[关键词]
[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(butyrylgalactose 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HepG2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HepG2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)mV。MTT实验表明,But-Gal-CMEs与CMEs对HepG2细胞增殖的半数抑制浓度(IC50)分别为62.55、71.23μg/mL。HepG2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HepG2对CMEs的细胞摄取,并增强其对HepG2细胞毒作用。
[Key word]
[Abstract]
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 71.23μg/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.
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[基金项目]
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)