天然产物研究与开发 ›› 2020, Vol. 32 ›› Issue (增刊1): 1-6.doi: 10.16333/j.1001-6880.2020.S.001

• 研究论文 •    下一篇

虫草素分离鉴定及对Hep G2细胞抑制研究

张虎成1，杨军1，范海涛1，王维彬1，杜天宇1，李泽亚1，毕磊1，尹逸奇1，胡宇1，王丁瑶1，张金军2，陈小峰2，张征田3*


  

  1. 1北京电子科技职业学院生物工程学院，北京 100170；2北京第35中学，北京 100020；3南阳师范学院生命科学与技术学院，南阳 473061 

  • 出版日期:2020-06-28 发布日期:2020-07-20
  • 基金资助:
    北京市教师队伍建设-职业院校素质提高工程（PXM201 8_014306_000057/2） 

Isolation and identification of cordycepin and its inhibitory effect on Hep G2 cells

ZHANG Hu-cheng1,YANG Jun1,FAN Hai-tao1,WANG Wei-bin1,DU Tian-yu1,LI Ze-ya1,BI Lei1,YIN Yi-qi1,Hu Yu1,WANG Ding-yao1,ZHANG Jin-jun2,CHEN Xiao-feng2,ZHANG Zheng-tian3*   

  1. 1Bioengineering College,Beijing Polytechnic,Beijing 100170,China;2Beijing No.35 High School,Beijing 100020,China;3Department of Life Sciences,Nanyang Normal University,Nanyang 473061,China 

  • Online:2020-06-28 Published:2020-07-20

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摘要： 从蛹虫草小麦培养基中提取和纯化虫草素，并进行了体外肝癌Hep G2细胞的抑制作用研究。以蛹虫草小麦培养基为原料，研究热超声水提取，探究pH值、温度、浸提时间、超声功率对虫草素的提取效果，用AB-8柱层析分离、重结晶纯化虫草素，并用HPLC、红外光谱鉴定。虫草素处理Hep G2肝癌细胞后,倒置相差显微镜下观察肝癌细胞形态变化,并用四甲基偶氮唑蓝（MTT）法测定细胞生长情况。结果表明，热超声水提取工艺为：固液比1∶50、pH值6、温度60 ℃、浸提时间3 h、超声功率300 W时提取效果好。经鉴定，分离提取物为虫草素，纯度达到99%以上。不同浓度的虫草素对Hep G2肝癌细胞有明显的抑制作用，且以浓度和时间呈正相关，被抑制的细胞变圆、凝集、碎裂增加。


关键词: 培养残基, 虫草素, 分离提取, 肝癌 G2细胞, 抑制

Abstract:

Cordycepin was extracted and purified from wheat medium of Cordyceps militaris and its inhibitory effect on Hep G2 cells was studied in vitro.The extraction of cordycepin was optimized using a thermosonic water bath at varying pH values, temperatures, extraction times and ultrasonic power.The results show that the extraction was better when solid to liquid ratio of 1∶50,at pH 6,at of 60 ℃,with an extraction duration of 3 hours,and 300 W ultrasonic power.The purity of cordycepin was over 99%.The extracted cordycepin was then purified employing a macroporous adsorptive resin (AB-8),and was identified by HPLC and Infrared spectroscopy.The growth of Hep G2 cells treated with the purified cordycepin was measured using the MTT assay,and the morphological changes of Hep G2 cells were observed by means of inverted differential microscopy.Different concentrations of cordycepin showed significant inhibitory effect on Hep G2 cells,and showed a positive correlation with concentration and inhibitory time.The inhibited cells became round,agglutinated and fragmented.

Key words: residual medium, cordycepin, extraction, Hep G2 cells, inhibitory effect ,

中图分类号:  Q936

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