共载光敏剂/UCNPs/双氢青蒿素脂质体的制备及其体外抗肿瘤活性评价  

Preparation of co-loading photosensitizer/UCNPs/dihydroartemisinin liposomes and evaluation of its in vitro antitumor activity

作　　者：彭剑青 邹颖 徐金转 刘丽娜[1] 陶玲[1] 黄静[1] 肖长帅 陈艺 PENG Jian-qing;ZOU Ying;XU Jin-zhuan;LIU Li-na;TAO Ling;HUANG Jing;XIAO Chang-shuai;CHEN Yi(Guizhou Province Specialized Natural Medicine Resource Efficient Utilization Engineering Center(Key Laboratory of Pharmacology and Pharmacological Evaluation of Natural Medicines in Guizhou Higher Education Institutions,Guizhou Medical University-Guiyang Joint Key Laboratory,Key Laboratory of Utilization of Natural Medicine Resources),College of Pharmacy,Guizhou Medical University,Guiyang 550025,China)

机构地区：[1]贵州医科大学,贵州省特色天然药物资源高效利用工程中心(贵州省高等学校天然药物药理与成药性评价特色重点实验室,贵州医科大学-贵阳市联合重点实验室,天然药物资源优效利用重点实验室),药学院,贵州贵阳550025

出　　处：《中草药》2020年第16期4151-4159,共9页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金地区科学基金项目(81960647);贵州省科技厅科学技术基金项目(黔科合基础[2019]1255号);贵州省中医药管理局中医药、民族医药科学技术研究基金(QZYY-2018-093);贵州医科大学博士启动基金(YJ2017-27);国家级大学生创新创业训练计划项目(201810660018)。

摘　　要：目的开发共载光敏剂玫瑰红(rose-bengal,RB)/上转换纳米粒(upconverting nanoparticles,UCNPs)/双氢青蒿素(dihydroartemisinin,DHA)的脂质体(LIP-RUD),并初步考察其体外抗结肠癌活性。方法采用溶剂热法和配体交换法制备水溶性UCNPs,通过薄膜分散法包载RB/UCNPs/DHA得到LIP-RUD;采用HPLC法检测LIP-RUD的包封率,用透射电子显微镜和粒径仪考察其理化性质;通过SOSG探针检测脂质体产生活性氧(ROS)的效率;采用激光共聚焦显微镜观察HCT-116结肠癌细胞对LIP-RUD的摄取情况,并用MTT法评价其体外抑制细胞生长的效果。结果制备得到的LIP-RUD粒径约150 nm,表面电位约-12 mV,RB和DHA包载效率分别达到54.5%和86.5%。UCNPs介导的光敏剂能量转换效率达49.8%;激光照射后,有大量单线态氧(singlet oxygen,1O_2)的生成,LIP-RUD中DHA的12 h累积释放率达到74.9%,相比于无光照组提高了25.6%。细胞实验表明,LIP-RUD能够显著提高药物对结肠癌细胞生长抑制作用,在激光照射下半数有效抑制浓度(IC_(50))为15.33μmol/L。结论 LIP-RUD为治疗结肠癌提供了光动力化疗联用的新思路,该脂质体平台有望增强光动力治疗的体内穿透性和光动力化疗的联合作用效果。Objective To develop the photosensitizer rose-bengal(RB)/upconverting nanoparticles(UCNPs)/dihydroartemisinin(DHA) co-encapsulated liposomes(LIP-RUD) and preliminarily study the in vitro inhibition effects on human colon cancer. Methods The hydrophilic UCNPs were synthesized by solvothermal and ligand conversion and RB/UCNPs/DHA were encapsulated by thin-film dispersion method to obtain LIP-RUD. HPLC was performed to determine the loading ratio(LR) of RB and DHA. Zetasizer was used to evaluate the physiochemical properties of liposomes. The production of ROS was investigated by SOSG probe. In vitro cellular uptake of LIP-RUD was observed by confocal laser scanning microscopy(CLSM) and the cytotoxicity on HCT-116 cells was estimated by MTT assay. Results LIP-RUD showed an average particle diameter of 150 nm with zeta potential of-12 mV. The LR of RB and DHA were 54.5% and 86.5%, respectively. The energy conversion efficiency of UCNPs and RB reached 49.8%. After irradiation, the singlet oxygen(1O2) was generated and 74.9% of encapsulated DHA was released from LIP-RUD at 12 h, which showed an improvement of up to 25.6% compared to the absence of laser irradiation group. In cellular experiments, LIP-RUD exerted improved cytotoxicity on HCT-116 cells. IC50 was 15.33 μmol/L under laser irradiation. Conclusion LIP-RUD provides a new thought in the treatment of human colon cancer by the combination of photodynamic therapy(PDT) and chemotherapy, which is expected to enhance the penetration depth of PDT and the therapeutic effect of combination therapy.

关 键 词：上转换纳米粒 玫瑰红 双氢青蒿素 活性氧 脂质体 光敏剂 结肠癌 溶剂热法 配体交换法 薄膜分散法 细胞摄取 MTT法 单线态氧 抑制作用 

分 类 号：R283.6[医药卫生—中药学]