巨噬细胞运载叶酸靶向携氧微泡造影剂的初步探讨  

Study of macrophages as cell carriers to deliver fioate modified oxygen loaded contrast agent

作　　者：何娟[1] 常淑芳[1] 孙江川[1] 朱深银[2] 魏娜[1] 骆婷婷[1] 马容[1] 王志刚[3] 

机构地区：[1]重庆医科大学附属第二医院妇产科,400010 [2]重庆医科大学附属第一医院药学部 [3]重庆医科大学超声影像学研究所

出　　处：《中华超声影像学杂志》2016年第2期178-182,共5页Chinese Journal of Ultrasonography

基　　金：国家自然科学基金(81372799，81572558)@@@@National Natural Science Foundation of China

摘　　要：目的探讨巨噬细胞作为叶酸靶向多模态超声微泡影剂载体的可行性。方法采用墨汁吞噬实验及免疫荧光法分别检测体外培养的巨噬细胞吞噬活性和细胞膜表面叶酸受体（FRs）表达情况。采用机械震荡法合成普通携氧荧光脂质微泡造影剂（OPLMB）及叶酸靶向携氧荧光脂质微泡造影剂（TOPLMB），与体外培养的巨噬细胞进行共孵育。根据处理因素不同将细胞分为3组：A组，OPLMB组；B组，TOPLMB＋游离叶酸阻断组；C组，TOPLMB组。采用荧光显微镜观察细胞内荧光强度，普通光镜观察巨噬细胞对各组微泡的吞噬率。结果体外培养的巨噬细胞对墨汁的吞噬率为（99．3±1．0）％，免疫荧光显示其表面高表达FRs。巨噬细胞与微泡体外共孵育30min，荧光显微镜示C组细胞内荧光强度明显高于A、B组，A、B、C各组吞噬率分别为（19．5±0．2）％、（21．0±0．2）％、（81．2±10．0）％，C组吞噬率明显高于A组及B组（P〈0．05）。结论体外分离培养的巨噬细胞具备高吞噬活性，且该巨噬细胞表面高表达活性FRs，可作为叶酸靶向多模态OPLMB载体。Objective To investigate the feasibility of macrophages as cell carriers to deliver floate modified oxygen loaded ultrasound contrast agent. Methods The phagocytic activity of macrophages was analyzed by ink phagocytose test, and the expression of folate recepters （FRs） on macrophages cell membrane surface was tested by immunofluofluorescence assay. Oxygen/paclitaxel loaded lipid microbubbles（OPLMB） and folate-targeted OPLMB（TOPLMB） were synthesized by mechanical shock method and incubated with macrophages in vitro. According to different treatment conditions, the cells were divided into three groups： group A （OPLMB）, group B （free folic acid ＋ TOPLMB） and group C （TOPLMB）, the fluorescence intensity of the cells were observed under fluorescence microscope, and the phagocytic percentage and the phagocytic index of macrophages uptake OPLMB and TOPLMB were observed by bright field microscope. Results The phagocytic percentage of macrophages phagocytose ink was （99.3±1.0）%, FRs was highly expressed on macropbages cultured in vitro. After incubation for 30 minutes, the fluorescence intensity of group C was significantly higher than those of A and B, the phagocytic percentage in three groups were （19.5 ±0.2）%,（21.0±0.2）M and （81.2±10.0）% respetively. The phagocytic percentage of group C were significantly higher than those in group A and group B （ P 〈0.05）. Conclusions Macrophages cultured in vitro possess highly phagocytic activity and these cells highly express FRs,and can be used as cell carriers to deliver floate modified oxygen loaded multimodality ultrasound contrast agent.

关 键 词：微气泡 巨噬细胞 叶酸受体 细胞吞噬作用 

分 类 号：R943[医药卫生—药剂学]