-MDR1, and the pathway is energy-dependent clathrin mediated endocytosis initiative.
Key words:  MDCK-MDR1  LCNPs  uptake  mechanism  pathway inhibitor
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