麦胚凝集素修饰的 EGCG-明胶-壳聚糖纳米粒的制备、表征及体外抗肿瘤活性研究

摘要

OBJECTIVE To preparation wheat germ agglutinin (WGA)-conjugated EGCG-gelatin-chitosan nanoparticles, and evaluate its anti-tumor activity in vitro.METHODS EGCG-Gel-Cs nanoparticles were fabricated based on an electrostatic bonding between gelatin and chitosan.The formulation variables were optimized by Box-Behnken Design (BBD) of response surface methodology(RSM) of encapsulation efficiency(Y ,%) as dependent variable.The surface of nanoparticles was then modified by glutaraldehyde-activated WGA to obtain WGA-EGCG-Gel-Cs nanoparticles.The differential scanning calorimetry was used to analyze its material phase of the drug in nanoparticles.Cell toxicity and apoptosis were employed to evaluate the anti-tumor activities.RESULTS The optimal formula was as follows:the mass ratio of Gel and Cs was 4.2,Gel and EGCG was 2.82 and the temperature was 34 ℃.The encapsulation efficiency of EGCG-Gel-Cs nanoparticles was (74.42 ±0.074)%with the particle size(264.13±6.48)nm.After modifying,the particle size increased to(389.70±9.00)nm.Simultaneous-ly,cytotoxicity and apoptosis of WGA-EGCG-Gel-Cs nanoparticles was significantly higher than that of free EGCG.CONCLU-SION Prepared WGA-EGCG-Gel-Cs nanoparticles can significantly enhance tumor cell toxicity and apoptosis of EGCG.%目的：制备麦胚凝集素(WGA)修饰的表没食子儿茶素没食子酸酯(EGCG)-明胶(Gel)-壳聚糖(Cs)纳米粒,研究其对结肠癌 HT-29细胞的作用。方法以明胶及壳聚糖为载体辅料,采用静电自组装的方法制备 EGCG-Gel-Cs 纳米粒,以包封率为指标,通过 Box-Behnken 设计-效应面法(BBD-RSM)优化纳米粒的制备处方及工艺,再将经戊二醛活化的 WGA 修饰到纳米粒表面。采用差示扫描量热分析药物在纳米粒中的存在状态,采用细胞毒性及细胞凋亡实验,比较研究纳米粒与原料药的体外抗肿瘤活性。结果 EGCG-Gel-Cs 纳米粒的最优处方：Gel 与 Cs 的质量比为4.2,Gel 与 EGCG 的质量比为2.82,反应温度为34℃,包封率为(74.42±0.074)%。EGCG-Gel-Cs 纳米粒粒径为(264.13±6.48)nm,经过修饰后纳米粒粒径为(389.70±9.00)nm。WGA-EGCG-Gel-Cs 纳米粒对 HT-29细胞的细胞毒性和细胞凋亡率显著高于 EGCG 原料药。结论 WGA-EGCG-Gel-Cs 纳米粒可显著提高细胞毒性作用。