叶酸介导表没食子儿茶素没食子酸白蛋白纳米粒的制备及其体外靶向性与活性评价  

Preparation,activity and targeting ability evaluation in vitro on folate mediated epigallocatechin-3-gallate albumin nanoparticles

作　　者：祖元刚[1] 袁帅[1] 赵修华[1] 张俞[1] 张晓楠[1] 姜茹[1] 

机构地区：[1]东北林业大学森林植物生态学教育部重点实验室,黑龙江哈尔滨150040

出　　处：《药学学报》2009年第5期525-531,共7页Acta Pharmaceutica Sinica

基　　金：国家科技支撑计划资助项目(2006BAD18B0401)

摘　　要：研究能主动靶向于前列腺癌细胞PC-3的表没食子儿茶素没食子酸(EGCG)白蛋白(BSA)纳米粒的制备工艺与体外靶向性和活性评价。去溶剂法制备叶酸介导EGCG白蛋白纳米粒(FA-EGCG-BSANP),采用原子力显微镜(AFM)观察纳米粒形状和粒径,HPLC测定EGCG的载药量和包封率,紫外分光光度法测定叶酸偶联量,以激光共聚焦和荧光分光光度计测定FA-EGCG-BSANP对PC-3细胞的靶向性,用MTT法测定其体外活性。所得FA-EGCG-BSANP的平均粒径为200nm左右,分布均匀;EGCG的包封率可达(81.5±1.8)%,载药量为(29.3±0.6)%,叶酸偶联量为18.363μg.mg-1BSA;PC-3细胞对FA-EGCG-BSANP的摄取量为EGCG-BSANP的23.65倍,并且呈现较强的浓度依赖性;同等浓度下FA-EGCG-BSANP对PC-3细胞的抑制率为82.8%,而EGCG溶液和EGCG-BSANP分别为58.6%和55.1%,并且抑制率与PC-3细胞对这些纳米粒的摄取能力呈正相关。FA-EGCG-BSANP能明显提高EGCG对PC-3细胞的靶向效果,并提高了对细胞的致死作用,从而提高了EGCG作为一种潜在抗癌药物的治疗效果,为进一步探索该纳米粒在体内的靶向性、活性和代谢规律提供了实验基础。To study the preparation, activity and targeting ability evaluation in vitro on epigallocatechin-3- gallate （EGCG） bovine serum albumin nanoparticles targeting to PC-3 cells, the folate mediated EGCG bovine serum albumin nanoparticles （FA-EGCG-BSANP） were prepared by desolvation process. The morphology and particle size of the nanoparticles were determined by atomic force microscope （AFM）. HPLC was used to analyse the entrapment efficiency and drug loading rate of EGCG. The amount of folate conjugation on the BSANP was determined by quantitative ultraviolet （UV） spectrophotometer analysis. The targeting ability to PC-3 was observed using laser scanning confocal microscope （LSCM） and fluorophotometer microscope. And the activity of FA-EGCG-BSANP was mensurated by MTT method. The morphology and particle size distribution of FA-EGCG-BSANP were uniform and even with the mean particle size of 200 nm. The entrapment efficiency and loading rate of EGCG were （81.5 ± 1.8） % and （29.3 ± 0.6） %, respectively, and the amount of folate conjugation was 18.363 μg·mg^-1 BSA. The FA-EGCG-BSANP uptakes by cultured PC-3 cells were 23.65 times the amount of EGCG-BSANP in a concentration dependant manner. The lethality of PC-3 cells treated with FA-EGCG-BSA was 82.8%, while those treated with EGCG and EGCG-BSANP were 58.6% and 55.1%, respectively. And lethality of PC-3 cells was positively correlated with the nanoparticles uptake amount. FA-EGCG-BSANP can significantly promote EGCG to PC-3 cells sites and improve their efficacy, which is considered to an experimental foundation for further research on its activity, targeting ability and metabolism in vivo.

关 键 词：表没食子儿茶素没食子酸 叶酸 纳米粒 前列腺癌细胞 PC-3细胞 靶向性 

分 类 号：R943[医药卫生—药剂学]