摘要
目的:在细胞水平下,通过抗病毒实验,筛选佩兰最佳提取方法及敏感病毒株,并确定分离纯化效果最佳的大孔吸附树脂型号,为进一步分离非挥发油类抗病毒成分奠定基础.方法:先通过水提法、水提醇沉法、醇提法、醇提水沉法、乙酸乙酯提取法获得粗提物,在细胞水平下,由显微镜观察细胞病变效应(CPE),噻唑蓝(MTT)法染色,酶标仪测定吸光度值,通过比较治疗指数(TI),对肠道EV-71病毒、单纯疱疹Ⅰ型病毒、柯萨奇病毒B5、呼吸道合胞病毒进行体外抗病毒筛选;其次通过大孔吸附树脂分离法,以蒸馏水、体积分数为25%、50%、75%的乙醇作为洗脱剂冲洗树脂柱,筛选分离纯化效果最好的大孔吸附树脂.结果:经初步筛选,水提醇沉法中的沉淀对肠道EV-71病的抑制作用最强,TI为84.631;通过大孔吸附树脂法得,AB-8弱极性大孔吸附树脂对佩兰的分离纯化效果最好,其中体积分数为25%乙醇洗脱部位的T1值可达127.001.结论:体积分数为25%乙醇洗脱部位是佩兰抑制肠道EV-71病毒的最佳有效部位.
Objective： To screen for the best extraction method and the sensitive virus strain to Eupatorium Fortune Tuecz, determine the model with the best macroporous adsorption resin for separation and purification, and at the cellular level, to be achieved by antiviral experiments. In order to separate non-volatile oil antiviral components, establishment of the greatest solution for it was done. Methods： The crude extract was obtained by water extraction, water extraction and alcohol precipitation, alcohol extraction, alcohol extraction and water precipitation, and ethyl acetate extraction, sequentially. At the cellular level, the cytopathic effect （CPE） was observed by microscopy. The antiviral screening was performed on Human Enterovirus 71, Herpes Simplex Virus I , coxsackievirus B5 and Respiratory Syncytial Virus. Treatment index （TI） was an indicator to assess the inhibitory effect. And TI was obtained via thiazole blue （MTT） staining and the absorbance values was measured by microplatereader. The resin column was eluted using distilled water, 25% , 50% and 75% ethanol to screen the fittest solution for macroporous adsorption resin. Results： After preliminary screening, the TI of the precipitation imposing on EV-71 was much higher than others. Furthermore, AB-8 weak polarity macroporous adsorption resin was the best for separation and purification of Eupatorium Fortune Turcz, and TI at 25% ethanol elution was up to 127.001. Conclusion：25% ethanol elution is the best effective part of Peynland which inhibits intestinal Human Enterovirus 71.
作者
刘飞
徐佳馨
颜娓娓
刘文静
周长征
LIU Fei;XU Jiaxin;YAN Weiwei;LIU Wenjing;ZHOU Changzheng(College of Pharmacy, Shandong University of Traditional Chinese Medicine, Jinan 250355, Chin)
出处
《暨南大学学报（自然科学与医学版）》
CAS
CSCD
北大核心
2018年第1期80-86,共7页
Journal of Jinan University(Natural Science & Medicine Edition)
基金
山东省中医药研究及产业化课题项目(XTCX2014C01-04)
关键词
佩兰
提取方法
大孔吸附树脂
体外抗病毒
Eupatorium Fortune Tuecz
extraction method
macroporous adsorption resin
vitroantiviral
