灰树花多糖的分离纯化及体外抗肝癌作用研究  

Isolation,Purification and in Vitro Antitumor Activity of Polysaccharides GFD-1 from Grifola frondosa

作　　者：张媛媛[1] 孟梦[1] 王明飞[1] 王春玲[1] 

机构地区：[1]天津科技大学食品工程与生物技术学院教育部食品营养与安全重点实验室,天津300457

出　　处：《食品研究与开发》2016年第15期1-5,共5页Food Research and Development

基　　金："十二五"国家科技支撑计划(2012BADB33B04);国家自然科学基金(31000768)

摘　　要：利用超声破碎、水提醇沉、葡聚糖凝胶过滤层析等方法,对灰树花子实体多糖进行分离得到多糖组分GFD-1。高效液相色谱分析表明,GFD-1是分子量为29.574 k Da的多糖均一组分。利用显色试验、紫外光谱、红外光谱及原子力显微镜分析,GFD-1为不含还原性羰基、酚羟基、游离或结合蛋白质、核酸类物质的非淀粉类α-吡喃多糖,其糖链高度在0.5 nm^1 nm。GFD-1能够显著抑制人肝癌细胞HepG2的增殖,并呈剂量依赖关系,IC_(50)为94μg/m L,但其对于正常肝脏细胞L-02无明显影响;通过AO/EB双染,经GFD-1作用的HepG2细胞呈典型凋亡形态学特征;经流式细胞仪分析,HepG2细胞被阻遏在S期。上述结果可知,初步确定分离纯化的GFD-1是一种具有抗肿瘤活性的均一多糖组分,为开发天然抗肿瘤物质和进一步分析其抗肿瘤作用机制提供了参考。A polysaccharide GFD-1 from Grifolafrondosa fruiting bodies had been purified by sonication, water extraction, alcohol precipitation and Sephadex gel filtration chromatograp. By HPLC, GFD-1 was a kind of polysaccharide homogeneous components and its molecular weight was 29.574 kDa. The results of the color experiment, ultraviolet spectroscopy, infrared spectroscopy and atomic force microscope analysis illustrated that GFD-1 was a kind of non-starch a-pyran polysaccharide without reductive carbonyl, phenolic hydroxyl, free or a combination protein and nucleic acid; its sugar chain height was 0.5 nm-1 nm. GFD-1 inhibited proliferation of HepG2 cells in a dose-dependent manner, the ICs0 of GFD -1 was 94 μg/mL; it had no significant effect on the cell viability of humor normal liver L-02 cells. Meanwhile, in the presence of with GFD-1, HepG2 cells showed typical apoptotic morphological features through the AO/EB double staining. Flow cytometry analysis suggested that HepG2 cells were repressed at the phase S by GFD-1. These results showed that GFD-1 purified from Grifolafrondosa possesses anti-tumor regulation effect, serving a reference basis to develop natural anticancer drugs and further analyse the anti-tumor mechanism of GFD-1.

关 键 词：灰树花 多糖 分离纯化 抗肿瘤 

分 类 号：R284[医药卫生—中药学] R285[医药卫生—中医学]