靶向5-氟尿嘧啶纳米微球对体外培养的人晶状体上皮细胞作用的研究  

Experimental study on 5-fluourouracil-loaded immunonanoparticles targeting to lens epithelial cells

作　　者：李筱荣[1] 刘新玲 常津[2] 刘小燕[2] 

机构地区：[1]天津医科大学眼科中心,300070 [2]天津大学材料学院纳米生物研究所

出　　处：《中华眼科杂志》2006年第6期526-530,共5页Chinese Journal of Ophthalmology

基　　金：天津市自然科学基金资助项目(003702711)

摘　　要：目的评价靶向人晶状体上皮细胞免疫载药纳米微球HILE6-PLA(5-FU)-NP的免疫学特征及体外对靶细胞的特异性结合和杀伤作用。方法采用碳二亚胺法将5-氟尿嘧啶(5-FU)聚乳酸纳米微球PLA(5-FU)-NP与抗人晶状体上皮细胞单克隆抗体HILE6连接,制备免疫载药纳米微球。测定其中5-FU与HILE6物质的量之比;ELISA法检测连接后单克隆抗体HILE6的活性变化。设免疫纳米微球组、HILE6与载药纳米微球混合组、载药纳米微球组、空白对照组,MTT法检测对晶状体上皮细胞增殖的抑制作用。间接免疫荧光法观察单克隆抗体HILE6、免疫纳米微球和载药纳米微球对兔晶状体上皮细胞的附着、内化过程。结果免疫纳米微球中5-FU与HILE6物质的量之比约为1809∶1,单克隆抗体HILE6保留免疫活性可达原抗体的84%。免疫纳米微球对晶状体上皮细胞作用2h的半数抑制剂量IC50为5.0μg/ml,抑制作用较载药纳米微球组、单克隆抗体与微球混合组有明显提高。免疫纳米微球在30min内可识别并附着于兔晶状体上皮细胞;2h进入细胞质均匀分布;4h后集中于细胞核区。结论免疫纳米微球HILE6-PLA(5-FU)-NP可携带大量药物,保持特异的免疫活性,在体外识别并进入晶状体上皮细胞有效抑制其增殖。Objective To evaluate the immunological characteristics of an immunonanoparticles targeting to human lens epithelial cells and to study if it could internalize into and inhibit the proliferation of target cells in vitro. Methods Crosslinker carbodiimide was used to couple McAb HILE6 （anti-lens epithelial cells antibody） with 5-fluourouracil-loaded polyactic acid nanoparticles PLA（5-FU）-NP to prepare the immunonanoparticles HILE6-PLA （5-FU）-NP. The molar ratio of HILE6 and 5-FU in the immunonanoparticles were observed. The immunological activity of the antibodies in the immunonanoparticles was assessed by ELISA. The third passage lens epithelial cells were divided into four groups; immunonanoparticles, 5-FU nanoparticles, mixed HILE6 and 5-FU nanoparticles and the controls. The proliferation of lens epithelial cells were evaluated by MTT analysis. Internalization of immunonanoparticles, 5-FU nanoparticles and McAb HILE6 were observed by indirect immunofluorescence study of lens epithelial cells. Results The molar ratio of 5-FU to HILE6 in the immunonanoparticles was 1809：1 and 84% of the original immunological activity of antibodies could be retained. The immunonanoparticles inhibited the proliferation of lens epithelial cells, which was significantly greater than original 5-FU nanoparticles or the blend group. The IC50 inhibition of immunonanoparticles was 5.0 μ/ml 2 hours after treatment. The immunonanoparticles were bound to the cells surface in 30 minutes; internalized in 2 hours and accumulated around the nucleus after 4 hours. Conclusion The immunonanoparticles retain immunological activity and could specifically internalize into the lens epithelial cells to inhibit their proliferation. （Chin J Ophthalmol, 2006,42 ： 526-530）

关 键 词：免疫纳米微球 靶向治疗 抗体 单克隆 晶状体上皮细胞 

分 类 号：R776.1[医药卫生—眼科] R979.1[医药卫生—临床医学]