新型姜黄素纳米胶束滴眼液的制备及体内外性质研究  

Studies on a new nanomicelle curcumin ophthalmic solution:preparation and characterizations in vitro/in vivo

作　　者：李梦双[1,2] 林桂明[1,2] 李君[2] 吴祥根[1,2] 

机构地区：[1]济南大学山东省医学科学院生命与科学学院,山东省济南市250022 [2]山东省眼科研究所,山东省青岛市266071

出　　处：《眼科新进展》2016年第10期901-905,共5页Recent Advances in Ophthalmology

基　　金：国家自然科学基金资助(编号:81271036;81500751);2015年度山东省重点研发计划资助[编号:2015GGC03010(2015GSF-118121)]~~

摘　　要：目的制备新型姜黄素纳米胶束滴眼液,检测其理化性质,并观察其生物学性质。方法通过溶剂蒸发-水化法制备姜黄素纳米胶束滴眼液,分别测定其粒径、包封率和稳定性;通过眼局部刺激实验方法考察纳米胶束的细胞毒性和细胞摄取作用;采用眼局部刺激实验方法测定姜黄素纳米胶束滴眼液的体内眼局部刺激性;以兔眼表给予花生四烯酸溶液建立眼局部炎症模型,评价纳米胶束的抗炎活性。结果使用聚己内酰胺-聚乙酸乙烯酯-聚乙二醇聚合物成功构建姜黄素纳米胶束滴眼液,粒径为(50.1±1.0)nm,包封率为(99.37±0.76)%;储存稳定性分析结果显示,12周后储存在4℃质量比为18∶1的姜黄素纳米胶束滴眼液药物含量仍有(98.13±0.97)%,而质量比15∶1的药物含量为(89.32±1.59)%。25℃条件下的实验结果与4℃结果相似,因此最佳处方质量比为18∶1。制备的姜黄素纳米胶束呈橙红色透明状,优化处方后的纳米胶束粒径、多分散系数和Zeta电位分别为(50.1±1.0)nm、0.079±0.011和-(1.83±0.67)m V,包封率为(99.37±0.76)%。姜黄素溶液在p H6.0至p H7.4的不同条件下,降解速度分别是对应姜黄素纳米胶束降解速度的24.9倍、28.4倍、44.8倍、73.7倍、315.0倍和662.1倍。体外细胞实验结果显示浓度为500.00μg·m L^(-1)的姜黄素纳米胶束孵育细胞48 h后,细胞存活率为86.89%;4.5 mg·m L^(-1)姜黄素纳米胶束滴眼液孵育细胞1 h后未发现细胞毒性。兔眼局部刺激性实验结果显示,姜黄素纳米胶束滴眼液、玻璃酸钠滴眼液(1 mg·m L^(-1))和空白PBS 3组实验在不同时间点的临床评分均在0~2的范围内,属于无刺激性等级。体内角膜渗透性结果显示,姜黄素纳米胶束组的荧光强度均显著强于姜黄素溶液组。抗炎活性结果显示姜黄素纳米胶束表现出显著的剂量依赖性抗炎活性。结论局部聚合物构建的姜黄素纳米胶束滴眼液显著改善了姜黄素的Objective To prepare a novel nanomicelle curcumin ophthalmic so- lution and investigate its physicochemical properties as well as biology properties in cells/animals level. Methods Nanomicelle curcumin ophthalmic solution was pre- pared by a solvent evaporation-hydration method,and its mean diameter, encapsation efficiency and stability were detected, respectively. The cytotoxicity and cellular uptake of the nanomicelle were tested by cellular experiments in vitro. The ocular irritation tests in vivo were investigated by using eye comprehensive score. The eye local inflam- matory models were set up by instilling arachidonic acid solution in rabbit eyes to evalu- ate the anti-inflammatory activity of nanomicelle. Results Nanomicelle curcumin oph- thalmic solution were successfully prepared by using polyvinyl caprolactam-polyvinyl ace- tate-polyethylene glycol graft copolymer （PVCL-PVA-PEG） with the mean diameter and encapsulation efficiency of （50.1± 1.0） nm and （99.37 ± 0.75） %. The remaining drugs of nanomicelle curcumin with 18 ： 1 and 15 ： 1 mass ratio were （98. 13±0.97）% and （89.32 ± 1.59）% after 12 weeks. The experimental results under 25 ℃ were similar to those under 4 ℃ ,so the best mass ratio was 18 ： 1. The curcumin nanomicelle was orange red and transparent,the nanomicelle particle size,polydispersity index and Zeta potential were （50. 1 ± 1.0） nm,0. 079 ± 0.011 and - （ 1.83± 0.57 ） mV, and the encapsulation effi- ciency was （99.37 ±0.75）%. Under different conditions of pH 6.0 to pH 7.4 ,the degra- dation rate of curcumin solution was 24.9,28.4,44.8,73.7,315.0,652.1 times respec- tively, which correspond to the degradation rate of curcumin nanoparticles. The cellular experiments in vitro showed that the cell viability were 85. 89% as the concentration of 500 μg mL-1 after cell incubation of 48 hours, and there was no cytotoxicity after cell incubation of 1 hour with 4.5 mg mL-1 nanomicelle curcumin. To determine the ocular tolerance in rabbits＇ eyes

关 键 词：姜黄素 聚己内酰胺-聚乙酸乙烯酯-聚乙二醇 纳米胶束 稳定性 抗炎活性 

分 类 号：R917[医药卫生—药物分析学] R77[医药卫生—药学]