负载5-ALA纳米微球对膀胱癌光动力学作用的初步研究

Effect of 5-ALA derivative-loaded photosensitized nanoparticles photodynamic therapy on bladder cancer T24 cells

目的探讨负载5-盐酸乙酰氨基丙酸(5-aminolevulinic acid,5-ALA)的光敏感纳米微球的制备方法,并初步研究其在体外对膀胱癌T24细胞的光动力学效应。方法以纳米沉淀法制备光敏感纳米微球,紫外分光光度法检测载药率,动态光散射的方法检测纳米粒径,原子力显微镜观察纳米微球形貌。将膀胱癌T24细胞分别与不同浓度的光敏感纳米微球共同孵育,以能量密度为6 J/cm2,波长为650 nm半导体激光照射。另设空白对照组、空载体组、5-ALA组。以MTT比色法测定T24细胞的生长抑制率。结果光敏感纳米微球载药量为7%,负载效率85%。处理组5、10、25和50μg/ml光敏感纳米微球的细胞生长抑制率分别为73.19%、79.95%、83.86%和89.74%,与空白对照组、空载体组、5-ALA组比较差异有统计学意义(P<0.05)。结论纳米微球负载5-ALA具有理想的载药率,光敏感纳米微球能显著提高光动力学杀伤效应。

Objective To develop a novel 5-ALA （5-aminolevulinic acid） derivative-loaded photosensitized nanoparticles, and to study the effect of photodynamic therapy on bladder cancer T24 cells. Methods Drug-loaded photosensitized nanoparticles were prepared by nanoprecipitation method, using the UV-spectrophotometric method to determine the drug loading content. Mean diameter and size distribution of the prepared nanoparticles were determined by dynamic light scattering. Atomic force microscope was used to study the surface morphology of nanoparticles in a greater detail. At test groups, T24 cells were incubated with different concentrations of photosensitized nanopartieles, and then irradiated by 650nm laser （ 6J/cm^2 ）. Additionaly, as control group, there were following ones blank group, nanoparticles and irradiation group, 5-ALA and irradiation group. MTT method was used to detect the growth inhibitory rate of T24 cells. Results The yield of sample was 85% and the drug loading content was 7%. The growth inhibitory rates of 5.0μg/ml, 10. 0μg/ ml, 25. 0μg/ml, 50. 0μg/trd photosensitized nanoparticles were 73.19% ,79. 95% ,83. 86% and 89. 74%, respectively. Compared with them, the difference was statistically significant （P 〈 0. 05 ）. Conclusion The drug loading content of 5-ALA derivative-loaded photosensitized nanoparticles was satisfied, and can significantly improve the effect of photodynamic therapy on T24 cells.