丁酰半乳糖酯修饰的薏苡仁组分微乳的制备及其体外抗肿瘤活性研究  

Study on preparation of butyryl galactose ester-modified coix component microemulsions and their anticancer activity in vitro

作　　者：刘明健[1,2] 瞿鼎[2] 陈彦[2] 孙文杰[2] 袁成甜 王理想[2] 

机构地区：[1]江苏大学药学院,江苏镇江212013 [2]江苏省中医药研究院中药组分与微生态研究中心,江苏南京210028

出　　处：《中草药》2015年第18期2696-2702,共7页Chinese Traditional and Herbal Drugs

基　　金：国家自然基金资助项目(81373979);江苏省自然科学基金资助项目(BK20141508)

摘　　要：目的合成丁酰半乳糖酯(butyryl galactose ester,But-Gal)并制备丁酰半乳糖酯修饰的薏苡仁组分微乳(butyryl galactose ester modified coix component microemulsions,But-Gal-CMEs),对其进行理化性质评价和体外抗肿瘤活性测定。方法采用酶催化法合成But-Gal,并通过1H-NMR与FT-IR对其结构进行表征。以薏苡仁油为油相,聚氧乙烯氢化蓖麻油(Cremophor®RH40)为乳化剂,PEG400为助乳化剂,But-Gal为肝肿瘤细胞靶配体,薏苡仁多糖水溶液为水相,水滴定法制备微乳,测定其粒径、电位和形态。通过MTT法考察薏苡仁组分微乳(CMEs)与But-Gal-CMEs对肿瘤细胞Hep G2细胞毒作用;以异硫氰酸荧光素(FITC)为荧光探针,借助荧光倒置显微镜观察Hep G2细胞对微乳的摄取行为。结果经1H-NMR与FT-IR表征确认丁酰半乳糖酯结构与设计一致。制备所得But-Gal-CMEs外观形态圆整,平均粒径为(57.68±6.65)nm,多分散指数(PDI)为0.070±0.006,Zeta电位为(-2.95±0.23)m V。MTT实验表明,But-Gal-CMEs与CMEs对Hep G2细胞增殖的半数抑制浓度(IC50)分别为62.55、71.23μg/m L。Hep G2细胞摄取结果显示But-Gal-CMEs组的荧光强度强于CMEs组。结论所制备的But-Gal-CMEs粒径小,外观圆整,稳定性好,But-Gal能增加Hep G2对CMEs的细胞摄取,并增强其对Hep G2细胞毒作用。Objective To synthesize butyryl galactose ester（But-Gal） and prepare butyryl galactose ester-modified coix component microemulsions（But-Gal-CMEs） and to evaluate its physicochemical properties and anticancer activity in vitro.Methods But-Gal was synthesized by enzyme-catalyzed reaction and the structure of the product was confirmed by ～1H-NMR and FT-IR.The CMEs and But-Gal-CMEs were prepared by aqueous titration method using coix seed oil,Cremophor RH40,PEG400,But-Gal,and coixan solution as oil phase,surfactant,cosurfactant,target ligand,and aqueous phase,respectively.The average particle size,polydispersity index（PDI）,and Zeta potential were detected by dynamic light scattering（DLS）.The cytotoxicity of CMEs and But-Gal-CMEs aganist HepG2 cells was determined by MTT assay.The cellular uptake of CMEs and But-Gal-CMEs was detected by fluorescence microscopy.Results The structure of But-Gal was confirmed by ^1H-NMR and FT-IR.The But-Gal-CMEs displayed the spherical surface with mean droplet size of（57.68±6.65） nm,PDI of 0.070±0.006,and Zeta potential of（-2.95±0.23） mV,respectively.MTT experiments showed that the half of HepG2 cell proliferation inhibition concentration（IC50） of But-Gal-CMEs and CMEs was 62.55 and 71.23 μg/mL.The HepG2 cell uptake results suggested that the fluorescence intensity of But-Gal-CMEs group was higher than that of CMEs group.Conclusion The But-Gal-CMEs presents small particle size,good roundness,and good stability.In addition,But-Gal could increase the uptake rate of CMEs in HepG2 cells and enhance the inhibition of HepG2 cell proliferation.

关 键 词：薏苡仁 丁酰半乳糖酯 微乳 细胞毒性 细胞摄取 抗肿瘤活性 酶催化法 水滴定法 异硫氰酸荧光素 

分 类 号：R283.6[医药卫生—中药学]