包載姜黃素納米膠束的制備與體外抗腫瘤評價

范子梁，金冰慧，徐霞芳，蔣巧穎，徐荷林

（溫州醫(yī)科大學(xué) 藥學(xué)院，浙江 溫州 325035）

·論 著·

包載姜黃素納米膠束的制備與體外抗腫瘤評價

范子梁，金冰慧，徐霞芳，蔣巧穎，徐荷林

（溫州醫(yī)科大學(xué) 藥學(xué)院，浙江 溫州 325035）

目的：構(gòu)建載姜黃素納米膠束，體外評價其對C6腦膠質(zhì)瘤細(xì)胞的生長抑制作用。方法：合成新型十一烯酸-接枝-ε-多聚賴氨酸（ε-PLL-UNA）聚合物，并對其結(jié)構(gòu)進(jìn)行表征，采用芘熒光探針法對其臨界膠束濃度進(jìn)行測定；以ε-PLL-UNA為材料，采用透析法制備載姜黃素納米膠束，以動態(tài)光散射和透射電鏡對其粒徑、形態(tài)進(jìn)行表征，以動態(tài)透析法測定藥物釋放行為，以激光共聚焦顯微鏡觀察體外細(xì)胞攝取性，以腫瘤球模型考察其體外抗腫瘤作用。結(jié)果：1H-NMR和FT-IR結(jié)果表明ε-PLL-UNA聚合物成功合成，其臨界膠束濃度為0.19 g/L，能自發(fā)組裝成膠束；載姜黃素納米膠束載藥量能達(dá)到12.22%±2.13%、包封率則高達(dá)85.12%±3.64%，平均粒徑為（60.6±2.1）nm、Zeta電位為（28.2±5.6）mV，具有球形微觀結(jié)構(gòu)；該納米膠束48 h釋放84%的姜黃素，體外快速被C6細(xì)胞攝取；與姜黃素溶液相比，該納米膠束能明顯抑制膠質(zhì)瘤細(xì)胞球的生長。結(jié)論：ε-PLL-UNA聚合物膠束對姜黃素具有較高的載藥量，粒徑小于100 nm、分布均勻，體外緩慢釋放藥物，提高了C6細(xì)胞對姜黃素的攝取，而且對C6細(xì)胞球具有有效的殺傷作用。

姜黃素；膠束；聚賴氨酸；十一烯酸；神經(jīng)膠質(zhì)瘤；細(xì)胞球

Abstract: Objective:To prepare a novel polymer micelles encapsulating curcumin (CUR) and evaluate its anti-tumor effect on glioma in vitro.Methods:A novel polymer, undecenoic acid-grafted-ε-polylysine (ε-PLLUNA), was synthesized, and its chemical structure was confirmed by1H-NMR and FT-IR. The CAC value of ε-PLL-UNA polymer was also detected by pyrene fluorescence probe. CUR-loaded micelle was prepared by dialysis method using ε-PLL-UNA as materials, and its particle size and morphology were also studied under dynamic light scattering (DLS) and transmission electron microscope (TEM), respectively. Furthermore, in vitro drug release profiles from CUR-Micelles were explored by dynamic dialysis method. Finally, the cellular toxicity against C6 cells spheroids and the cellular uptake of CUR-Micelles were evaluated.Results:ε-PLL-UNA polymer was successfully synthesized and able to self-assemble into micelles above its CAC value of 0.19 g/L. CUR-loaded micelle had a mean diameter of (60.6±2.1)nm, and zeta potential of (28.2±5.6)mV, exhibiting the spherical shape determined by TEM. Drug loading content and drug loading efficiency for CUR-loaded micelle were high up to 12.22%±2.13% and 85.12%±3.64%, respectively. About 84% of CUR were released from the micelles in 48 hours. CUR-loaded micelle can promote the cellular uptake of its encapsulated CUR by C6 cells, displaying a significantly higher cytotoxicity against C6 cells. Besides, the growth of C6 cells spheroids was significantly inhibited by CUR-loaded micelle.Conclusion:CUR is efficiently encapsulated in ε-PLL-UNA micelles with a particle size of less than 100 nm, which improved the cellular uptake of C6 cells. The sustained-release of CUR from CUR-loaded micelle is also observed. More importantly, CUR-Micelles has superior growth inhibition against C6 cells spheroids.

Key words:curcumin; micelles; polylysine; undecylenic acid; glioma; cells spheroids

姜黃素（curcumin，CUR）是一種天然酚類色素，廣泛存在于姜黃屬植物姜黃、莪術(shù)、郁金等的根莖中，是姜黃的重要活性成分。姜黃素對機(jī)體各系統(tǒng)作用廣泛，具有較好的抗氧化、抗炎、抗動脈粥樣硬化、抗腫瘤等藥理作用，且毒性低、不良反應(yīng)小[1]。……