作　　者： (方栋）; (张蕾）; (孙娟）; (吴文瀚）; (乔宏志）; (狄留庆）;

机构地区： 南京中医药大学药学院,江苏南京210023 江苏省中药高效给药系统工程技术研究中心,江苏南京210023 南京市中药微丸产业化工程技术研究中心,江苏南京210023

出　　处： 《中草药》 2017年第16期3300-3307,共8页

摘　　要： 目的制备具有协同抗肿瘤作用的茶多酚-蜂毒肽纳米复合物(EMN),并考察其抗肿瘤活性。方法通过拜耳反应合成了多聚表没食子儿茶素没食子酸酯(poly EGCG),~1H-NMR和MS表征其结构和相对分子质量。通过正交试验优化EMN的制备工艺,并考察在不同p H值条件下蜂毒肽(Mel)的体外释放。通过流式细胞仪比较异硫氰酸荧光素(FITC)标记的Mel和EMN的细胞摄取情况。MTT法考察poly EGCG和Mel在B16F10和A549细胞的协同抗肿瘤效应。结果合成的poly EGCG用~1H-NMR和MS确证了结构,正交试验优化的EMN最佳制备工艺为将0.5 mg/m L poly EGCG溶液逐滴加入到1 mg/m L等体积的Mel溶液中,边滴加边搅拌,室温孵育30 min,即得。体外释放实验表明EMN释放具有酸响应性。摄取实验表明Mel和EMN均能被肿瘤细胞摄取,摄取量相当。MTT实验结果表明,poly EGCG和Mel联合使用的协同系数(CI)小于1.0,具有协同抗肿瘤效果。结论采用自组装的方式制备EMN,工艺简单,粒径适宜,且具有协同抗肿瘤的效果,值得深入研究。 Objective To prepare nanocomplex comprising tea polyphenol and melittin（EMN） with synergistic effect for antitumor therapy. Methods Poly-epigallocatechin gallate（poly EGCG） was synthesized by Bayer reaction and characterized by -1H-NMR and MS spectra. The preparation process of EMN was optimized by orthogonal test, and the in vitro release of EMN was conducted at different medium with different p H. The cell uptake of FITC-labeled Mel and EMN was measured by flow cytometry. The synergistic antitumor effect of poly EGCG and Mel was investigated using B16F10 and A549 cells by MTT assay. Results The structure of poly EGCG was confirmed by -1H-NMR and MS. The optimized preparation process of EMN was as follws： 0.5 mg/m L poly EGCG solution was added dropwise to 1 mg/m L equal volume of Mel solution, and then incubated at room temperature for 30 min. In vitro release studies showed that acidic environment could accelerate the release of Mel. Cell uptake experiments showed that the cell uptake of Mel and EMN was comparable. MTT assay results showed the combination index（CI） of poly EGCG and Mel was less than 1, which intimated the synergistic effect when combined therapy with poly EGCG and Mel. Conclusion EMN was prepared by a self-assembly method, which has a simple preparation process, a suitable particle size, and synergistic antitumor effect, and it is worthy of further studies.

关 键 词： 药辅合一 蜂毒肽 茶多酚 纳米复合物 表没食子儿茶素没食子酸酯 体外释放 细胞摄取 抗肿瘤 协同效应 正交试验 法 自组装