知母皂苷B-Ⅱ纳米纤维膜制备及体外抗肿瘤活性的初步研究

Preparation of timosaponin B-Ⅱ nanofiber membranes and preliminary study on its antitumor activity in vitro

目的:探索知母皂苷B-Ⅱ纳米纤维膜的制备方法,为后续进行体内相关研究打好基础。方法:将知母皂苷B-Ⅱ与聚乳酸(PLLA)按照不同比例制成知母皂苷B-Ⅱ纳米纤维膜;分别采用扫描电镜、红外光谱分析、LC-MS/MS检测方法对纤维膜理化性质表征、生物相容性、缓释性能等进行测试;采用CCK-8法检测载药纳米膜缓释药物抑制胃癌细胞的增殖情况。结果:12%的知母皂苷B-Ⅱ浓度为最佳载药浓度;在制备载知母皂苷B-Ⅱ纳米纤维的过程中,静电纺丝技术能保持两种组分各自的化学特征及物理性质;知母皂苷B-Ⅱ对纤维膜的热稳定性影响不大,两者相容性较好;知母皂苷B-Ⅱ纳米纤维膜可有效释放知母皂苷B-Ⅱ,明显抑制BGC-823细胞的增殖活性,在48 h、72 h与对照组细胞相比较,差异具有统计学意义(P<0.05)。结论:知母皂苷B-Ⅱ纳米纤维膜通过缓释药物可有效抑制BGC-823细胞的增殖能力。

Objective： To explore the preparation method of timosaponin B-Ⅱ nanofiber membrane, in order to lay afoundation for the subsequent studies in vivo. Methods： Timosaponin B-Ⅱ and PLLA at different ratios were made into timosaponinnanofiber membranes; the surface structure and morphology of the drug loaded nanofiber membranes were observed by scanningelectron microscopy; the infrared spectra was adopted to determine the compatibility between the drug and the polymer; LC-MS/MSwas used to measure timosaponin B-Ⅱ levels in solutions at different time points to draw the drug release curves; CCK-8 assay wasused to detect the proliferative activity of tumor cells, in order to evaluate whether the sustained drug release from the drug-loadednano-films could inhibit the proliferation of gastric carcinoma cells. Results： The optimum concentration of timosaponin B-Ⅱ fordrug loading was 12%; the electrospinning technique in the preparation of timosaponin B-Ⅱ loaded nanofibers could maintainthe physical and chemical properties of the both components; timosaponin B-Ⅱ had little effect on the thermal stability of fibermembranes, indicating a good compatibility; the timosaponin B-Ⅱnanofiber membranes could effectively release timosaponin B-Ⅱinthe conventional culture solution, the timosaponin B-Ⅱloaded nanofiber membranes can inhibit the proliferation of BGC-823 cellssignificantly, the differences in the treated groups at 48 hours and 72 hours were significant （P 〈 0.05） compared with the controlgroup. Conclusion： Timosaponin B-Ⅱ nanofiber membranes can effectively inhibit the proliferation of BGC-823 cells through thesustained release of the drug.