姜黄素-PLGA纳米粒的制备及对人前列腺癌PC-3细胞的体外研究
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国家自然科学基金(30860284);贵阳市科技攻关(2009筑科农合同字第3-009号);贵州省教育厅重点项目(黔教科2007027)


Preparation of curcumin-loaded PLGA nanoparticles and its effects on human prostate cancer PC-3 cells in vitro
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    摘要:

    目的:制备姜黄素(curcumin,Cur)聚乳酸-羟基乙酸(poly lactic-co-glycolic acid,PLGA)纳米粒(Cur-PLGA-NPs),并考察其理化性质-体外释药特性及对PC-3细胞的体外影响。方法:运用乳化-溶剂挥发法制备Cur-PLGA-NPs,利用透射电镜观察纳米粒的外观形态;动态激光粒度仪分析其粒径大小及分布;超速离心法测定载药率和包封率;透析法观测体外释放效果;采用CCK-8比色法检测其对前列腺癌PC-3细胞的增殖抑制情况,流式细胞仪(FCM)进行细胞凋亡率检测,透射电镜观察细胞超微结构,并与单纯Cur进行比较。结果:透射电镜下姜黄素纳米粒呈圆形或椭圆形,平均粒径为(165.36 ± 24.21)nm,包封率为(83.05 ± 1.07)%,载药率为(10.87 ± 0.58)%,体外药物释放试验显示,Cur-PLGA-NPs在最初的24 h 内释放率超过44.02%,10 d累计释放率达84.81%。 5~40 μmol/L Cur及Cur-PLGA-NPs作用PC-3细胞24~72 h后细胞的抑制率(9.38%~83.62% vs. 10.56%~89.53%)。浓度为20 μmol/L和40 μmol/L的Cur和Cur-PLGA-NPs组间比较,差异有统计学意义(P < 0.05)。FCM显示同浓度实验组细胞凋亡率与对照组比较,差异具有统计学意义(P < 0.05)。透射电镜显示Cur-PLGA-NPs作用PC-3细胞后出现典型的细胞凋亡形态学特征。结论:Cur-PLGA-NPs具有良好的药物缓释特性,增强了Cur对PC-3细胞的体外杀伤和抑制增殖能力。

    Abstract:

    Objective:To prepare curcumin-loaded poly lactic-co-glycolic acid nanoparticles(Cur-PLGA-NPs),to characterize their physicochemical properties,and to study the in vitro release behavior and antitumor activity on human prostate cancer PC-3 cells in vitro. Methods:The Cur-PLGA-NPs were prepared by emulsification-solvent evaporation method. The transmission electron microscope was used to observe the particle appearance,Zetasizer instrument was used to detect the diameter,and ultracentrifugation was utilized to determine drug-loading rate and entrapment rate. Dynamic dialysis method was used to study the in vitro release behavior of Cur-PLGA-NPs. The antitumor activity on PC-3 cells was determined by CCK-8 method. Cells apoptosis and apoptosis rate were examined by using flow cytometry. Ultrastructural changes of cells were observed under the electronic microscopy,and compared to free Cur groups. Results:The optimal NPs were round with the nanometric size of(165.36 ± 24.21)nm,high entrapment rate of(83.05 ± 1.07)% and drug-loading rate of(10.87 ± 0.58)%. In vitro release test showed that release rate within 24 h of Cur-PLGA-NPs was 44.02%,and accumulating release rate was 84.81% at day10. After an effection of 5~40 μmol/L Cur and Cur-PLGA-NPs on PC-3 cells for 24~72 h,the inhibition rates of Cur and Cur-PLGA-NPs were 9.38%~83.62% and 10.56%~89.53%. CCK-8 experience revealed that there was significant difference between 20 μmol/L group and 40 μmol/L group compared Cur with Cur-PLGA-NPs(P < 0.05). Flow cytometry indicated that there was a significant difference between experiment group and control group(P < 0.05). Transmission electron microscopy showed that Cur-PLGA-NPs induced morphological change of apoptosis on PC-3 cells. Conclusion:The Cur-PLGA-NPs has good drug sustained-release capacity in vitro,and significantly improves the capacity of Cur to kill and inhibit the proliferation of PC-3 cells in vitro.

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钟思文,姜锡男,陈方敏,石家齐,李登宝,陈绪龙,唐 帅,陈 程.姜黄素-PLGA纳米粒的制备及对人前列腺癌PC-3细胞的体外研究[J].南京医科大学学报(自然科学版),2017,(2):194-198

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  • 收稿日期:2016-03-13
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  • 在线发布日期: 2017-02-27
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