叶酸靶向荧光量子点脂质体纳米探针的体内外评价研究  

Biological Application of Folate-Targeted Fluorescent Quantum Dots Liposome Nanoprobes

作　　者：杨畅 胡珺 赵青 陈思颖 刘亭[1,2] 

机构地区：[1]贵州医科大学药学院,贵阳550025 [2]贵州省药物制剂重点实验室,贵阳550004 [3]国家苗药工程技术研究中心,贵阳550004 [4]民族药与中药开发应用教育部工程研究中心,贵阳550004

出　　处：《中国药学杂志》2018年第3期210-217,共8页Chinese Pharmaceutical Journal

基　　金：贵州省科学技术基金项目[黔科合字(2014)2019号];贵州省卫生计生委科学技术基金项目(gzwjkj2015-1-027);贵州省科技厅-贵州医科大学科学技术联合基金项目[黔科合LH字(2016)7364号];国家自然科学基金项目(81603189);贵州省科技厅团队项目[黔科合平台人才(2016)5613\5677]

摘　　要：目的对自制的叶酸靶向荧光量子点脂质体纳米探针(Folate-QDs-Liposome)进行体内外评价研究,如细胞毒性、肿瘤靶向性、动物毒性等。方法通过细胞增殖抑制实验(MTT)及流式细胞术考察Folate-QDs-Liposome对细胞生长抑制作用和细胞凋亡情况;同时通过荧光染色法对Folate-QDs-Liposome对叶酸受体表达阳性肿瘤细胞的靶向性进行确认;最后以表皮浸润给药方式考察其在斑马鱼体内的毒性及分布情况。结果 Folate-QDs-Liposome与量子点(QDs)和量子点脂质体(QDs-Liposome)相比较,在同一浓度下QDs细胞毒性最强,Folate-QDs-Liposome、QDs-Liposome两者无明显细胞毒性差异,说明QDs经脂质体包裹后其毒性明显降低。Folate-QDs-Liposome对Hela细胞具有靶向性,但对A549细胞几乎无靶向性;且游离的叶酸可阻断叶酸与肿瘤细胞上叶酸受体的特异性结合,证明Folate-QDs-Liposome通过叶酸-叶酸受体途径进入叶酸表达阳性的肿瘤细胞。在斑马鱼体内从头部开始分布,最后通过尾部排泄出体外。结论 Folate-QDs-Liposome具有潜在的生物医学应用前景,可以在肿瘤早期的检测、诊断中发挥重要作用。OBJECTIVE To illustrate and evaluate the properties of folate-targeted liposomes loaded with quantum dots （folate- QDs-liposomes） nanoprobes in vitro and in vivo, such as cytotoxicity, the targeting of folate-QDs-liposomes nanoprobes for tumor cells mediated by the folate-folate receptor pathway, their toxicity in vivo and so on. METHODS Firstly, the inhibitory effects of folate- QDs-liposomes on cell growth and apoptosis were investigated by MTT and flow cytometry. Secondly, the targeting of folate-QDs-lipo- somes for folate receptor-positive tumor cells was verified by fluorescence staining. Finally, the epidermal infiltration method was adopt- ed to examine the toxicity and distribution in vivo. RESULTS Under the same concentration, folate-QDs-liposomes and QDs-lipo- somes had similar cytotoxicity, and QDs had the most obvious cytotoxicity, which verified that the cytotoxicity of QDs was significantly reduced after liposome coating. In the fluorescence experiment, it was observed that folate-QDs-liposomes targeted at Hela ceils, but did not target at A549 cells. Free folic acid could block the specific binding of folate-QDs-liposomes to folate receptors on tumor cells. It was proved that folate-QDs-liposomes could enter tumor ceils positively expressing folate receptors through the path of folic acid and folate receptors. In vivo, folate-QDs-liposomes nanoprobes could spread throughout the zebrafish body through the blood circulation sys- tem. They were distributed from head and excreted from the tails. CONCLUSION Folate-QDs-liposomes have potential biomedical application prospects, and can be expected to play an important role in the early detection and diagnosis of tumors.

关 键 词：叶酸靶向 荧光量子点 脂质体 毒性 靶向性 体内分布 

分 类 号：R944[医药卫生—药剂学]