姜黄素纳米微粒的制备及其对人视网膜色素上皮细胞周期的影响

Preparation of curcumin nanoparticles and its effect on human retinal pigment epithelial cell cycle

目的探讨壳聚糖脱氧胆酸负载姜黄素纳米微粒的制备方法,观察姜黄素纳米微粒对人视网膜色素上皮细胞周期的影响。方法以脱氧胆酸基接枝的壳聚糖为载体,姜黄素为负载药物合成姜黄素纳米微粒,测定纳米微粒的负载率与载药量。观察纳米微粒的外形结构,测定其体外释放量。检测不同浓度的壳聚糖脱氧胆酸纳米粒作用于人视网膜色素上皮细胞24、48 h光密度(OD)值,同时观察其细胞形态。测定姜黄素纳米微粒及姜黄素对人视网膜色素上皮细胞周期时相变化。结果姜黄素与壳聚糖脱氧胆酸混合制成负载药物的壳聚糖脱氧胆酸纳米微粒为淡黄色,未负载药物的壳聚糖脱氧胆酸纳米微粒呈类球形或球形,平均粒径为30~50 nm,负载姜黄素的壳聚糖脱氧胆酸纳米微粒粒径为70~100 nm;载药量为27.5%,负载率为55.0%。96 h后药物从纳米微粒中的释放达到平衡,药物的累积释放量达31.6%。不同浓度的壳聚糖脱氧胆酸纳米微粒作用于人视网膜色素上皮细胞24、48 h后OD值与对照比较,差异无统计学意义(P>0.05)。各浓度姜黄素纳米微粒与姜黄素分别对体外培养的人视网膜色素上皮细胞作用24 h后,均出现S期细胞百分比下降,G_(0)~G_(1)期细胞百分比上升。结论通过壳聚糖脱氧胆酸包载的姜黄素纳米微粒能持续释放出姜黄素,缓释功能较好。姜黄素纳米微粒可以阻滞人视网膜色素上皮细胞周期于S期。

Objective To investigate the preparation of curcumin nanoparticles supported by chitosan deoxycholic acid,and to observe the effect of curcumin nanoparticles on human retinal pigment epithelial cell cycle.Methods Curcumin nanoparticles were synthesized by using chitosan grafted with deoxycholate group as carrier and curcumin as supported drug.The load factor and drug-loading rate of the nanoparticles were determined.The shape and structure of the nanoparticles were observed and the amount of their release in vitro was determined.The optical density(OD)values of human retinal pigment epithelial cells treated with chitosan deoxycholic acid nanoparticles at 24 and 48 h were measured,and the cell morphology was observed.The changes of curcumin nanoparticles and curcumin on human retinal pigment epithelial cell cycle were determined.Results Chitosan deoxycholic acid nanoparticles mixed with curcumin and chitosan deoxycholic acid were light yellow.Chitosan deoxycholic acid nanoparticles of unloaded drug were spherical or spherical with an average particle size of 30 to 50 nm,and chitosan deoxycholic acid nanoparticles loaded with curcumin had an average particle size of 70 to 100 nm;the drug-loading rate was 27.5%and the load factor was 55.0%.AfteR96 h,the drug release from the nanoparticles reached equilibrium,and the drug accumulate release rate reached 31.6%.OD values of human retinal pigment epithelial cells treated with different concentrations of chitosan deoxycholic acid nanoparticles foR24 and 48 h were not significantly different from those of the control group(P>0.05).AfteR24 h treatment with curcumin nanoparticles and curcumin,the percentage of cells in S phase decreased and that in G_(0) to G_(1) phases increased.Conclusion Curcumin nanoparticles coated with chitosan deoxycholic acid can continuously release curcumin with good slow-release function.Curcumin nanoparticles can block the human retinal pigment epithelial cell cycle from S phase.