仙鹤草多糖的提取及其体外抗脑胶质瘤U251活性研究

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3h,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.

Extraction of Total Polysaccharides from Agrimonia pilosa and Evaluation of its Inhibitory Activity on Human Glioblastoma U251 Cells in vitro

Objective:The optimum extraction technology of total polysaccharides from Agrimonia pilosa was established and its inhibitory activity on human glioblastoma U251 cells in vitro was evaluated.Method: The orthogonal test was implied to optimize the extraction process conditions of total polysaccharides.With the extraction rate as the index,the extraction temperature,extraction time and liquid-material ratio were optimized.MTT assay was used to determine the effect of total polysaccharides on cell growth inhibitory.Morphological changes of the U251 cells were observed through inverted phase contrast microscope.Result: The optimum extraction conditions were determined as following: extraction temperature 90 ℃,extraction time 3 hours and liquid-material ratio of 1∶20.Under these conditions the maximal yield of total polysaccharides reached 1.85%.Total polysaccharides from A.pilosa could inhibit the hyperplasia of glioblastoma U251 cells in dose-dependent manner(P<0.01) and its IC50 reached 1.35 g·L-1.After treatment with polysaccharides U251 cells showed apoptosis morphology.Conclusion: The optimized extraction technology is effective and simple.Total polysaccharides from A.pilosa significantly inhibited tumor cell growth in vitro by inducing apoptosis of tumor cells.