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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 investigate the preparation process and in vitro targeting and activity of epigallocatechin gallate (EGCG) albumin (BSA) nanoparticles that can actively target prostate cancer cells PC-3. Folate-mediated EGCG albumin nanoparticle (FA-EGCG-BSANP) was prepared by solvent removal method. The shape and size of nanoparticles were observed by atomic force microscopy (AFM). The drug loading and entrapment efficiency of EGCG were determined by HPLC. Ultraviolet spectrophotometry was used. The coupling amount of folic acid was measured, and the targeting ability of FA-EGCG-BSANP to PC-3 cells was determined by laser confocal and fluorescence spectrophotometry. The in vitro activity of FA-EGCG-BSANP was determined by MTT assay. The average particle size of the obtained FA-EGCG-BSANP was about 200 nm with a uniform distribution; the encapsulation efficiency of EGCG was (81.5±1.8)%, the drug loading was (29.3±0.6)%, and the coupling amount of folic acid was 18.363 μg. The uptake of FA-EGCG-BSANP by PC-3 cells was 23.65-fold higher than that of EGCG-BSANP, and showed a strong concentration-dependent manner; the inhibitory rate of FA-EGCG-BSANP on PC-3 cells at the same concentration. It was 82.8%, while EGCG solution and EGCG-BSANP were 58.6% and 55.1%, respectively, and the inhibition rate was positively correlated with PC-3 cells uptake of these nanoparticles. FA-EGCG-BSANP can significantly increase the targeting effect of EGCG on PC-3 cells and increase the lethal effect on cells, thereby improving the therapeutic effect of EGCG as a potential anti-cancer drug, and further exploring the nanoparticles. The in vivo targeting, activity and metabolism provide the experimental basis.