齐墩果酸纳米粒与肝素钠纳米粒联合应用治疗肝癌的体内外研究
Therapeutic effects of oleanolic acid nanoparticles combined with heparin sodium nanoparticles on liver cancer in vivo and in vitro

摘要：目的 考察自制齐墩果酸纳米粒(OPTN)与自制肝素钠纳米粒(HPTN)联用后对肝癌的体内外抑制作用，并通过对比单独用药的结果，研究两药联用是否具有协同作用。方法 以香豆素6和曙红为标记物，考察人肝癌细胞HepG2和小鼠腹水型肝癌细胞HCa-F对两种纳米粒的摄取情况，利用水溶性四氮唑(WST-1)法和流式细胞仪测定两药合用对HepG2的协同抑制作用以及细胞凋亡情况。体内通过测定肿瘤抑制率和苏木精&曙红染色(H&E)法评估两药合用后的抗肿瘤效果。结果 经过4h的孵育，在HepG2细胞和HCa-F细胞里均能观测到纳米粒发出的荧光，表明包载药物和荧光标记物的纳米粒能被两种细胞摄取；OPTN与HPTN联用能协同抑制肝癌细胞的活性；体内试验结果表明两药联用后能有效抑制肿瘤生长。结论 OPTN与HPTN联用后能产生协同作用，有效抑制肝癌细胞及肝癌实体瘤的生长，作用明显强于单独用药。
ABSTRACT: Objective To investigate the therapeutic effects of oleanolic acid-loaded polylactic-co-glycolic acid-D-α-tocopheryl polyethylene glycol 1000 succinate (PLGA-TPGS) nanoparticles (OPTN) combined with heparin sodium-loaded PLGA-TPGS nanoparticles (HPTN) on liver cancer, and explore whether OPTN combined with HPTN has a synergistic effect by comparing the results of single medication. Methods Coumarin 6 and eosin were used as fluorescent probes to examine the cellular uptake by human liver cancer HepG2 cell and murine ascitic hepatocarcinoma cell strain with high metastasis potential in the lymphatic system (HCa-F). The in vitro cytotoxic combination effect and apoptosis of HepG2 cells induced by OPTN combined with HPTN were also determined using WST-1 assay and Annexin V-FITC staining. The antitumor effect in vivo was determined by tumor growth inhibition rate and hematoxylin & eosin staining (H & E) method. Results Both OPTN with green fluorescence and HPTN with red fluorescence were found in HepG2 cells and HCa-F cells, suggesting that they had been internalized. The cell cytotoxicity test and Annexin V-FITC staining results showed that OPTN combined with HPTN had a synergistic effect. The therapeutic effect in vivo showed that OPTN combined with HPTN effectively inhibited tumor growth better than the drug alone. Conclusion OPTN combined with HPTN has a synergistic effect in more effectively inhibiting liver cancer better than single medication