矢车菊素3-葡萄糖苷膜修饰脂质体的制备及其对HLECs保护作用的初步考察

Preparation of Membrane-modified Liposomes Loaded with Cyanidin-3-glycoside and Its Primary Protective Effect on HLECs

采用逆相蒸发法制备矢车菊素3-葡萄糖苷(1)脂质体(1-L)。利用星点设计-效应面法,以包封率为评价指标,优化得到的处方为:1浓度7.0 mg/ml、磷脂与胆固醇质量比4、有机相与水相体积比4、磷酸盐缓冲液浓度15 mmol/L。再用三甲基壳聚糖对1-L进行膜修饰,所得标题脂质体(1-TCL)的粒径为(158.3±2.8)nm,?电位为(31.7±1.0)m V,包封率为(53.7±0.2)%,在人工泪液中36 h累积释放率约40%。以人晶体上皮细胞(HLECs)为模型,考察了1溶液、1-L和1-TCL对双氧水致氧化损伤HLECs进行先预处理再治疗的保护作用。与模型组细胞相比,40和80?mol/L的1-TCL组细胞的活力提高了64.7%与74.4%。与1-L(40?mol/L)组相比,1-TCL组对HLECs细胞的保护作用均显著提高(P<0.05)。

The cyanidin-3-glycoside （1） liposomes （1-L） were prepared by reverse-phase evaporation. The formulation was optimized by a central composite design-response surface method with entrapment efficiency as index. The results showed that the optimal parameters of 1-L were as follows： 1 concentration was 7.0 mg/ml, the weight ratio of phospholipid to cholesterol was 4, the volume ratio of organic phase to water phase was 4, and the concentration of phosphate buffer was 15 mmol/L. Then the optimal 1-L was coated with N-trimethyl chitosan （TMC） to obtain the title liposomes （1-TCL）. The particle size, ζ potential and entrapment efficiency were （158.3±2.8）nm, （31.7±1.0）mV and （53.7±0.24） %, respectively. In vitro cumulative release at 36 h was about 40% in artificial tear fluid. The human lens epithelium cells （HLECs） were pretreated by 1 solution, 1-L or 1-TCL, followed by oxidative damage with hydrogen peroxide and treated by corresponding samples again. Then the protective effects of 1 solution, 1-L and 1-TCL were investigated through above cell models. In comparison with the model group damaged by hydrogen peroxide, the cell viabilities were elevated by 64.7% and 74.4% in 1-TCL groups at 40 and 80 μmol/L. Compared with 1-L （40 μmol/L）, more significant protective effects of 1-TCL were exhibited （P〈0.05）.