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

[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(P0.05)。[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。

Study on the cellular uptake and the up-take mechanism of cubic liquid crystalline nanoparticle on MDCK-MDR1 cells model

[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.