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目的:优化仙鹤草多糖的提取工艺并测定其对脑胶质瘤U251细胞生长的抑制作用。方法:采用正交设计实验方法对影响仙鹤草多糖得率的提取温度、提取时间、料液比等3因素进行优化;采用MTT比色法测定仙鹤草多糖对U251细胞株增殖的抑制率,应用倒置相差显微镜观察仙鹤草多糖对肿瘤细胞形态结构的影响。结果:仙鹤草多糖提取的最佳工艺条件为:提取温度90℃,提取时间3 h,料液比1∶20,在此条件下,仙鹤草总多糖得率可达1.85%。仙鹤草总多糖对脑胶质瘤U251细胞株增殖具有明显的体外抑制作用(P<0.01),且表现出浓度依赖关系,IC50为1.35 g.L-1。经仙鹤草多糖处理后,U251细胞显示典型的凋亡形态变化。结论:优选的提取工艺简便,高效;仙鹤草总多糖的抗肿瘤效果显著,其抗肿瘤机制可能与诱导肿瘤细胞凋亡有关。
OBJECTIVE: To optimize the extraction process of Agrimony Agrimonia polysaccharides and determine its inhibitory effect on the growth of glioma U251 cells. Methods: Orthogonal design method was used to optimize the extraction temperature, extraction time and the ratio of solid to liquid of Agrimoner. The MTT assay was used to determine the inhibitory rate of Agrimony polysaccharide on the proliferation of U251 cells. Inverted phase contrast microscope observation Agrimonia polysaccharides on the morphology of tumor cells. Results: The optimal extraction conditions of Agrimoner were: extraction temperature 90 ℃, extraction time 3 h, ratio of material to liquid 1:20. Under these conditions, the yield of total agar could reach 1.85%. Agrimonia glyptostroboides significantly inhibited the proliferation of glioma U251 cells in vitro (P <0.01), and showed a concentration-dependent relationship with IC50 of 1.35 g · L -1. After Agrimony polysaccharide treatment, U251 cells showed a typical morphological changes of apoptosis. Conclusion: The optimal extraction process is simple and efficient. The antitumor effect of Agrimonia crassipes polysaccharide is significant, and its anti-tumor mechanism may be related to the induction of apoptosis of tumor cells.