北京生物医学工程

二巯基丁二酸修饰的Fe3O4纳米颗粒对人血管内皮细胞的作用研究

Study on the interactions of dimercaptosuccinic acid modified magnetite nanoparticles to human vascular endothelial cells

作者：杜丽帆张宇杨爱云温涛刘健孟洁许海燕

单位：中国医学科学院基础医学研究所，北京协和医学院基础学院（北京 100005）东南大学生物科学与医学工程学院，江苏省生物材料与器件重点实验室（南京 210096）

关键词： Fe3O4纳米颗粒；内皮细胞；吞噬；血管内皮生长因子

分类号：R318.08

出版年·卷·期（页码）：2019·38·3（221-226）

摘要：

目的研究二巯基丁二酸修饰的Fe3O4纳米颗粒（dimercaptosuccinic acid-magnetite nanoparticles，DMSA-Fe3O4）对人脐静脉内皮细胞（human umbilical vein endothelial cells, HUVECs）功能的影响。方法利用动态光散射法表征DMSA-Fe3O4的粒径及表面电荷；采用普鲁士蓝染色、邻二氮菲铁定量和透射电镜观察方法研究HUVECs对DMSA-Fe3O4的摄取规律；利用细胞计数试剂盒（Cell Counting Kit-8, CCK-8）检测DMSA-Fe3O4对内皮细胞活性的影响；通过酶联免疫吸附试剂盒测定DMSA-Fe3O4对内皮细胞血管内皮生长因子（vascular endothelial growth factor, VEGF）分泌量的影响。结果 HUVECs能够大量吞噬DMSA-Fe3O4，其吞噬量具有孵育时间和剂量依赖性；短时间内所测剂量范围对细胞活性无显著影响，但长时间高剂量条件使细胞活性明显降低。此外，在高剂量DMSA-Fe3O4暴露下（200 μg/mL），内皮细胞分泌VEGF的量约为对照组的3倍。结论 DMSA-Fe3O4易于被HUVECs吞噬；高浓度DMSA-Fe3O4与HUVECs长时间培养使细胞活性降低，并刺激内皮细胞分泌VEGF。

Objective To investigate the interactions between dimercaptosuccinic acid modified Fe3O4 nanoparticles (DMSA-Fe3O4) and human umbilical vein endothelial cells (HUVECs). Methods The particle size distribution and Zeta potential of DMSA-Fe3O4 were analyzed by dynamic light scattering. The cellular uptake of DMSA-Fe3O4 by HUVECs was measured with Prussian blue staining, phenanthroline and transmission electron microscopy observation. Cell Counting Kit-8 kit was used to determine the viability of DMSA-Fe3O4 treated endothelial cells. The secretion of vascular endothelial growth factor (VEGF) by the cells was quantifed by enzyme-linked immunosorbent assay kit. Results Agglomerate nanoparticles were observed in the HUVECs cytoplasm, the cellular uptake of DMSA-Fe3O4 was time and dose dependent. The viability of HUVECs was not affected with 6-h incubation of DMSA-Fe3O4, while significantly decreased at 24-h and 48-h incubation. In addition, the endothelial cells secreted higher level of VEGF due to injury stress mediated by DMSA-Fe3O4. Conclusions DMSA-Fe3O4 was easily uptaken by HUVECs, which inhibited the cell viability in high dose and long term, meanwhile induced HUVECs secret VEGF.

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