蟾毒灵纳米混悬液对HepG2细胞增殖及细胞摄取研究

Study of Bufalin Nanosuspensions on Proliferation and Cellular Uptake of HepG2 Cells

目的：制备蟾毒灵纳米混悬液（BU-NS）并研究其制剂学特点,以及该制剂对人肝癌HepG2细胞增殖及细胞摄取的影响。方法：采用反溶剂沉淀法制备BU-NS。用粒度仪测定其粒径,用透射电镜（TEM）进行形态学考察。采用MTT法检测蟾毒灵原药（BU）和BU-NS对HepG2细胞的增殖抑制作用,运用高效液相色谱法（HPLC）对药物细胞摄取进行定量研究。结果：BU-NS的平均粒径为（390.0±8.1）nm,多分散系数为0.22±0.04,Zeta电位为（-9.05±1.21）mV。TEM观察发现BU-NS呈不规则球形,粒径与粒度仪所测结果基本一致。短期物理稳定性研究表明,BU-NS在4℃下避光保存15 d后基本保持稳定。细胞增殖抑制作用实验表明,4 h内BU-NS对HepG2细胞增殖的抑制作用明显强于BU原药。细胞摄取研究发现BU-NS可增加HepG2细胞对BU的摄取。结论：BU-NS具有增强抑制HepG2细胞增殖与细胞摄取的作用,纳米混悬技术可为蟾毒灵的进一步开发提供新的思路。

Abstract Objective ：To develop bufalin nanosuspensions（BU-NS） and study its characteristics of preparation and effect on proliferation and cellular uptake of HepG2 cells, nethods：BU-NS was prepared by anti-solvent precipitation examined by bufalin （ BU ） was used for method. The trans and quantitative particle size was measured with a particle size analyzer and the morphology was .electron microscopy（TEM）. MTr assay was used to detect the inhibitory effect of on the proliferation of HepG2 cells,high performance liquid chromatography（HPLC） study of cellular uptake. Results：The mean particle size was （390.0± 8.1 ）nm,polydispersion index was 0.22＋0.04,and Zeta potential values was （-9.05 ± 1.21）mV. BU-NS particles was of irregular sphere shape under TEM,and the particle diameter was similar to that determined by particle size analyzer. A short-term stability study showed that BU-NS were physically stable after storage at 4 ℃ for over 15 days. The experiment of cell proliferation inhibition showed that BU-NS inhibited the proliferation of HepG2 cells more strongly than BU within 4 hours. Cellular uptake study showed that BU-NS increased the uptake of BU by HepG2 ceils. Conclusions：BU-NS can enhance inhibition of cell proliferation and cellular uptake of HepG2 cells,and nanosuspension technology can be used as a potential delivery formulation for the further development of BU.