中草药2026，Vol.57Issue(17)：6715-6729,15.DOI:10.7501/j.issn.0253-2670.2026.17.006
Box-Behnken设计-效应面法优化氯化两面针碱脂质体处方工艺及体外抗胆管癌活性研究
Study on formulation optimization of nitidine chloride liposomes by Box-Behnken design-response surface method and anti-cholangiocarcinoma activity in vitro
摘要
Abstract
Objective To optimize the preparation process of nitidine chloride liposomes(NC-Lips),characterize their physicochemical properties,and evaluate their in vitro anti-cholangiocarcinoma activity.Methods NC-Lips were prepared by the thin-film dispersion method.Three key formulation variables,namely DSPE-mPEG2000 dosage,drug-to-lipid ratio,and phospholipid-to-cholesterol ratio,were selected.The formulation was optimized via single-factor experiments combined with Box-Behnken design-response surface methodology(BBD-RSM),with encapsulation efficiency,drug loading,PDI,and ζpotential as the evaluation indices.The drug content was determined by HPLC.The physicochemical properties of NC-Lips were characterized using a particle size analyzer and transmission electron microscopy.The in vitro drug release profile was investigated by the dialysis bag method.CCK-8 assay,flow cytometry,and cell cycle analysis were employed to evaluate the proliferation inhibition,apoptosis induction,and cell cycle arrest effects of NC-Lips on cholangiocarcinoma TFK-1 cells.Results The optimal formulation was as follows:DSPE-mPEG2000 dosage of 34.79 mg,drug-to-lipid ratio of 1∶15.15,and phospholipid-to-cholesterol ratio of 17.88.The prepared NC-Lips exhibited typical liposomal vesicular morphology,with an average particle size of(77.67±0.75)nm,PDI of 0.212±0.01,ζpotential of(-11.43±0.12)mV,encapsulation efficiency of(85.96±0.61)％,and drug loading of(4.37±0.03)％.Among the in vitro drug release models,the Ritger-Peppas model showed the highest goodness of fit,indicating that NC-Lips had obvious sustained-release characteristics.The liposomes remained stable during 30 d storage at 4 ℃,with a drug leakage percentage of＜30％.In vitro cell experiments demonstrated that NC-Lips exerted significantly better effects on proliferation inhibition,apoptosis induction,and G2/M phase arrest in TFK-1 cells than free nitidine chloride(P＜0.01).At a concentration of 12 μmol/L,the cell viability was only(8.17±1.90)％after 48 h of treatment;at the same concentration,the apoptosis rate reached(28.90±2.38)％at 24 h,and the proportion of cells in the G2/M phase increased to(66.23±3.31)％.Conclusion A stable and optimized NC-Lips formulation with excellent physicochemical properties was successfully prepared.NC-Lips exerted excellent in vitro anti-cholangiocarcinoma activity by inducing apoptosis and G2/M phase arrest.These findings provide an experimental basis for formulation optimization and subsequent mechanistic and in vivo studies of nitidine chloride.关键词
氯化两面针碱/Box-Behnken设计-效应面法/脂质体/胆管癌/细胞凋亡Key words
nitidine chloride/Box-Behnken design-response surface method/liposomes/cholangiocarcinoma/apoptosis分类
医药卫生引用本文复制引用
黄艳秋,丘炜桉,陈海燕,彭诗雅,苏明哲,刘悦,叶勇..Box-Behnken设计-效应面法优化氯化两面针碱脂质体处方工艺及体外抗胆管癌活性研究[J].中草药,2026,57(17):6715-6729,15.基金项目
广西自然科学基金资助项目(2022GXNSFDA035063) （2022GXNSFDA035063）
广西自然科学基金资助项目(2023GXNSFAA026366) （2023GXNSFAA026366）
国家自然科学基金面上项目(82473885) （82473885）
广西医科大学一流学科创新驱动人才计划 （）
