荧光标记的叶酸修饰壳聚糖纳米载体研制 - 中南大学学报.pdf

第 41 卷第 1 期 中南大学学报( 自然科学版) Vol.41 No.1 2010 年 2 月 Journal of Central South University (Science and Technology) Feb. 2010 荧光标记的叶酸修饰壳聚糖纳米载体研制 金鑫，张阳德，王吉伟，张丽华，杨璞, 张旭，胡玉，于丽 ( 中南大学 卫生部肝胆肠外科研究中心，湖南 长沙 410008) 摘 要：制备荧光标记的叶酸偶联壳聚糖纳米粒，为抗肿瘤药物给药系统提供载体材料。通过叶酸活性酯与壳聚 糖上的氨基反应，使叶酸与壳聚糖偶联。将异硫氰基荧光素与叶酸偶联壳聚糖进行化学嫁接，以离子交联法制成 具有荧光的叶酸偶联壳聚糖纳米粒，并与肝癌 HepG2 细胞进行体外细胞实验。实验结果表明：叶酸活性酯用量 和反应温度及试剂滴加速度是影响偶联比的主要因素；在叶酸活性酯与壳聚糖用量质量比为 1׃1，反应温度为 30 ℃，滴加速度为2 mL/min ，反应时间为 12 h 的条件下可得到偶联稳定的叶酸偶联壳聚糖；所制得的纳米粒粒径 为 290 nm ，形态规则，细胞荧光效果明显；此方法能用于制备荧光标记的叶酸修饰壳聚糖纳米粒载体。 关键词：叶酸；壳聚糖；纳米粒；异硫氰基荧光素 中图分类号：R979.1; TQ460.6 文献标志码：A 文章编号：1672−7207(2010)01−0161−05 Preparation and characterization of fluorimetric folate-coupled chitosan nano-carrier JIN Xin, ZHANG Yang-de, WANG Ji-wei, ZHANG Li-hua, YANG Pu, ZHANG Xu, HU Yu, YU Li (National Hepatobiliary and Enteric Surgery Research Center, Central South University, Changsha 410008, China) Abstract: Folate-coupled chitosan was prepared as a delivery system material for anticancer drug by the reaction of the activated folate ester with the amine group on the chitosan. The compound of folate-coupled chitosan grafted on fluorescein isothiocyanate (FITC) by chemical bond was prepared. Then FITC-labelled nanoparticles were prepared by ionically cross-linking method, and reaction with HepG2 and cells in vitro experiments. The results show that the amount of the activated folate ester, the speed of adding agent and the reaction temperature are the main factors affecting the coupling ratio. Folate-coupled chitosan is stably coupled when the ratio of the mass of the active folate ester to chitosan