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（polyEGCG），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pH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polyEGCG和Mel在B16F10和A549细胞的协同抗肿瘤效应。结果 合成的polyEGCG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/mL polyEGCG溶液逐滴加入到1 mg/mL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，polyEGCG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 (polyEGCG) 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 pH. The cell uptake of FITC-labeled Mel and EMN was measured by flow cytometry. The synergistic antitumor effect of polyEGCG and Mel was investigated using B16F10 and A549 cells by MTT assay. Results The structure of polyEGCG was confirmed by ¹H-NMR and MS. The optimized preparation process of EMN was as follws:0.5 mg/mL polyEGCG solution was added dropwise to 1 mg/mL 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 polyEGCG and Mel was less than 1, which intimated the synergistic effect when combined therapy with polyEGCG 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.

国家自然科学基金资助项目（81503259）；江苏省自然科学基金资助项目（BK20151002）；江苏高校优势学科建设工程资助项目（PPZY2015A070）；江苏省研究生创新工程项目；南京中医药大学大学生创新创业项目