丝棉木果实多糖制备及其体外抑瘤活性

Preparation of polysaccharides from Euonymus bungeanus fruit and anti-tumor activity

【目的】研究丝棉木果实多糖的提取工艺,分析其体外抗小鼠骨髓瘤细胞活性,为丝棉木的开发利用提供依据。【方法】以丝棉木果实为原料,采用水提醇沉法提取多糖并用硫酸苯酚法测定多糖含量,以提取时间、提取温度、料(g)液(mL)比为影响因素,以多糖得率为考察指标进行单因素试验,在此基础上,采用响应面法对提取工艺条件进行优化,并采用结晶紫法检测丝棉木果实多糖体外抗小鼠骨髓瘤活性的IC_(50)。【结果】丝棉木果实多糖的水浴提取最佳工艺为:料液比1∶30,温度90℃,提取时间1.5h,得率为10.90%,经纯化获得纯度达90.23%的多糖。光学显微镜下观察到,丝棉木果实多糖处理组小鼠骨髓瘤细胞SP2/0出现典型的细胞凋亡形态改变。丝棉木果实多糖对小鼠骨髓瘤细胞SP2/0有明显的抑制作用,IC_(50)为37.753μg/mL。【结论】确定了丝棉木果实多糖的最佳提取工艺,该多糖对小鼠骨髓瘤细胞SP2/0有明显的抑制作用。

[Objective] This study optimized the extracting technology of polysaccharides from Euonymus bungeanus fruits and investigated the in vitro anti tumor activity against mouse myeloma cells to provide basis for the development and utilization of E. bungeanus. [Method] The polysaccharides were extrac ted ftom fruits of E. bungeanus by water extraction and alcohol precipitation methods, and the content of polysaccharides was determined by phenol sulfate method. Single factor experiments were preformed using extraction time, extraction temperature and solid liquid ratio as influential factors, and the response surface software was used to optimize the extract conditions based on polysaccharides yield. IC50 of anti tumor ac tivity of the polysaccharides against mouse myeloma ceils was determined by crystal violet method. [Result] The best conditions for the extract process of polysaccharides were ratio of solid to liquid 1 ：30,wa ter bath temperature 90℃, and time of extraction 1. 5 h. The yield under the optimal conditions was 10.90% ,and the purity was 90. 23% . The apoptosis morphological changes of the myeloma cells SP2/0 were observed under photon microscope after the treatment with polysaccharides of E. bungeanus fruits.The activity tests showed that the IC50 was 37. 753 μg/mL. [Conclusion] This study obtained the optimal extraction conditions of polysaccharides from fruits of E. bungeanus and the extracted polysaccharides had inhibitory effect on mouse myeloma cells SP2/O.