阴地蕨多糖提取工艺及抗肿瘤活性测定的研究.docVIP

阴地蕨多糖提取工艺及抗肿瘤活性测定的研究 　　摘要 水提醇沉法提取阴地蕨多糖，以多糖得率为指标，考察提取温度、提取时间、料液比及提取次数对多糖得率的影响，通过正交试验优化提取工艺参数；采用MTT法检测阴地蕨多糖对癌细胞CT-26、K562、WEHI-3细胞作用48 h后的增殖抑制作用。试验结果表明，提取时间和温度是阴地蕨多糖提取率最主要的影响因素，优化的最佳提取条件为：温度100 ℃，料液比为1∶40（mg/mL），提取时间4 h和提取次数3次。阴地蕨多糖对3种肿瘤均有生长抑制作用，且呈现一定的浓度依赖性，抑制作用强弱为WEHI-3K562CT-26，半数抑制浓度IC50分别为0.85、2.13、2.52 mg/mL。 　　关键词 阴地蕨多糖；水提醇沉法；正交试验；MTT法；抗肿瘤活性 　　中图分类号 S567.239 文献标识码 A 文章编号 1007-5739（2016）15-0261-02 　　Abstract The Botrychium ternatum（Thunb） Sw. polysaccharides was extracted by water extraction-alcohol precipitation method. The extraction process was optimized by investigating polysaccharide yield as the response value of extraction time，extraction temperature，solid to solvent ratio and extraction time. Based on the results of single-factor experiments，orthogonal array experiments were carried out to determine the optimum process parameters. MTT assay was adopted to detect the inhabitation of polysaccharides with three different kinds of malignant tumor cell lines in vitro and IC50 values were calculated，respectively.The results showed that the best conditions for water extraction-alcohol precipitation were determined as follows：extraction temperature was 100 ℃；the time was 4 h；solid to liquid ratio was 1∶40（g/mL）；and the extractiontime was 3 times. The polysaccharides had antitumor effects on WEHI-3K562CT-26，they inhibited cell proliferation in a dose dependent manner，and the IC50 of polysaccharides on the three different kinds cell lines was 0.85，2.13，2.52 mg /mL，respectively. 　　Key words Botrychium ternatum（Thunb）Sw.polysaccharide；water extraction-alcohol precipitation；orthogonal array experiments；MTT assay；antitumor activity 　　阴地蕨[Botrychium ternatum（Thunb）Sw.]属于阴地蕨科阴地蕨属植物，阴地蕨属植物全世界有40余种，主要分布于温带的高山地区和低地森林地区，我国现有17种，分布于全国各地。该属植物具有清热解毒、利水通淋、消肿等广泛的药理活性[1]，具有很高的药用价值，民间多用阴地蕨属植物治疗肺热咳嗽、毒蛇咬伤、出血等疾病症状而受到了人们的关注[2]。其中阴地蕨，别名一朵云、小春花、四块瓦等[3]，其味淡、苦，性冷，有止咳、清肺热之功效[4]；王少明等[5]从阴地蕨中提取的木犀草素、阴地蕨素等提取物具有抗肿瘤转移作用，具体表现为抑制自发性肿瘤转移和实验性肿瘤转移；陈晓清等[6]从阴地蕨分离得到的粗多糖提取物具有抑制鱼病病原菌包括烂尾病病原菌、赤皮病病原菌