蛹虫草培养基多糖的提取及抗肿瘤活性研究

Research on Extraction and Antineoplasmic Activity of Polysaccharide in Culture Medium ofCordyceps militaris

DOI：10.3969/j.issn.1673-1689.2019.04.018

中文关键词: 蛹虫草 培养基 多糖 抗肿瘤

英文关键词: Cordyceps militaris,culture medium,polysaccharide,antineoplastic

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中文摘要:

为综合利用蛹虫草小麦培养基，研究培养基多糖的提取和开发。在热回流水提法和超声波水提法单因素试验的基础上，采用热回流水提法正交试验筛选蛹虫草小麦培养基粗多糖最佳的提取工艺；用 MTT法检测了不同处理提纯多糖的抗肿瘤活性。结果表明热回流水提法提取培养基粗多糖效果优于超声波水提法；提取温度100 ℃、料液比1 g∶25 mL、提取3次、每次提取时间90 min，为最佳培养基粗多糖提取工艺；影响粗多糖提取效率的4个因素的主次关系是：提取温度>提取时间>提取次数>料液比；粗多糖的得率随着醇沉浓度的提高而提高。MTT试验发现质量浓度为5 mg/mL、培养时间72 h，培养基脱脂脱蛋白多糖A对HepG2肝癌细胞抑制率可达到83.02%，培养基脱脂多糖B的抑制率可达到75.39%，培养基未脱脂脱蛋白粗多糖C抑制率为30.61%，提纯多糖表现出较高的抑制率；100 ℃热回流水提蛹虫草子实体脱脂脱蛋白多糖F，质量浓度为5 mg/mL、培养72 h后，对Hepg2细胞抑制率平均值达到92.974%，与对照顺铂（SB）无显著差异，与多糖A差异显著。

英文摘要:

The extraction and development of polysaccharide was researched to multipurpose use of cordyceps militaris culture medium with wheat. This article studied the optimum extraction process of coarse polysaccharide in cordyceps militaris culture medium through the orthogonal factor experiment on hot reflux water on the base of single factor experiment on hot reflux water and ultrasonic extraction，researched the antitumor activity of polysaccharide purified by cordyceps militaris culture medium by MTT method. The results showed that hydrothermal refluxing extraction is better than supersonic water extraction on the basis of the results of single factor experiment. The results of orthogonal test showed the optimal technological conditions for the extraction conditions of coarse polysaccharide of were as follow：extracting temperature was 100 ℃， material-water ratio was 1∶25，extracting times was three and time was 90 min. The primary and secondary relationships of four Factors influencing the efficiency of extraction coarse polysaccharide are：extraction temperature > extraction time > extraction times > solid-liquid ratio. The coarse polysaccharide yield increased with the increasing of concentration of alcohol. The MTT test found the polysaccharide purified by different processing has inhibitory effect on the liver cancer cells，5 mg/mL concentration of polysaccharide，effect time for 72 h，inhibition rate of polysaccharide gained by degreasing and deproteinization from culture medium of Cordyceps militaris for hepg2 cells can reach 83.02%，inhibition rate of polysaccharide gained by degreasing medium for hepg2 cells can reach 75.39%，inhibition rate of polysaccharide gained without degreasing and deproteinization for hepg2 cells can reach 30.61%，purified polysaccharide showed higher inhibition rate. The polysaccharide extracted by cordyceps militaris fruiting body with 100 ℃ hot reflux water ，5 mg/mL concentration of polysaccharide，training after 72 hours，the inhibition rate reached 92.974% on Hepg2 cell，there was no significant difference compared with SB，but was significant difference compared with polysaccharide gained by degreasing and deproteinization from culture medium ofCordyceps militaris.

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