复方黄黛片有效成分纳米粒的制备及其肝癌治疗研究

目的 探讨复方黄黛片有效成分硫化砷、靛玉红和丹参酮联合装载纳米粒的制备及肝癌HepG2和Huh-7细胞的治疗作用。方法 应用单乳溶媒挥发法，制备聚乳酸-聚羟基乙酸共聚物联合装载硫化砷、靛玉红和丹参酮纳米粒。氢化物发生原子吸收分光光度法和高效液相色谱法分析纳米粒中药物的载药率、包封率和模拟体外释药，利用扫描电镜观察纳米粒的形态，采用粒度分析仪检测纳米粒粒径。荧光显微镜观察HepG2和Huh-7细胞吞噬摄取纳米粒，采用MTT测定法检验细胞活力，细胞存活分析检测细胞克隆形成能力。结果 联合载药纳米粒呈球形，粒径为(115.09 ± 36.71)nm，多分散指数0.131。硫化砷、靛玉红和丹参酮的载药率和包封率分别为(5.17 ± 1.26)%、(1.03 ± 0.45)%、(1.72 ± 0.67)%和(16.15 ± 4.7)%、(76.74 ± 6.8)%、(83.09 ± 9.4)%。硫化砷、靛玉红和丹参酮模拟体外释药曲线早期呈爆发释放，随后为缓慢持续释放。荧光显微镜结果可见肝癌细胞对纳米粒的吞噬摄取，MTT检测显示药物单体和联合载药纳米粒作用后细胞活力较对照降低，联合载药纳米较药物单体粒作用显著，细胞存活分析结果也进一步证实上述结果。结论 复方黄黛片有效成分硫化砷、靛玉红和丹参酮联合装载纳米粒显示出明确的肝癌治疗效果，这为中药的临床给药提供了新的剂型。

The Research on the Therapy on Hepatocellular Carcinoma of Nanoparticles Loading Effective Components of Compound Huangdai Tablets

Objective To investigate the preparation of nanoparticles loading the effective components of Compound Huangdai Tablets(Arsenic sulfide, Indirubin and Tanshinone) and the therapeutic effect of HepG2 and Huh-7 cells. Methods The poly(D, L-lactide-co-glycolide) nanoparticles loading Arsenic sulfide, Indirubin and Tanshinone were prepared by o/w emulsification-solvent evaporation method. The drug loading efficiency, encapsulation efficiency and release profile in vitro were analyzed using Hydride generation atomic absorption spectrometry and High-performance liquid chromatography. The morphology and size of nanoparticles were investigated by scanning electron microscopy and particle size analyzer, respectively. Cell viabilities were measured by MTT assay and forming colonies. Results The nanoparticles were spherical, with a diameter of(115.09 ± 36.71)nm and a polycentrifugation index of 0.131. The drug encapsulation efficiency and loading efficiency of Arsenic sulfide, Indirubin and Tanshinone were(5.17 ± 1.26)%, (1.03 ± 0.45)%, (1.72 ± 0.67)% and (16.15 ± 4.7)%, (76.74 ± 6.8)%, (83.09 ± 9.4)%, respectively. The mimics the drugs release in vitro were biphasic with early bursting release followed by slow and sustained release. Fluorescence microscopy revealed phagocytosis of nanoparticles by HepG2 and Huh-7 cells. MTT detection and cell survival assay showed decreased cell viability after treatment with free drugs and drug loading nanoparticles compared with control cells, and drug loading nanoparticles were more significant than free drugs. Conclusion The nanoparticles loading Arsenic sulfide, Indirubin and Tanshinone have definite therapeutic efficacy on hepatocellular carcinoma, providing a new dose type for the clinical administration of traditional Chinese medicine.