College of Pharmacy, Dalian Medical University, Dalian 116044, China.
College of Basic Medical Sciences, Dalian Medical University, Dalian 116044, China.
Life Sci. 2016 Nov 15;165:63-74. doi: 10.1016/j.lfs.2016.09.008. Epub 2016 Sep 15.
Heparin sodium (HS)-loaded polylactic-co-glycolic acid-D-α-tocopheryl polyethylene glycol 1000 succinate (PLGA-TPGS) nanoparticles (HPTNs) were prepared as sustained and targeted delivery carriers and combined with oleanolic acid (OA)-loaded PLGA-TPGS nanoparticles (OPTNs) that had been investigated in our previous work to form a combination therapy system for the treatment of liver cancer.
To inspect cellular uptake and evaluate liver-targeting performance by analysing drug concentrations and cryosections, fluorescent probe coumarin-6 and eosin was used in preparations of HS/eosin-loaded, HS/coumarin-6-loaded, and OA/coumarin-6-loaded PLGA-TPGS nanoparticles. All of these NPs were characterized in terms of size, size distribution, surface charge, drug loading, encapsulation efficiency, and in vitro release profile. The apoptosis of HepG2 cells induced by OPTNs combined with HPTNs was determined by Annexin V-FITC staining and PI labelling.
Transmission electron microscopy indicated that all of the nanoparticles were stably dispersed spheres with diameters ranging from 100 to 200nm. The results demonstrated that fluorescent nanoparticles were efficiently internalized into HepG2 and HCa-F cells, and that they exhibited enhanced liver targeting. The combination of HPTNs and OPTNs resulted in effective cell inhibition in vitro and a remarkable synergistic anticancer effect in vivo. The cell apoptosis results indicated that OPTNs combined with HPTNs could induce HepG2 cell apoptosis and exert synergistic effects. In vivo pharmacodynamics analysis using a solid tumour-bearing mouse model indicated that OPTNs combined with HPTNs could suppress tumour growth by 67.61%.
This research suggests that the combined therapy system of OPTNs and HPTNs could be a new means of hepatoma therapy.
制备肝素钠（HS）负载的聚乳酸-共-羟基乙酸-聚乙二醇 1000 琥珀酸酯（PLGA-TPGS）纳米粒（HPTNs）作为持续和靶向递药载体，并与我们之前研究的载熊果酸（OA）的 PLGA-TPGS 纳米粒（OPTNs）联合，形成用于肝癌治疗的联合治疗体系。
通过分析药物浓度和冷冻切片，使用荧光探针香豆素-6 和伊红，对 HS/伊红负载、HS/香豆素-6 负载和 OA/香豆素-6 负载的 PLGA-TPGS 纳米粒进行细胞摄取和肝靶向性能评价。所有这些 NPs 均在粒径、粒径分布、表面电荷、载药量、包封效率和体外释放特性方面进行了表征。通过 Annexin V-FITC 染色和 PI 标记，测定 OPTNs 联合 HPTNs 诱导 HepG2 细胞凋亡的情况。
透射电子显微镜表明，所有纳米粒均为稳定分散的、粒径 100-200nm 的球体。结果表明，荧光纳米粒能有效地被 HepG2 和 HCa-F 细胞内化，并具有增强的肝靶向性。HPTNs 和 OPTNs 的联合使用在体外具有显著的抑制细胞增殖作用，并在体内具有协同抗癌作用。细胞凋亡结果表明，OPTNs 联合 HPTNs 可诱导 HepG2 细胞凋亡并发挥协同作用。在荷瘤小鼠模型的体内药效学分析中，OPTNs 联合 HPTNs 可使肿瘤生长抑制率达到 67.61%。
本研究表明，OPTNs 和 HPTNs 的联合治疗体系可能是肝癌治疗的一种新方法。
